	   SnapGene      L_agtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgcgtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgagcggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagcatcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcagccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagact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!"\##$e%k%y'R)l*!+Q+--10@0w0z1B8DDGGG!HHIIK,KYK3L/  ~a;	?kq*defg!#k00y1A8?AG	HH*K+K2L  P0  GACNNNNNNGTC,CCAGGCCTGG,GATCCGGATC,CCWGGTACCWGG,CCANNNNNTGG,CCAGGNNNTGG,CCANNNCCTGG,CCAGGNCCTGG,GCACNNNAACGTTNNNGTGC,CACNNNGTG,CCNNNNNNNGG,AACNAACRYGTGCGTGC,GCACGCACRYGTTNGTT,GCACNAACRYGTTNGTGC,AACGCACRYGTGCGTT,GCACNNNNNNGTTSAACNNNNNNGTGC,GACNNNNNGTC,GAANNNNNNNTTGG,CCAANNNNNNNTTC,CACNNNNGTG,AACACNNNNGTGC,GCACNNNNGTGTT,GCANNNNNNTGC,GAACNNNNNNTCC,GGANNNNNNGTTC,CAGNNNCTG,CCAGGNNCTG,CAGNNCCTGG,CCWGGGCCC,GGGCCCWGG,CCWGGGCCCWGG,AACNNNNNNGTGCACNNNNNNGTT,GACNNNNNNTTYG,CRAANNNNNNGTC,GACNNNGTC,GAANNNNTTC,CCWGGTACC,GGTACCWGG,CCWGGWCCWGG,ACNNNNGTAYC,GRTACNNNNGT,CCWGGYRCCWGG,GAAGNNNNNNTAC,GTANNNNNNCTTC,AACGGCNNNGTGC,GCACNNNGCCGTT,GCACGGCNNNGTT,AACNNNGCCGTGC,CGANNNNNNTGC,GCANNNNNNTCG,CGATCNNNNTGC,GCANNNNGATCG,TGANNNNNNTCA,TGATCNNNNTCA,TGANNNNGATCA,TGATCNNGATCA,AACCTGCNNGTGC,GCACNNGCAGGTT,GCACCTGCNNGTT,AACNNGCAGGTGC,GCCNNNNNGGC,AACNNNNNNGTGCMC,GKGCACNNNNNNGTT,CCWGGNCCWGG,AACTGGGNNGTGC,GCACNNCCCAGTT,GCACTGGGNNGTT,AACNNCCCAGTGC,GACNNNNGTC,GAGNNNNNCTC,CCWGGTCTC,GAGACCWGG,GATNNNNATC,GATCNNNATC,GATNNNGATC,GATCNNGATC,CCWGGCGCCWGG,ACNNNNNCTCC,GGAGNNNNNGT,AACNNNNNNGTGCAG,CTGCACNNNNNNGTT,GATCATGATC,CCWGGNNCCWGG,AACTGGNNNGTGC,GCACNNNCCAGTT,GCACTGGNNNGTT,AACNNNCCAGTGC,AACNAACCGGTGCGTGC,GCACGCACCGGTTNGTT,GCACNAACCGGTTNGTGC,AACGCACCGGTGCGTT,GCANNNNNTGC,GCACNNNNNNGTTCGAACNNNNNNGTGC,CCTNNNNNAGG,GCNNNNNNNGC,CCANNNNNNTGG,CCAGGNNCCTGG,CCTGGCCAGG,CAANNNNNGTGG,CCACNNNNNTTG,TTTAAACNNNNNNGTGC,GCACNNNNNNGTTTAAA,GCACNNNNNNGTTTAAACNNNNNNGTGC,CCAGGNCCY,RGGNCCTGG,CCWGGGNCCTGG,CCAGGNCCCWGG,AAGNNNNNCTT,GAYNNNNNVTC,GABNNNNNRTC,GAYNNNNNRTC,GATCNNNNVTC,GABNNNNGATC,GAYNNNNGATC,GATCNNNNRTC,GATCNNNGATC,GTYAACNNNNNNGTGC,GCACNNNNNNGTTRAC,GCACNNNNNNGTTAACNNNNNNGTGC,GATCNGATC,TAACTATAACGGTCCTAAGGTAGCGA,TCGCTACCTTAGGACCGTTATAGTTA,CTCTCTTAAGGTAGC,GCTACCTTAAGAGAG,TAGGGATAACAGGGTAAT,ATTACCCTGTTATCCCTA,GTTAACNNNNNNGTGC,GCACNNNNNNGTTAAC,AACNAACGCGTGCGTGC,GCACGCACGCGTTNGTT,GCACNAACGCGTTNGTGC,AACGCACGCGTGCGTT,TCCAACNNNNNNGTGC,GCACNNNNNNGTTGGA,CAYNNNNRTG,AACAYNNNNGTGC,GCACNNNNRTGTT,GTTTAAACNNNNNNGTGC,GCACNNNNNNGTTTAAAC,CCWGGCGCC,GGCGCCWGG,CCWGGNNCC,GGNNCCWGG,GATCGCGATC,AACNAACATGTGCGTGC,GCACGCACATGTTNGTT,GCACNAACATGTTNGTGC,AACGCACATGTGCGTT,TGGCAAACAGCTATTATGGGTATTATGGGT,ACCCATAATACCCATAATAGCTGTTTGCCA,ATCTATGTCGGGTGCGGAGAAAGAGGTAATGAAATGG,CCATTTCATTACCTCTTTCTCCGCACCCGACATAGAT,AACNNGCACGTGCNNGTT,GAACNNNNNCTC,GAGNNNNNGTTC,CCAGGWCCY,RGGWCCTGG,CCWGGGWCCTGG,CCAGGWCCCWGG,CCAGGWCCTGG,TTATAACNNNNNNGTGC,GCACNNNNNNGTTATAA,GCACNNNNNNGTTATAACNNNNNNGTGC,GAACNNNNNNTAC,GTANNNNNNGTTC,GGCCNNNNNGGCC,AACNNACTAGTGC,GCACTAGTNNGTT,AACTAGTNNGTGC,GCACNNACTAGTT,AACNAACTAGTGCGTGC,GCACGCACTAGTTNGTT,GCACNAACTAGTTNGTGC,AACGCACTAGTGCGTT,AACNNNAACGTGCNNGTGC,GCACNNGCACGTTNNNGTT,TTAACNNNNNNGTGC,GCACNNNNNNGTTAA,CACNNNNNNTCC,GGANNNNNNGTG,CACNNNNGATCC,GGATCNNNNGTG,GATCTAGATC,CCANNNNNNNNNTGG,CCAGGCCT,AGGCCTGG,CCTCGAGG,GATCCGGA,TCCGGATC,GGCGCGCC,GCGATCGC,CCWGGWCC,GGWCCWGG,GATCGATC,GATCATGA,TCATGATC,GCGGCCGC,CCTGGCCR,YGGCCAGG,CCWGGNCC,GGNCCWGG,GGCCGGCC,RTGCGCAY,GATCNNGA,TCNNGATC,CGCGCGCG,CCTGGCCA,TGGCCAGG,CGCCGGCG,GTTTAAAC,GATCGCGA,TCGCGATC,TTAATTAA,VCTCGAGB,CCTGCAGG,CRCCGGYG,CGTCGACG,ATTTAAAT,GCCCGGGC,GATCTAGA,TCTAGATC,CACCTGC,GCAGGTG,CCTNAGG,GATCGAT,ATCGATC,CCTCAGC,GCTGAGG,GCTNAGC,CCTNAGC,GCTNAGG,GCTCTTC,GAAGAGC,GGTNACC,CGGWCCG,ACCWGGT,RGGNCCY,CCTGGAG,CTCCAGG,GGTGATC,GATCACC,GATCNGA,TCNGATC,GGGWCCC,GAAGATC,GATCTTC,CCCWGGG,TCCNGGA,TCCWGGA,RGGWCCY,TTATAA,AGGCCT,GACGTC,GTMKAC,TCCGGA,TGCGCA,ACCTGC,GCAGGT,GGTACC,AGTACT,GGYRCC,CCGCTC,GAGCGG,AACGTT,YGGCCR,RAATTY,CTGAAG,CTTCAG,CACGTG,GRCGYC,AGCGCT,CTTAAG,ACRYGT,ACCGGT,ACTAGT,CACGTC,GACGTG,GWGCWC,GTGCAC,CYCGRG,GGGCCC,ATTAAT,CCTAGG,TTCGAA,GCTAGC,GKGCMC,TGGCCA,GGATCC,GRGCYC,ATCGAT,CACGAG,CTCGTG,GGCGCC,GAAGAC,GTCTTC,GCATGC,GTATCC,GGATAC,TGATCA,ACTGGG,CCCAGT,CTRYAG,RGCGCY,AGATCT,GGNNCC,CTGGAG,CTCCAG,CTTGAG,CTCAAG,CGATCG,GGTCTC,GAGACC,YACGTR,CCNNGG,GAATGC,GCATTC,WCCGGW,GCAATG,CATTGC,RCCGGY,GCGCGC,GAGGAG,CTCCTC,CGGCCG,CCCAGC,GCTGGG,GTGCAG,CTGCAC,CGRYCG,CGTACG,CGTCTC,GAGACG,CCATGG,TCGCGA,GDGCHC,TGTACA,TCATGA,ACATGT,GTATAC,CCWWGG,CTCTTC,GAAGAG,GCNNGC,CCRYGG,GTTAAC,RCATGY,TACGTA,RGATCY,GCGATG,CATCGC,GCAGTG,CACTGC,CCCGGG,CCGCGG,TTTAAA,GGCGGA,TCCGCC,GAGCTC,GATATC,CTGRAG,CTYCAG,CAGCAG,CTGCTG,GAATTC,ATGCAT,CATATG,GTYRAC,AAGCTT,GTNNAC,TCNNGA,CAATTG,ACGCGT,TCCRAC,GTYGGA,GCCGGC,CMGCKG,GCCGAG,CTCGGC,CTCGAG,CTGCAG,CAGCTG,GTCGAC,CTYRAG,AATATT,GATCGA,TCGATC,GACCGA,TCGGTC,WGTACW,TARCCA,TGGYTA,TCTAGA,GGATC,GATCC,TTSAA,CCWGG,GTCTC,GAGAC,GCWGC,GGNCC,CCSGG,GGTGA,TCACC,GGWCC,GCAGC,GCTGC,CCATC,GATGG,ACGGC,GCCGT,GCNGC,CCNGG,GCATC,GATGC,ACTGG,CCAGT,GGATG,CATCC,CTCAG,CTGAG,GGGAC,GTCCC,ACNGT,CTNAG,CAGTG,CACTG,GACGC,GCGTC,CCCGC,GCGGG,GANTC,CGWCG,TCNGA,CCTTC,GAAGG,GCTCC,GGAGC,GTNAC,GAAGA,TCTTC,GAGTC,GACTC,GTSAC,GAWTC,GCSGC,CASTG,ATGAA,TTCAT,ACGGA,TCCGT,CGCG,CCGC,GCGG,GTAC,AGCT,GCGC,CTAG,GATC,GGCC,CCGG,CATG,RGCY,YATR,ACGT,TGCA,CCDG,CHGG,AATT,CCTC,GAGG,TTAA,ASST,TCGA                                       C                                                                                                                                                                                                                                
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      2   @         #         B   ^   I   =   y   L   8   s   _   @   A  e     +   L         K   Q   _   R  (   X                 C                                                                                                    
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                    	   
   .  	 	      =                            	  d 	      	                    + [                 
     	      > 5   > 3  '   #      ~ L           5 H    	   P  [  )   0 2 .  
  2 @   #   B ^ I = y L 8 s _ @ Ae + L   K Q _ R( X 	  r $ ^ % ;p##Q
0qIfIy H[P$%xb	u		Q

EVkj|"9sy .--/0a36):=k>>@/AdDeDD*E\EGGHHqIJ$LCo28-CH; ?rCFI?n[)ZU*M"%,,]"K36:=Sy
y$'F))T* aKq	G
vHmK,
I,CJ!'22C69=JCJ!'2h!"C07+x	$qCE .-  K%MaKjT	KThq		x		>
G
L5J@v@Gl#!$$&)*,-J..05?b@!APAqC2DDDDE@GHJOKmK1`8!#.%;*+,g1/8H(^3 g!""%%c'+175AACI^3g!"%\'c'+147;z>?5AACIK^3g!"%c'+175AACI(g!c'g!"\'c'Kg!"c'g!c'g!c'	+/1a8EFFG=GHLJuJw1?8$))`	F
!Ms@V.+I.	.C@QN	Xczy	B
{P!!'C@% 'FH6LIczK>)+>p>w>"H	.
