Addgene pYAMTr2GCsgScLEU2 Sequencing Result - Sequence Analyzer Skip to main content
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Sequence Analyzer: pYAMTr2GCsgScLEU2 Sequencing Result


Map View

PshAI (12,763) Neo-R (10,865 .. 10,884) NeoR/KanR RsrII (10,290) Neo-F (10,255 .. 10,274) AgeI (10,049) RB T-DNA repeat lac operator M13 rev ApaI (9679) PspOMI (9675) pBluescriptKS (9654 .. 9670) AbsI - PspXI (9666) KS primer GPDpro-F (9049 .. 9072) BsaI (8740) SgrDI (8536) BlpI (8224) BsrGI (7344) PmlI (14) pBRforEco (1416 .. 1434) pGEX 3' (1472 .. 1494) pRS-marker (1594 .. 1613) ApaLI (1620) NruI (2095) Kan-R (2080 .. 2099) AsiSI (2438) F1ori-F (3244 .. 3265) F1ori-R (3456 .. 3475) M13/pUC Forward (3688 .. 3710) M13 Forward (3702 .. 3719) M13 fwd T7 (3729 .. 3748) NotI (3773) pBluescriptSK (3780 .. 3796) SK primer TspMI - XmaI (3798) SmaI (3800) CAP binding site lac operator M13 rev T3 promoter NheI (4187) BmtI (4191) gRNA scaffold SUP4 terminator ADH1 terminator pHybLex-R (4667 .. 4691) SV40 NLS pYAMTr2GCsgScLEU2 12,843 bp

Sequence View

PmlI 5ʹ 3ʹ 60 2μ ori g t c c t t t t c a t c a c g t g a g t g c c a c c t g a a c g a a g c a t c t g t g c t t c a t t t t g t a g a a c a c a g g a a a a g t a g t g c a c t c a c g g t g g a c t t g c t t c g t a g a c a c g a a g t a a a a c a t c t t g t

120 2μ ori a a a a t g c a a c g c g a g a g c g c t a a t t t t t c a a a c a a a g a a t c t g a g c t g c a t t t t t a c a g a t t t t a c g t t g c g c t c t c g c g a t t a a a a a g t t t g t t t c t t a g a c t c g a c g t a a a a a t g t c t

180 2μ ori a c a g a a a t g c a a c g c g a a a g c g c t a t t t t a c c a a c g a a g a a t c t g t g c t t c a t t t t t g t a t g t c t t t a c g t t g c g c t t t c g c g a t a a a a t g g t t g c t t c t t a g a c a c g a a g t a a a a a c a t

240 2μ ori a a a c a a a a a t g c a a c g c g a g a g c g c t a a t t t t t c a a a c a a a g a a t c t g a g c t g c a t t t t t t t t g t t t t t a c g t t g c g c t c t c g c g a t t a a a a a g t t t g t t t c t t a g a c t c g a c g t a a a a a

300 2μ ori a c a g a a c a g a a a t g c a a c g c g a g a g c g c t a t t t t a c c a a c a a a g a a t c t a t a c t t c t t t t t g t c t t g t c t t t a c g t t g c g c t c t c g c g a t a a a a t g g t t g t t t c t t a g a t a t g a a g a a a a

360 2μ ori t t g t t c t a c a a a a a t g c a t c c c g a g a g c g c t a t t t t t c t a a c a a a g c a t c t t a g a t t a c t a a c a a g a t g t t t t t a c g t a g g g c t c t c g c g a t a a a a a g a t t g t t t c g t a g a a t c t a a t g a

420 2μ ori t t t t t t c t c c t t t g t g c g c t c t a t a a t g c a g t c t c t t g a t a a c t t t t t g c a c t g t a g g t c a a a a a a g a g g a a a c a c g c g a g a t a t t a c g t c a g a g a a c t a t t g a a a a a c g t g a c a t c c a g

480 2μ ori c g t t a a g g t t a g a a g a a g g c t a c t t t g g t g t c t a t t t t c t c t t c c a t a a a a a a a g c c t g a g c a a t t c c a a t c t t c t t c c g a t g a a a c c a c a g a t a a a a g a g a a g g t a t t t t t t t c g g a c t

540 2μ ori c t c c a c t t c c c g c g t t t a c t g a t t a c t a g c g a a g c t g c g g g t g c a t t t t t t c a a g a t a a a g a g g t g a a g g g c g c a a a t g a c t a a t g a t c g c t t c g a c g c c c a c g t a a a a a a g t t c t a t t t

600 2μ ori g g c a t c c c c g a t t a t a t t c t a t a c c g a t g t g g a t t g c g c a t a c t t t g t g a a c a g a a a g t g c c g t a g g g g c t a a t a t a a g a t a t g g c t a c a c c t a a c g c g t a t g a a a c a c t t g t c t t t c a c

660 2μ ori a t a g c g t t g a t g a t t c t t c a t t g g t c a g a a a a t t a t g a a c g g t t t c t t c t a t t t t g t c t c t a t c g c a a c t a c t a a g a a g t a a c c a g t c t t t t a a t a c t t g c c a a a g a a g a t a a a a c a g a g

720 2μ ori t a t a t a c t a c g t a t a g g a a a t g t t t a c a t t t t c g t a t t g t t t t c g a t t c a c t c t a t g a a t a t a t a t g a t g c a t a t c c t t t a c a a a t g t a a a a g c a t a a c a a a a g c t a a g t g a g a t a c t t a