.#! ([4#;$) #!o&++FL$>c

(("h#M+v1>8FA1BT.mH
 -CX!g!(*|*?DFFHK$d!"g'a>M+@;)"z1B8GnHnH"h#M+FA1BX.50A?>u>AcI>DvgK2^69'=>4B  ''$+,-F@LJLTLJ'M	jtp2589<AVGGc)TBj}.$cR#0|tIozO		E
N

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Zj~>NCCCfJ+OE->>n		HB!#$t'"(*--Q.,0}5E<?@FFpG@pGkY")+<,q>5L_20k>f #6,y@5F!	,
@UGpGGIcozN
 GI0>Q.,0?CF["(+-
.mIXN "`)GIA]<2AFH> >ELOb)xB+@!L" ,4D'W!':)0C_IJ\w!5; c"Q26k9<> 5oTNk AHz-J|'M	K`fuxjt!"h#
%)M+N+0w1p25?889<>AFAADDVGGI.LA_E2!&'',T.DmH )0n?oAK
"k"-#}#%%%&T)+>TIsSK,xAZDrDxDI/
@bkc#M$CC|el@LkW;		S=;Z !""#B$T$*q+-20m01z1{17B8C8k>q>>@@JAxGG H_HK4L\ c(+>DOE/2b69+=?*r. 9+.4:lE&G
%~Gt	{	C
G

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?+YsSK+C(Bm0>@!	,
!#$t'"(*--Q.,0}5E<?@CFFUGpGGI3$L0L>KGQ*E@L ZfJuAe	=!#B$20m0k>>@JA-%3,z-CLi p"#%C'U(-9.1R8EK   &a
%V_#~ESrGVq	6	J	t	
C
G


y^piSdh{/Iy#d$S,w,J.W01Z8>>>^@@E5FG*GGPHIJ3K7KKKLW;q+m0>@,\36$:=gCFL Z1>mCEE<GfJtJL&Hz!"""&b'i'
),,-0 2>DF<I~B1O58<JLCBq>Z&J4;BKKELOLYL
XA1D4-Q.,0?F8!L%X%c&'MD)i0AA_E	Ip| 2!S#&'',T.DEmHK["G f[,-D=9>,.E_J=$,\36$:=>gCF>J
u !2$=$))*,/\346$:v;=>@AgCDF\GpGHcIlIK@%
:aC+} R!!l"""~##$$%%%&g&x&&& ''''(())Q*]**+=,,/z00<12x3344+666789@:M::Q<<==>>@E@O@AABIDD^EFH.IUIJ6LC~>
 H')24Y69h;"=?@DF&
%~Gt	C
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  A<?xml version="1.0"?><Features nextValidID="34"><Feature recentID="0" name="gRNA scaffold" directionality="1" type="misc_RNA" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="15469-15540" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the &lt;i&gt;Streptococcus pyogenes&lt;/i&gt; CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="At pU3" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="15042-15444" color="#0000ff" type="standard"/></Feature><Feature recentID="2" name="Termination signal" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="15541-15547" color="#0000ff" type="standard"/></Feature><Feature recentID="3" classification="1" name="Feature 18" geneticCode="-1" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="14588-14589" color="#a6acb3" type="standard"/></Feature><Feature recentID="4" name="gRNA scaffold" directionality="1" type="misc_RNA" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="14593-14664" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the &lt;i&gt;Streptococcus pyogenes&lt;/i&gt; CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="At pU6" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="14181-14568" color="#0000ff" type="standard"/></Feature><Feature recentID="6" name="Termination signal" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="14665-14671" color="#0000ff" type="standard"/></Feature><Feature recentID="7" name="gRNA scaffold" directionality="1" type="misc_RNA" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="13729-13804" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the &lt;i&gt;Streptococcus pyogenes&lt;/i&gt; CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="pU3 promoter" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="13306-13704" color="#0000ff" type="standard"/></Feature><Feature recentID="9" name="Termination signal" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="13805-13811" color="#0000ff" type="standard"/></Feature><Feature recentID="10" name="gRNA scaffold" directionality="1" type="misc_RNA" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="12853-12928" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the &lt;i&gt;Streptococcus pyogenes&lt;/i&gt; CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="BsaI" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="12441-12444" color="#0000ff" type="standard"/></Feature><Feature recentID="12" name="AtU6-26 (P)" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="12445-12828" color="#0000ff" type="standard"/></Feature><Feature recentID="13" name="Termination signal" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="12929-12935" color="#0000ff" type="standard"/></Feature><Feature recentID="14" name="FASTRED" directionality="1" translationMW="30402.52" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="18101-18910" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MKLGSKAQDLKDRAQYYGQQHTGGEHDRDRTRGGQHTTMSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDMALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYCDLPSKLGHK*"/></Q></Feature><Feature recentID="15" name="pOLE1::OLE1 " type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="16069-18215" color="#a6acb3" type="standard"/></Feature><Feature