780 2μ ori a g t t c t t a c t a c a a t t t t t t t g t c t a a a g a g t a a t a c t a g a g a t a a a c a t a a a a a a t g t a t c a a g a a t g a t g t t a a a a a a a c a g a t t t c t c a t t a t g a t c t c t a t t t g t a t t t t t t a c a t

840 2μ ori g a g g t c g a g t t t a g a t g c a a g t t c a a g g a g c g a a a g g t g g a t g g g t a g g t t a t a t a g g g a c t c c a g c t c a a a t c t a c g t t c a a g t t c c t c g c t t t c c a c c t a c c c a t c c a a t a t a t c c c t

900 2μ ori t a t a g c a c a g a g a t a t a t a g c a a a g a g a t a c t t t t g a g c a a t g t t t g t g g a a g c g g t a t t a t a t c g t g t c t c t a t a t a t c g t t t c t c t a t g a a a a c t c g t t a c a a a c a c c t t c g c c a t a a

960 2μ ori c g c a a t a t t t t a g t a g c t c g t t a c a g t c c g g t g c g t t t t t g g t t t t t t g a a a g t g c g t c t g c g t t a t a a a a t c a t c g a g c a a t g t c a g g c c a c g c a a a a a c c a a a a a a c t t t c a c g c a g a

1020 2μ ori t c a g a g c g c t t t t g g t t t t c a a a a g c g c t c t g a a g t t c c t a t a c t t t c t a g c t a g a g a a t a g t c t c g c g a a a a c c a a a a g t t t t c g c g a g a c t t c a a g g a t a t g a a a g a t c g a t c t c t t a

1080 2μ ori a g g a a c t t c g g a a t a g g a a c t t c a a a g c g t t t c c g a a a a c g a g c g c t t c c g a a a a t g c a a t c c t t g a a g c c t t a t c c t t g a a g t t t c g c a a a g g c t t t t g c t c g c g a a g g c t t t t a c g t t

1140 2μ ori c g c g a g c t g c g c a c a t a c a g c t c a c t g t t c a c g t c g c a c c t a t a t c t g c g t g t t g c c t g t g c g c t c g a c g c g t g t a t g t c g a g t g a c a a g t g c a g c g t g g a t a t a g a c g c a c a a c g g a c a

1200 2μ ori a t a t a t a t a t a c a t g a g a a g a a c g g c a t a g t g c g t g t t t a t g c t t a a a t g c g t a c t t a t a t a t a t a t a t a t g t a c t c t t c t t g c c g t a t c a c g c a c a a a t a c g a a t t t a c g c a t g a a t a t

1260 2μ ori t g c g t c t a t t t a t g t a g g a t g a a a g g t a g t c t a g t a c c t c c t g t g a t a t t a t c c c a t t c c a c g c a g a t a a a t a c a t c c t a c t t t c c a t c a g a t c a t g g a g g a c a c t a t a a t a g g g t a a g g

1320 2μ ori a t g c g g g g t a t c g t a t g c t t c c t t c a g c a c t a c c c t t t a g c t g t t c t a t a t g c t g c c a c t t a c g c c c c a t a g c a t a c g a a g g a a g t c g t g a t g g g a a a t c g a c a a g a t a t a c g a c g g t g a

1380 2μ ori c c t c a a t t g g a t t a g t c t c a t c c t t c a a t g c t a t c a t t t c c t t t g a t a t t g g a t c a t a c t g g a g t t a a c c t a a t c a g a g t a g g a a g t t a c g a t a g t a a a g g a a a c t a t a a c c t a g t a t g a

1440 A A T A G G C G T A T C A C G A G G C pBRforEco a a g a a a c c a t t a t t a t c a t g a c a t t a a c c t a t a a a a a t a g g c g t a t c a c g a g g c c c t t t c t t c t t t g g t a a t a a t a g t a c t g t a a t t g g a t a t t t t t a t c c g c a t a g t g c t c c g g g a a a g

1500 G G A G A C T G T G T A C G T C G A G G G C C pGEX 3' g t c t c g c g c g t t t c g g t g a t g a c g g t g a a a a c c t c t g a c a c a t g c a g c t c c c g g a g a c g g c a g a g c g c g c a a a g c c a c t a c t g c c a c t t t t g g a g a c t g t g t a c g t c g a g g g c c t c t g c c

1560 t c a c a g c t t g t c t g t a a g c g g a t g c c g g g a g c a g a c a a g c c c g t c a g g g c g c g t c a g c g g a g t g t c g a a c a g a c a t t c g c c t a c g g c c c t c g t c t g t t c g g g c a g t c c c g c g c a g t c g c c

1620 C G G C A T C A G A G C A G A T T G T A pRS-marker g t g t t g g c g g g t g t c g g g g c t g g c t t a a c t a t g c g g c a t c a g a g c a g a t t g t a c t g a g a g c a c a a c c g c c c a c a g c c c c g a c c g a a t t g a t a c g c c g t a g t c t c g t c t a a c a t g a c t c t c

ApaLI 1680 kanMX TEF promoter t g c a c c a t a g t t t a g c t t g c c t c g t c c c c g c c g g g t c a c c c g g c c a g c g a c a t g g a g g c c a c g t g g t a t c a a a t c g a a c g g a g c a g g g g c g g c c c a g t g g g c c g g t c g c t g t a c c t c c g g

1740 kanMX TEF promoter c a g a a t a c c c t c c t t g a c a g t c t t g a c g t g c g c a g c t c a g g g g c a t g a t g t g a c t g t c g c g t c t t a t g g g a g g a a c t g t c a g a a c t g c a c g c g t c g a g t c c c c g t a c t a c a c t g a c a g c g

1800 kanMX TEF promoter c c g t a c a t t t a g c c c a t a c a t c c c c a t g t a t a a t c a t t t g c a t c c a t a c a t t t t g a t g g c g g c a t g t a a a t c g g g t a t g t a g g g g t a c a t a t t a g t a a a c g t a g g t a t g t a a a a c t a c c g

1860 kanMX TEF promoter c g c a c g g c g c g a a g c a a a a a t t a c g g c t c c t c g c t g c a g a c c t g c g a g c a g g g a a a c g c t g c g t g c c g c g c t t c g t t t t t a a t g c c g a g g a g c g a c g t c t g g a c g c t c g t c c c t t t g c g a

1920 kanMX TEF promoter c c c c t c a c a g a c g c g t t g a a t t g t c c c c a c g c c g c g c c c c t g t a g a g a a a t a t a a a a g g t g g g g a g t g t c t g c g c a a c t t a a c a g g g g t g c g g c g c g g g g a c a t c t c t t t a t a t t t t c c a

1980 kanMX TEF promoter t a g g a t t t g c c a c t g a g g t t c t t c t t t c a t a t a c t t c c t t t t a a a a t c t t g c t a g g a t a c a t c c t a a a c g g t g a c t c c a a g a a g a a a g t a t a t g a a g g a a a a t t t t a g a a c g a t c c t a t g

2040 kanMX 1 5 MGKEKTHVS TEF promoter KanR MGKEKTHVS a g t t c t c a c a t c a c a t c c g a a c a t a a a c a a c c a t g g g t a a g g a a a a g a c t c a c g t t t c g a t c a a g a g t g t a g t g t a g g c t t g t a t t t g t t g g t a c c c a t t c c t t t t c t g a g t g c a a a g c t

NruI 2100 kanMX 10 15 20 25 RPRLNSNMDADLYGYKWARD KanR C C A T A T T T A C C C G A G C G C T A Kan-R RPRLNSNMDADLYGYKWARD g g c c g c g a t t a a a t t c c a a c a t g g a t g c t g a t t t a t a t g g g t a t a a a t g g g c t c g c g a t a c c g g c g c t a a t t t a a g g t t g t a c c t a c g a c t a a a t a t a c c c a t a t t t a c c c g a g c g c t a t

2160 kanMX 30 35 40 45 NVGQSGATIYRLYGKPDAPE KanR NVGQSGATIYRLYGKPDAPE a t g t c g g g c a a t c a g g t g c g a c a a t c t a t c g a t t g t a t g g g a a g c c c g a t g c g c c a g a g t t a c a g c c c g t t a g t c c a c g c t g t t a g a t a g c t a a c a t a c c c t t c g g g c t a c g c g g t c t c a

2220 kanMX 50 55 60 65 LFLKHGKGSVANDVTDEMVR KanR LFLKHGKGSVANDVTDEMVR t g t t t c t g a a a c a t g g c a a a g g t a g c g t t g c c a a t g a t g t t a c a g a t g a g a t g g t c a g a c a c a a a g a c t t t g t a c c g t t t c c a t c g c a a c g g t t a c t a c a a t g t c t a c t c t a c c a g t c t g

2280 kanMX 70 75 80 85 LNWLTEFMPLPTIKHFIRTP KanR LNWLTEFMPLPTIKHFIRTP t a a a c t g g c t g a c g g a a t t t a t g c c t c t t c c g a c c a t c a a g c a t t t t a t c c g t a c t c c t g a t t t g a c c g a c t g c c t t a a a t a c g g a g a a g g c t g g t a g t t c g t a a a a t a g g c a t g a g g a c

2340 kanMX 90 95 100 105 DDAWLLTTAIPGKTAFQVLE KanR DDAWLLTTAIPGKTAFQVLE a t g a t g c a t g g t t a c t c a c c a c t g c g a t c c c c g g c a a a a c a g c a t t c c a g g t a t t a g a a g t a c t a c g t a c c a a t g a g t g g t g a c g c t a g g g g c c g t t t t g t c g t a a g g t c c a t a a t c t t c

2400 kanMX 110 115 120 125 EYPDSGENIVDALAVFLRRL KanR EYPDSGENIVDALAVFLRRL a a t a t c c t g a t t c a g g t g a a a a t a t t g t t g a t g c g c t g g c a g t g t t c c t g c g c c g g t t g c t t a t a g g a c t a a g t c c a c t t t t a t a a c a a c t a c g c g a c c g t c a c a a g g a c g c g g c c a a c g

AsiSI 2460 kanMX 130 135 140 145 HSIPVCNCPFNSDRVFRLAQ KanR HSIPVCNCPFNSDRVFRLAQ a t t c g a t t c c t g t t t g t a a t t g t c c t t t t a a c a g c g a t c g c g t a t t t c g t c t c g c t c a g g t a a g c t a a g g a c a a a c a t t a a c a g g a a a a t t g t c g c t a g c g c a t a a a g c a g a g c g a g t c c