recentID="16" name="M13 rev" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6693-6709" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="M13 fwd" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="19170-19186" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="RB T-DNA repeat" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="19382-19406" color="#ffe4c4" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;right border repeat from nopaline C58 T-DNA&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="LB T-DNA repeat" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6394-6418" color="#ffe4c4" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;left border repeat from nopaline C58 T-DNA&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="pVS1 oriV" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3384-3578" color="#ffff00" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;origin of replication for the&amp;nbsp;&lt;i&gt;Pseudomonas&lt;/i&gt; plasmid pVS1 (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/10659714&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/10659714&quot;&gt;Heeb et al., 2000&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="pVS1 RepA" directionality="1" translateFirstCodonAsMet="1" translationMW="39880.55" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2245-3318" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;replication protein from the&amp;nbsp;&lt;i&gt;Pseudomonas&lt;/i&gt; plasmid pVS1 (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/10659714&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/10659714&quot;&gt;Heeb et al., 2000&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILVMRYRNLIEGEASAGS*"/></Q></Feature><Feature recentID="22" name="pVS1 StaA" directionality="1" translationMW="22058.22" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1187-1816" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;stability protein from the&amp;nbsp;&lt;i&gt;Pseudomonas&lt;/i&gt; plasmid pVS1 (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/10659714&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/10659714&quot;&gt;Heeb et al., 2000&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSARDWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIPVQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPILESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI*"/></Q></Feature><Feature recentID="23" name="NOS terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="18909-19160" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;nopaline synthase terminator and poly(A) signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="24" name="SV40 NLS" directionality="1" translationMW="883.15" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" consecutiveNumberingStartsFrom="126" maxRunOn="11849" maxFusedRunOn="11849" detectionMode="exactProteinMatch"><Segment range="11826-11846" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;nuclear localization signal of SV40 (simian virus 40) large T antigen&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="PKKKRKV"/></Q></Feature><Feature recentID="25" name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4248-4836" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="26" name="lac operator" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6669-6685" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;lac&lt;/i&gt; repressor encoded by &lt;i&gt;lacI&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;The &lt;i&gt;lac&lt;/i&gt; repressor binds to the &lt;i&gt;lac&lt;/i&gt; operator to inhibit transcription in &lt;i&gt;E. coli&lt;/i&gt;. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="27" name="CAP binding site" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6595-6616" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;E. coli&lt;/i&gt; catabolite activator protein&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;CAP binding activates transcription in the presence of cAMP.&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="28" name="bom" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3922-4062" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;basis of mobility region from pBR322&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="29" name="lac promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment name="-35" range="6631-6636" color="#ffffff" type="standard"/><Segment range="6637-6654" color="#ffffff" type="standard"/><Segment name="-10" range="6655-6661" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for the &lt;i&gt;E. coli lac&lt;/i&gt; operon&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="30" name="Cas9" directionality="1" translationMW="158441.55" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="11849" maxFusedRunOn="11849" detectionMode="exactProteinMatch"><Segment range="7710-11813" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;generates RNA-guided double strand breaks in DNA&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;Cas9 (Csn1) endonuclease from the&amp;nbsp;&lt;i&gt;Streptococcus pyogenes&lt;/i&gt;&amp;nbsp;Type II CRISPR/Cas system&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"/></Q></Feature><Feature recentID="31" name="SmR" directionality="2" translationMW="29233.19" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="5082-5873" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aadA&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to spectinomycin and streptomycin&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside adenylyltransferase (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/2993813&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/2993813&quot;&gt;Murphy, 1985&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK*"/></Q></Feature><Feature recentID="32" name="attB2" directionality="2" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="15937-15961" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;BP Clonase™&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;mutant version of&amp;nbsp;&lt;i&gt;att&lt;/i&gt;B&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;recombination site for the Gateway® BP reaction&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="33" name="attB1" directionality="1" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="12358-12382" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;BP Clonase™&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;mutant version of&amp;nbsp;&lt;i&gt;att&lt;/i&gt;B&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;recombination site for the Gateway® BP reaction&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  )<?xml version="1.0"?><Primers nextValidID="18"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40" showAdditionalFivePrimeMatches="1" minimumFivePrimeAnnealing="15"/><Primer recentID="0" name="L4440" sequence="AGCGAGTCAGTGAGCGAG" description="L4440 vector, forward primer"><BindingSite location="4076-4093" boundStrand="1" annealedBases="AGCGAGTCAGTGAGCGAG" meltingTemperature="58"><Component hybridizedRange="4076-4093" bases="AGCGAGTCAGTGAGCGAG"/></BindingSite><BindingSite simplified="1" location="4076-4093" boundStrand="1" annealedBases="AGCGAGTCAGTGAGCGAG" meltingTemperature="58"><Component hybridizedRange="4076-4093" bases="AGCGAGTCAGTGAGCGAG"/></BindingSite></Primer><Primer recentID="1" name="M13 Reverse" sequence="CAGGAAACAGCTATGAC" description="In lacZ gene. Also called M13-rev"><BindingSite location="6692-6708" boundStrand="0" annealedBases="CAGGAAACAGCTATGAC" meltingTemperature="49"><Component hybridizedRange="6692-6708" bases="CAGGAAACAGCTATGAC"/></BindingSite><BindingSite simplified="1" location="6692-6708" boundStrand="0" annealedBases="CAGGAAACAGCTATGAC" meltingTemperature="49"><Component hybridizedRange="6692-6708" bases="CAGGAAACAGCTATGAC"/></BindingSite></Primer><Primer recentID="2" name="M13/pUC Reverse" sequence="AGCGGATAACAATTTCACACAGG" description="In lacZ gene"><BindingSite location="6673-6695" boundStrand="0" annealedBases="AGCGGATAACAATTTCACACAGG" meltingTemperature="58"><Component hybridizedRange="6673-6695" bases="AGCGGATAACAATTTCACACAGG"/></BindingSite><BindingSite simplified="1" location="6673-6695" boundStrand="0" annealedBases="AGCGGATAACAATTTCACACAGG" meltingTemperature="58"><Component hybridizedRange="6673-6695" bases="AGCGGATAACAATTTCACACAGG"/></BindingSite></Primer><Primer recentID="3" name="pBR322ori-F" sequence="GGGAAACGCCTGGTATCTTT" description="pBR322 origin, forward primer"><BindingSite location="4327-4346" boundStrand="1" annealedBases="GGGAAACGCCTGGTATCTTT" meltingTemperature="56"><Component hybridizedRange="4327-4346" bases="GGGAAACGCCTGGTATCTTT"/></BindingSite><BindingSite simplified="1" location="4327-4346" boundStrand="1" annealedBases="GGGAAACGCCTGGTATCTTT" meltingTemperature="56"><Component hybridizedRange="4327-4346" bases="GGGAAACGCCTGGTATCTTT"/></BindingSite></Primer><Primer