2520 kanMX 150 155 160 165 AQSRMNNGLVDASDFDDERN KanR AQSRMNNGLVDASDFDDERN c g c a a t c a c g a a t g a a t a a c g g t t t g g t t g a t g c g a g t g a t t t t g a t g a c g a g c g t a a t g g c g t t a g t g c t t a c t t a t t g c c a a a c c a a c t a c g c t c a c t a a a a c t a c t g c t c g c a t t a c

2580 kanMX 170 175 180 185 GWPVEQVWKEMHKLLPFSPD KanR GWPVEQVWKEMHKLLPFSPD g c t g g c c t g t t g a a c a a g t c t g g a a a g a a a t g c a t a a g c t t t t g c c a t t c t c a c c g g a t t c g a c c g g a c a a c t t g t t c a g a c c t t t c t t t a c g t a t t c g a a a a c g g t a a g a g t g g c c t a a

2640 kanMX 190 195 200 205 SVVTHGDFSLDNLIFDEGKL KanR SVVTHGDFSLDNLIFDEGKL c a g t c g t c a c t c a t g g t g a t t t c t c a c t t g a t a a c c t t a t t t t t g a c g a g g g g a a a t t a a g t c a g c a g t g a g t a c c a c t a a a g a g t g a a c t a t t g g a a t a a a a a c t g c t c c c c t t t a a t t

2700 kanMX 210 215 220 225 IGCIDVGRVGIADRYQDLAI KanR IGCIDVGRVGIADRYQDLAI t a g g t t g t a t t g a t g t t g g a c g a g t c g g a a t c g c a g a c c g a t a c c a g g a t c t t g c c a t c c a t c c a a c a t a a c t a c a a c c t g c t c a g c c t t a g c g t c t g g c t a t g g t c c t a g a a c g g t a g g

2760 kanMX 230 235 240 245 LWNCLGEFSPSLQKRLFQKY KanR LWNCLGEFSPSLQKRLFQKY t a t g g a a c t g c c t c g g t g a g t t t t c t c c t t c a t t a c a g a a a c g g c t t t t t c a a a a a t a t g a t a c c t t g a c g g a g c c a c t c a a a a g a g g a a g t a a t g t c t t t g c c g a a a a a g t t t t t a t a c

2820 kanMX 250 255 260 265 GIDNPDMNKLQFHLMLDEFF KanR GIDNPDMNKLQFHLMLDEFF g t a t t g a t a a t c c t g a t a t g a a t a a a t t g c a g t t t c a t t t g a t g c t c g a t g a g t t t t t c t c a t a a c t a t t a g g a c t a t a c t t a t t t a a c g t c a a a g t a a a c t a c g a g c t a c t c a a a a a g a

2880 kanMX 270 * KanR TEF terminator * a a t c a g t a c t g a c a a t a a a a a g a t t c t t g t t t t c a a g a a c t t g t c a t t t g t a t a g t t t t t t t a g t c a t g a c t g t t a t t t t t c t a a g a a c a a a a g t t c t t g a a c a g t a a a c a t a t c a a a a a

2940 kanMX TEF terminator t t a t a t t g t a g t t g t t c t a t t t t a a t c a a a t g t t a g c g t g a t t t a t a t t t t t t t t c g c c t a a t a t a a c a t c a a c a a g a t a a a a t t a g t t t a c a a t c g c a c t a a a t a t a a a a a a a a g c g g a

3000 kanMX TEF terminator c g a c a t c a t c t g c c c a g a t g c g a a g t t a a g t g c g c a g a a a g t a a t a t c a t g c g t c a a t c g g c t g t a g t a g a c g g g t c t a c g c t t c a a t t c a c g c g t c t t t c a t t a t a g t a c g c a g t t a g c

3060 kanMX TEF terminator t a t g t g a a t g c t g g t c g c t a t a c t g c t g t c g a t t c g a t a c t a a c g c c g c c a t c c a g t g t c a t a c a c t t a c g a c c a g c g a t a t g a c g a c a g c t a a g c t a t g a t t g c g g c g g t a g g t c a c a g

3120 f1 ori g a a a a c a a t a c c g c a c a g a t g c g t a a g g a g a a a a t a c c g c a t c a g g a a a t t g t a a g c g t t c t t t t g t t a t g g c g t g t c t a c g c a t t c c t c t t t t a t g g c g t a g t c c t t t a a c a t t c g c a a

3180 f1 ori a a t a t t t t g t t a a a a t t c g c g t t a a a t t t t t g t t a a a t c a g c t c a t t t t t t a a c c a a t a g t t a t a a a a c a a t t t t a a g c g c a a t t t a a a a a c a a t t t a g t c g a g t a a a a a a t t g g t t a t c

3240 f1 ori g c c g a a a t c g g c a a a a t c c c t t a t a a a t c a a a a g a a t a g a c c g a g a t a g g g t t g a g t g t t c g g c t t t a g c c g t t t t a g g g a a t a t t t a g t t t t c t t a t c t g g c t c t a t c c c a a c t c a c a a

3300 f1 ori G G T C A A A C C T T G T T C T C A G G T G F1ori-F g t t c c a g t t t g g a a c a a g a g t c c a c t a t t a a a g a a c g t g g a c t c c a a c g t c a a a g g g c g a c a a g g t c a a a c c t t g t t c t c a g g t g a t a a t t t c t t g c a c c t g a g g t t g c a g t t t c c c g c t

3360 f1 ori a a a a c c g t c t a t c a g g g c g a t g g c c c a c t a c g t g a a c c a t c a c c c t a a t c a a g t t t t t t g t t t t g g c a g a t a g t c c c g c t a c c g g g t g a t g c a c t t g g t a g t g g g a t t a g t t c a a a a a a c

3420 f1 ori g g g t c g a g g t g c c g t a a a g c a c t a a a t c g g a a c c c t a a a g g g a g c c c c c g a t t t a g a g c t c c c a g c t c c a c g g c a t t t c g t g a t t t a g c c t t g g g a t t t c c c t c g g g g g c t a a a t c t c g a

3480 f1 ori A G G G A A G A A A G C G A A A G G A G F1ori-R t g a c g g g g a a a g c c g g c g a a c g t g g c g a g a a a g g a a g g g a a g a a a g c g a a a g g a g c g g g c a c t g c c c c t t t c g g c c g c t t g c a c c g c t c t t t c c t t c c c t t c t t t c g c t t t c c t c g c c c g

3540 f1 ori g c t a g g g c g c t g g c a a g t g t a g c g g t c a c g c t g c g c g t a a c c a c c a c a c c c g c c g c g c t t c g a t c c c g c g a c c g t t c a c a t c g c c a g t g c g a c g c g c a t t g g t g g t g t g g g c g g c g c g a a

3600 f1 ori a a t g c g c c g c t a c a g g g c g c g t c c a t t c g c c a t t c a g g c t g c g c a a c t g t t g g g a a g g g c t t a c g c g g c g a t g t c c c g c g c a g g t a a g c g g t a a g t c c g a c g c g t t g a c a a c c c t t c c c g

3660 g a t c g g t g c g g g c c t c t t c g c t a t t a c g c c a g c t g g c g a a a g g g g g a t g t g c t g c a a g g c c t a g c c a c g c c c g g a g a a g c g a t a a t g c g g t c g a c c g c t t t c c c c c t a c a c g a c g t t c c g

3720 M13 fwd C C C A G T C A C G A C G T T G T A A A A C G M13/pUC Forward T G T A A A A C G A C G G C C A G T M13 Forward g a t t a a g t t g g g t a a c g c c a g g g t t t t c c c a g t c a c g a c g t t g t a a a a c g a c g g c c a g t g c t a a t t c a a c c c a t t g c g g t c c c a a a a g g g t c a g t g c t g c a a c a t t t t g c t g c c g g t c a c

NotI 3780 T7 promoter SK primer T A A T A C G A C T C A C T A T A G G G T7 T pBluescriptSK a g c g c g c g t a a t a c g a c t c a c t a t a g g g c g a a t t g g a g c t c c a c c g c g g t g g c g g c c g c t t c g c g c g c a t t a t g c t g a g t g a t a t c c c g c t t a a c c t c g a g g t g g c g c c a c c g c c g g c g a

TspMIXmaISmaI 3840 SK primer C T A G A A C T A G T G G A T C pBluescriptSK c t a g a a c t a g t g g a t c c c c c g g g c t g c a g g a a t t c g a t c t g g c a c g a c a g g t t t c c c g a c g a t c t t g a t c a c c t a g g g g g c c c g a c g t c c t t a a g c t a g a c c g t g c t g t c c a a a g g g c t g

3900 CAP binding site t g g a a a g c g g g c a g t g a g c g c a a c g c a a t t a a t g t g a g t t a g c t c a c t c a t t a g g c a c c c a c c t t t c g c c c g t c a c t c g c g t t g c g t t a a t t a c a c t c a a t c g a g t g a g t a a t c c g t g g g

3960 -35 -10 lac promoter lac operator c a g g c t t t a c a c t t t a t g c t t c c g g c t c g t a t g t t g t g t g g a a t t g t g a g c g g a t a a c a a g t c c g a a a t g t g a a a t a c g a a g g c c g a g c a t a c a a c a c a c c t t a a c a c t c g c c t a t t g t t

4020 M13 rev T3 promoter t t t c a c a c a g g a a a c a g c t a t g a c c a t g a t t a c g c c a a g c g c g c a a t t a a c c c t c a c t a a a a a g t g t g t c c t t t g t c g a t a c t g g t a c t a a t g c g g t t c g c g c g t t a a t t g g g a g t g a t t

4080 T3 promoter SNR52 promoter a g g g a a c a a a a g c t g g a g c t c c a c c g c g g a c a a t c t t t g a a a a g a t a a t g t a t g a t t a t g t c c c t t g t t t t c g a c c t c g a g g t g g c g c c t g t t a g a a a c t t t t c t a t t a c a t a c t a a t a c

4140 SNR52 promoter c t t t c a c t c a t a t t t a t a c a g a a a c t t g a t g t t t t c t t t c g a g t a t a t a c a a g g t g a t t a g a a a g t g a g t a t a a a t a t g t c t t t g a a c t a c a a a a g a a a g c t c a t a t a t g t t c c a c t a a t