recentID="4" name="pGEX 3'" sequence="CCGGGAGCTGCATGTGTCAGAGG" description="pGEX vectors, reverse primer"><BindingSite location="3813-3835" boundStrand="1" annealedBases="CCGGGAGCTGCATGTGTCAGAGG" meltingTemperature="66"><Component hybridizedRange="3813-3835" bases="CCGGGAGCTGCATGTGTCAGAGG"/></BindingSite><BindingSite simplified="1" location="3813-3835" boundStrand="1" annealedBases="CCGGGAGCTGCATGTGTCAGAGG" meltingTemperature="66"><Component hybridizedRange="3813-3835" bases="CCGGGAGCTGCATGTGTCAGAGG"/></BindingSite></Primer><Primer recentID="5" name="pRS-marker" sequence="CGGCATCAGAGCAGATTGTA" description="pRS vectors, use to sequence yeast selectable marker"><BindingSite location="3976-3995" boundStrand="0" annealedBases="CGGCATCAGAGCAGATTGTA" meltingTemperature="54"><Component hybridizedRange="3976-3995" bases="CGGCATCAGAGCAGATTGTA"/></BindingSite><BindingSite simplified="1" location="3976-3995" boundStrand="0" annealedBases="CGGCATCAGAGCAGATTGTA" meltingTemperature="54"><Component hybridizedRange="3976-3995" bases="CGGCATCAGAGCAGATTGTA"/></BindingSite></Primer><Primer recentID="6" name="oRU906" sequence="GAGTCTATGATCAAGTAATTATGC" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2023-01-01T01:14:45Z"><BindingSite location="12282-12305" boundStrand="0" annealedBases="GAGTCTATGATCAAGTAATTATGC" meltingTemperature="52"><Component hybridizedRange="12282-12305" bases="GAGTCTATGATCAAGTAATTATGC"/></BindingSite><BindingSite simplified="1" location="12282-12305" boundStrand="0" annealedBases="GAGTCTATGATCAAGTAATTATGC" meltingTemperature="52"><Component hybridizedRange="12282-12305" bases="GAGTCTATGATCAAGTAATTATGC"/></BindingSite></Primer><Primer recentID="7" name="&gt;Cas9_NUB_gRNA7_fw " sequence="ATTGGGATCTTGCCTTGACGGTGG" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2022-12-20T14:31:45Z"><BindingSite location="12828-12851" boundStrand="0" annealedBases="ATTGGGATCTTGCCTTGACGGTGG" meltingTemperature="63"><Component hybridizedRange="12828-12851" bases="ATTGGGATCTTGCCTTGACGGTGG"/></BindingSite><BindingSite simplified="1" location="12828-12851" boundStrand="0" annealedBases="ATTGGGATCTTGCCTTGACGGTGG" meltingTemperature="63"><Component hybridizedRange="12828-12851" bases="ATTGGGATCTTGCCTTGACGGTGG"/></BindingSite></Primer><Primer recentID="8" name="&gt;Cas9_NUB_gRNA7_rv " sequence="aaacCCACCGTCAAGGCAAGATCC" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2022-12-20T14:32:12Z"><BindingSite location="12832-12855" boundStrand="1" annealedBases="aaacCCACCGTCAAGGCAAGATCC" meltingTemperature="64"><Component hybridizedRange="12832-12855" bases="aaacCCACCGTCAAGGCAAGATCC"/></BindingSite><BindingSite simplified="1" location="12832-12855" boundStrand="1" annealedBases="aaacCCACCGTCAAGGCAAGATCC" meltingTemperature="64"><Component hybridizedRange="12832-12855" bases="aaacCCACCGTCAAGGCAAGATCC"/></BindingSite></Primer><Primer recentID="9" name="&gt;Cas9_NUB_gRNA5_fw " sequence="GTCAAGCATGGAACTCAAAGTGGA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2022-12-20T14:21:59Z"><BindingSite location="13704-13727" boundStrand="0" annealedBases="GTCAAGCATGGAACTCAAAGTGGA" meltingTemperature="58"><Component hybridizedRange="13704-13727" bases="GTCAAGCATGGAACTCAAAGTGGA"/></BindingSite><BindingSite simplified="1" location="13704-13727" boundStrand="0" annealedBases="GTCAAGCATGGAACTCAAAGTGGA" meltingTemperature="58"><Component hybridizedRange="13704-13727" bases="GTCAAGCATGGAACTCAAAGTGGA"/></BindingSite></Primer><Primer recentID="10" name="&gt;Cas9_NUB_gRNA5_rv " sequence="aaacTCCACTTTGAGTTCCATGCT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2022-12-20T14:22:44Z"><BindingSite location="13708-13731" boundStrand="1" annealedBases="aaacTCCACTTTGAGTTCCATGCT" meltingTemperature="59"><Component hybridizedRange="13708-13731" bases="aaacTCCACTTTGAGTTCCATGCT"/></BindingSite><BindingSite simplified="1" location="13708-13731" boundStrand="1" annealedBases="aaacTCCACTTTGAGTTCCATGCT" meltingTemperature="59"><Component hybridizedRange="13708-13731" bases="aaacTCCACTTTGAGTTCCATGCT"/></BindingSite></Primer><Primer recentID="11" name="&gt;Cas9_NUB_gRNA6_rv " sequence="AaaCTCCATCTTCACTCTCTCAAC" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2022-12-20T14:42:52Z"><BindingSite location="14568-14591" boundStrand="1" annealedBases="AaaCTCCATCTTCACTCTCTCAAC" meltingTemperature="56"><Component hybridizedRange="14568-14591" bases="AaaCTCCATCTTCACTCTCTCAAC"/></BindingSite><BindingSite