BmtINheI 4200 SNR52 promoter c a t g t a c g t t t g a a g t a c a a c t c t a g a t t t t g t a g t g c c c t c t t g g g c t a g c g g t a a a g g g t a c a t g c a a a c t t c a t g t t g a g a t c t a a a a c a t c a c g g g a g a a c c c g a t c g c c a t t t c c

4260 SNR52 promoter t g c g c a t t t t t t c a c a c c c t a c a a t g t t c t g t t c a a a a g a t t t t g g t c a a a c g c t g t a g a a c g c g t a a a a a a g t g t g g g a t g t t a c a a g a c a a g t t t t c t a a a a c c a g t t t g c g a c a t c t

4320 SNR52 promoter a g t g a a a g t t g g t g c g c a t g t t t c g g c g t t c g a a a c t t c t c c g c a g t g a a a g a t a a a t g a t c a c t t t c a a c c a c g c g t a c a a a g c c g c a a g c t t t g a a g a g g c g t c a c t t t c t a t t t a c t

4380 SNR52 promoter gRNA scaffold t c a a g g a c c a a a t a g g c a a t g g g t t t t a g a g c t a g a a a t a g c a a g t t a a a a t a a g g c t a g a g t t c c t g g t t t a t c c g t t a c c c a a a a t c t c g a t c t t t a t c g t t c a a t t t t a t t c c g a t c

4440 gRNA scaffold SUP4 terminator t c c g t t a t c a a c t t g a a a a a g t g g c a c c g a g t c g g t g g t g c t t t t t t t g t t t t t t a t g t c a g g c a a t a g t t g a a c t t t t t c a c c g t g g c t c a g c c a c c a c g a a a a a a a c a a a a a a t a c a g

4500 SUP4 terminator t t c g a g t c a t g t a a t t a g t t a t a a g c a t g t g a g c t a a g a c a c t g t a a t t g c c a a t c t a a a a a g c t c a g t a c a t t a a t c a a t a t t c g t a c a c t c g a t t c t g t g a c a t t a a c g g t t a g a t t t

4560 ADH1 terminator c g a t a c c a c g g c c g c t c t a g a g a a a t g g g g a g c g a t t t g c a g g c a t t t g c t c g g c a t g c c g c t a t g g t g c c g g c g a g a t c t c t t t a c c c c t c g c t a a a c g t c c g t a a a c g a g c c g t a c g g

4620 ADH1 terminator g g t a g a g g t g t g g t c a a t a a g a g c g a c c t c a t g c t a t a c c t g a g a a a g c a a c c t g a c c t a c c a t c t c c a c a c c a g t t a t t c t c g c t g g a g t a c g a t a t g g a c t c t t t c g t t g g a c t g g a t

4680 ADH1 terminator G A G T C A C T T T A A A A pHybLex-R c a g g a a a g a g t t a c t c a a g a a t a a g a a t t t t c g t t t t a a a a c c t a a g a g t c a c t t t a a a a g t c c t t t c t c a a t g a g t t c t t a t t c t t a a a a g c a a a a t t t t g g a t t c t c a g t g a a a t t t t

4740 ADH1 terminator T T T G T A T A C A C pHybLex-R t t t g t a t a c a c t t a t t t t t t t t a t a a c t t a t t t a a t a a t a a a a a t c a t a a a t c a t a a g a a a a a c a t a t g t g a a t a a a a a a a a t a t t g a a t a a a t t a t t a t t t t t a g t a t t t a g t a t t c t t

4800 AG (in frame with SV40 NLS) *SGSSG 130 VKRKKKP ADH1 terminator SV40 NLS *SGSSGVKRKKKPAG a t t c g c c t c g a g a g t c t a g g a t c c g g a a c t a c c t a c c t t g c g c t t t t t c t t g g g a g c t c c t a a g c g g a g c t c t c a g a t c c t a g g c c t t g a t g g a t g g a a c g c g a a a a a g a a c c c t c g a g g

4860 (in frame with Cas9(N)) EDGGLQSLDIRTEYLGTISQ EDGGLQSLDIRTEYLGTISQ c t c a t c c c c t c c g a g c t g t g a g a g g t c a a t c c g a g t t t c g t a a a g t c c g g t g a t g c t c t g g a g t a g g g g a g g c t c g a c a c t c t c c a g t t a g g c t c a a a g c a t t t c a g g c c a c t a c g a g a c

4920 (in frame with Cas9(N)) HILTADLVEKTSTYRKRDIT HILTADLVEKTSTYRKRDIT g t g g a t g a g g g t g g c g t c c a g c a c t t c t t t g g t g g a g g t a t a g c g t t t c c g g t c g a t g g t c a c c t a c t c c c a c c g c a g g t c g t g a a g a a a c c a c c t c c a t a t c g c a a a g g c c a g c t a c c a

4980 (in frame with Cas9(N)) TDFYKFAAPAGLNTLTFLHI TDFYKFAAPAGLNTLTFLHI g g t g t c g a a g t a c t t g a a t g c g g c a g g g g c a c c a a g a t t g g t g a g g g t g a a c a g g t g g a t c c a c a g c t t c a t g a a c t t a c g c c g t c c c c g t g g t t c t a a c c a c t c c c a c t t g t c c a c c t a

5040 (in frame with Cas9(N)) INEAQERIPKDRHKNYASLV INEAQERIPKDRHKNYASLV g a t a t t c t c a g c c t g c t c c c g a a t t g g t t t g t c t c t g t g c t t a t t g t a t g c g c t a a g g a c c t a t a a g a g t c g g a c g a g g g c t t a a c c a a a c a g a g a c a c g a a t a a c a t a c g c g a t t c c t g

5100 (in frame with Cas9(N)) KDLNADALIVRKSFESIQEI KDLNADALIVRKSFESIQEI t t t a t c g a g g t t g g c a t c a g c g a g a a t g a c g c g t t t a g a a a a c t c g g a a a t c t g c t c a a t a a a t a g c t c c a a c c g t a g t c g c t c t t a c t g c g c a a a t c t t t t g a g c c t t t a g a c g a g t t a

5160 (in frame with Cas9(N)) IEDLYHKHQEVFLQKQENDE IEDLYHKHQEVFLQKQENDE g a t c t c g t c c a g a t a a t g c t t a t g c t g c t c c a c a a a c a g t t g c t t t t g c t c g t t g t c c t c c t a g a g c a g g t c t a t t a c g a a t a c g a c g a g g t g t t t g t c a a c g a a a a c g a g c a a c a g g a g

5220 (in frame with Cas9(N)) PSGKLKEYHSALYLFNVYKS PSGKLKEYHSALYLFNVYKS g g g g c t c c c c t t c a g c t t c t c g t a a t g g c t t g c a a g g t a g a g g a a a t t c a c g t a c t t g g a c c c c g a g g g g a a g t c g a a g a g c a t t a c c g a a c g t t c c a t c t c c t t t a a g t g c a t g a a c c t

5280 (in frame with Cas9(N)) PLALENGKQLEGASALMRKR PLALENGKQLEGASALMRKR g g g g a g a g c c a g c t c a t t a c c c t t c t g a a g t t c g c c a g c g g a g g c g a g c a t c c g c t t c c g c c c c t c t c g g t c g a g t a a t g g g a a g a c t t c a a g c g g t c g c c t c c g c t c g t a g g c g a a g g c

5340 (in frame with Cas9(N)) GNELEFLSYKPLKIILDKKV GNELEFLSYKPLKIILDKKV c c c a t t c t c c a g t t c g a a c a g g c t a t a c t t t g g g a g c t t g a t g a t a a g g t c c t t c t t g a c g g g t a a g a g g t c a a g c t t g t c c g a t a t g a a a c c c t c g a a c t a c t a t t c c a g g a a g a a c t g

5400 (in frame with Cas9(N)) EKYGKAELFDIPNKEFSSRE EKYGKAELFDIPNKEFSSRE t t c c t t g t a a c c t t t g g c t t c g a g a a a g t c a a t t g g g t t c t t c t c g a a g g a g c t g c g c t c a a g g a a c a t t g g a a a c c g a a g c t c t t t c a g t t a a c c c a a g a a g a g c t t c c t c g a c g c g a g

5460 (in frame with Cas9(N)) MITIGLLEKVSKLKKSKGKE MITIGLLEKVSKLKKSKGKE c a t a a t g g t g a t a c c c a g c a g t t c c t t c a c g c t c t t g a g c t t c t t a g a c t t t c c t t t t t c g t a t t a c c a c t a t g g g t c g t c a a g g a a g t g c g a g a a c t c g a a g a a t c t g a a a g g a a a a a g

5520 (in frame with Cas9(N)) VKAVVLVSYAVTPSDFGGYK VKAVVLVSYAVTPSDFGGYK c a c c t t a g c c a c g a c c a g g a c a g a a t a a g c c a c a g t t g g t g a a t c g a a t c c g c c g t a c t t g t g g a a t c g g t g c t g g t c c t g t c t t a t t c g g t g t c a a c c a c t t a g c t t a g g c g g c a t g a a

5580 (in frame with Cas9(N)) KPDWDKKRAILKDSNRKPLI KPDWDKKRAILKDSNRKPLI c t t a g g g t c c c a a t c c t t c t t c c t t g c a a t g a g c t t g t c g g a g t t g c g c t t g g g g a g g a t g a a t c c c a g g g t t a g g a a g a a g g a a c g t t a c t c g a a c a g c c t c a a c g c g a a c c c c t c c t a

5640 (in frame with Cas9(N)) SEKSFGGTQVETKKVINVQP SEKSFGGTQVETKKVINVQP t g a t t c c t t a g a a a a g c c t c c g g t t t g c a c c t c g g t t t t c t t c a c g a t g t t c a c t t g a g g a c t a a g g a a t c t t t t c g g a g g c c a a a c g t g g a g c c a a a a g a a g t g c t a c a a g t g a a c t c c

5700 (in frame with Cas9(N)) MSLVKRVTAFDRGKDWVIEG MSLVKRVTAFDRGKDWVIEG c a t a g a a a g c a c t t t g c g c a c g g t a g c g a a a t c t c t g c c c t t g t c c c a c a c g a t t t c g c c g t a t c t t t c g t g a a a c g c g t g c c a t c g c t t t a g a g a c g g g a a c a g g g t g t g c t a a a g c g g

5760 (in frame with Cas9(N)) TEGNTEILPRKRIEGNALTI TEGNTEILPRKRIEGNALTI a g t c t c c c c a t t a g t c t c g a t g a g c g g c c t c t t c c g g a t c t c a c c g t t t g c a a g g g t g a t t c a g a g g g g t a a t c a g a g c t a c t c g c c g g a g a a g g c c t a g a g t g g c a a a c g t t c c c a c t a