simplified="1" location="14568-14591" boundStrand="1" annealedBases="AaaCTCCATCTTCACTCTCTCAAC" meltingTemperature="56"><Component hybridizedRange="14568-14591" bases="AaaCTCCATCTTCACTCTCTCAAC"/></BindingSite></Primer><Primer recentID="12" name="&gt;Cas9_NUB_gRNA6_fw " sequence="ATTGGTTGAGAGAGTGAAGATGGA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2022-12-20T14:43:08Z"><BindingSite location="14564-14587" boundStrand="0" annealedBases="ATTGGTTGAGAGAGTGAAGATGGA" meltingTemperature="56"><Component hybridizedRange="14564-14587" bases="ATTGGTTGAGAGAGTGAAGATGGA"/></BindingSite><BindingSite simplified="1" location="14564-14587" boundStrand="0" annealedBases="ATTGGTTGAGAGAGTGAAGATGGA" meltingTemperature="56"><Component hybridizedRange="14564-14587" bases="ATTGGTTGAGAGAGTGAAGATGGA"/></BindingSite></Primer><Primer recentID="13" name="&gt;Cas9_NUB_gRNA8_rv " sequence="AaaCCCGCTGTCTTCACCGTTTCC" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2022-12-20T14:48:23Z"><BindingSite location="15444-15467" boundStrand="1" annealedBases="AaaCCCGCTGTCTTCACCGTTTCC" meltingTemperature="64"><Component hybridizedRange="15444-15467" bases="AaaCCCGCTGTCTTCACCGTTTCC"/></BindingSite><BindingSite simplified="1" location="15444-15467" boundStrand="1" annealedBases="AaaCCCGCTGTCTTCACCGTTTCC" meltingTemperature="64"><Component hybridizedRange="15444-15467" bases="AaaCCCGCTGTCTTCACCGTTTCC"/></BindingSite></Primer><Primer recentID="14" name="&gt;Cas9_NUB_gRNA8_fw " sequence="GTCAGGAAACGGTGAAGACAGCGG" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2022-12-20T14:48:39Z"><BindingSite location="15440-15463" boundStrand="0" annealedBases="GTCAGGAAACGGTGAAGACAGCGG" meltingTemperature="63"><Component hybridizedRange="15440-15463" bases="GTCAGGAAACGGTGAAGACAGCGG"/></BindingSite><BindingSite simplified="1" location="15440-15463" boundStrand="0" annealedBases="GTCAGGAAACGGTGAAGACAGCGG" meltingTemperature="63"><Component hybridizedRange="15440-15463" bases="GTCAGGAAACGGTGAAGACAGCGG"/></BindingSite></Primer><Primer recentID="15" name="M13 Forward" sequence="TGTAAAACGACGGCCAGT" description="In lacZ gene. Also called M13-F20 or M13 (-21) Forward"><BindingSite location="19169-19186" boundStrand="1" annealedBases="TGTAAAACGACGGCCAGT" meltingTemperature="57"><Component hybridizedRange="19169-19186" bases="TGTAAAACGACGGCCAGT"/></BindingSite><BindingSite simplified="1" location="19169-19186" boundStrand="1" annealedBases="TGTAAAACGACGGCCAGT" meltingTemperature="57"><Component hybridizedRange="19169-19186" bases="TGTAAAACGACGGCCAGT"/></BindingSite></Primer><Primer recentID="16" name="M13/pUC Forward" sequence="CCCAGTCACGACGTTGTAAAACG" description="In lacZ gene"><BindingSite location="19178-19200" boundStrand="1" annealedBases="CCCAGTCACGACGTTGTAAAACG" meltingTemperature="61"><Component hybridizedRange="19178-19200" bases="CCCAGTCACGACGTTGTAAAACG"/></BindingSite><BindingSite simplified="1" location="19178-19200" boundStrand="1" annealedBases="CCCAGTCACGACGTTGTAAAACG" meltingTemperature="61"><Component hybridizedRange="19178-19200" bases="CCCAGTCACGACGTTGTAAAACG"/></BindingSite></Primer><Primer recentID="17" name="oRU908" sequence="GCTTGCATGCCTGCAGGTCGACTCT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2023-01-01T01:14:34Z"><BindingSite location="16046-16070" boundStrand="1" annealedBases="GCTTGCATGCCTGCAGGTCGACTCT" meltingTemperature="66"><Component hybridizedRange="16046-16070" bases="GCTTGCATGCCTGCAGGTCGACTCT"/></BindingSite><BindingSite simplified="1" location="16046-16070" boundStrand="1" annealedBases="GCTTGCATGCCTGCAGGTCGACTCT" meltingTemperature="66"><Component hybridizedRange="16046-16070" bases="GCTTGCATGCCTGCAGGTCGACTCT"/></BindingSite></Primer></Primers>
  A<Notes>
<UUID>3e827717-963a-4495-a836-b4dd0f071d9a</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="9:41:29">2023.2.14</Created>
<LastModified UTC="9:41:29">2023.2.14</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  Y              AG Becker enzyme                                                                                            d     CHO  d               E              Y? P                                                                                                                                                              <?xml version="1.0"?><CustomEnzymeSets><CustomEnzymeSet type="0" name="AG Becker enzyme" enzymeNames="AhdI BamHI BbsI BsaI BsmBI ClaI EcoRI HindIII KpnI MluI NcoI NotI NspI PmeI PstI SacI SalI SmaI SpeI TaqI XbaI XhoI"/></CustomEnzymeSets>
   3<?xml version="1.0"?><EnzymeVisibilities vals=""/>