5820 (in frame with Cas9(N)) ETKFFNMINSYFFYKATAKG ETKFFNMINSYFFYKATAKG t t c g g t c t t g a a g a a a t t c a t g a t g t t g c t g t a g a a g a a g t a c t t t g c g g t g g c c t t t c c a a g c c a g a a c t t c t t t a a g t a c t a c a a c g a c a t c t t c t t c a t g a a a c g c c a c c g g a a a g g

5880 (in frame with Cas9(N)) IEQESKAIMKRVDYVKYDGY IEQESKAIMKRVDYVKYDGY g a t c t c c t g c t c a g a t t t g g c a a t c a t c t t g c g c a c g t c g t a g a c c t t g t a g t c t c c g t a c t a g a g g a c g a g t c t a a a c c g t t a g t a g a a c g c g t g c a g c a t c t g g a a c a t c a g a g g c a t

5940 (in frame with Cas9(N)) VFESELKPYKKILATGVVAN VFESELKPYKKILATGVVAN c a c g a a c t c g c t t t c c a g c t t t g g g t a c t t c t t g a t t a g g g c g g t t c c c a c c a c g g c a t t g t g c t t g a g c g a a a g g t c g a a a c c c a t g a a g a a c t a a t c c c g c c a a g g g t g g t g c c g t a a

6000 (in frame with Cas9(N)) LYADHAHHYNNIERVKYFQF LYADHAHHYNNIERVKYFQF c a g a t a t g c g t c a t g a g c a t g a t g a t a g t t a t t g a t c t c c c g c a c t t t g t a g a a t t g a a a g t c t a t a c g c a g t a c t c g t a c t a c t a t c a a t a a c t a g a g g g c g t g a a a c a t c t t a a c t t t

6060 (in frame with Cas9(N)) DKRFDSVLKSKLTIVKVERI DKRFDSVLKSKLTIVKVERI g t c c t t t c t g a a a t c g c t g a c c a g c t t g g a c t t c a g g g t a a t c a c c t t c a c c t c g c g g a t c a g g a a a g a c t t t a g c g a c t g g t c g a a c c t g a a g t c c c a t t a g t g g a a g t g g a g c g c c t a

6120 (in frame with Cas9(N)) LKDNEDYKTNMRSDLIQAVH LKDNEDYKTNMRSDLIQAVH a a g t t t g t c g t t t t c g t c g t a c t t g g t a t t c a t g c g t g a g t c a a g g a t t t g g g c t a c a t g t t c a a a c a g c a a a a g c a g c a t g a a c c a t a a g t a c g c a c t c a g t t c c t a a a c c c g a t g t a c

6180 (in frame with Cas9(N)) KTIQRTEVLQRKIFGAKDLE KTIQRTEVLQRKIFGAKDLE t t t a g t a a t c t g c c g a g t c t c c a c a a g t t g c c g t t t a a t g a a t c c t g c t t t g t c c a g t t c a a a t c a t t a g a c g g c t c a g a g g t g t t c a a c g g c a a a t t a c t t a g g a c g a a a c a g g t c a a g

6240 (in frame with Cas9(N)) SLGGREAKTLNDFKRQTILK SLGGREAKTLNDFKRQTILK g c t c a g t c c g c c c c t c t c a g c c t t a g t g a g a t t g t c g a a c t t c c g c t g a g t g a t t a g c t t c g a g t c a g g c g g g g a g a g t c g g a a t c a c t c t a a c a g c t t g a a g g c g a c t c a c t a a t c g a a

6300 (in frame with Cas9(N)) ANLLQRWYNKMKKVVEESPV ANLLQRWYNKMKKVVEESPV t g c g t t c a g a a g c t g g c g c c a a t a g t t c t t c a t c t t t t t c a c g a c t t c c t c t g a g g g c a c a c g c a a g t c t t c g a c c g c g g t t a t c a a g a a g t a g a a a a a g t g c t g a a g g a g a c t c c c g t g

6360 (in frame with Cas9(N)) NDSKGRNKDSRTLVKNDISD NDSKGRNKDSRTLVKNDISD g t t a t c t g a c t t t c c c c g g t t c t t g t c t g a g c g a g t c a g c a c c t t a t t g t c a a t t g a g t c c a a t a g a c t g a a a g g g g c c a a g a a c a g a c t c g c t c a g t c g t g g a a t a a c a g t t a a c t c a g

6420 (in frame with Cas9(N)) DKLFSQPVIHDVDYDSLRNI DKLFSQPVIHDVDYDSLRNI g t c t t t g a g g a a g c t c t g a g g g a c g a t g t g g t c c a c g t c g t a g t c t g a g a g c c t g t t g a t c a g a a a c t c c t t c g a g a c t c c c t g c t a c a c c a g g t g c a g c a t c a g a c t c t c g g a c a a c t a

6480 (in frame with Cas9(N)) DLEQDVYMDRGNQLYYLYLK DLEQDVYMDRGNQLYYLYLK a t c a a g c t c t t g g t c c a c a t a c a t a t c g c g t c c a t t t t g a a g g t a g t a c a g a t a g a g c t t t a g t t c g a g a a c c a g g t g t a t g t a t a g c g c a g g t a a a a c t t c c a t c a t g t c t a t c t c g a a

6540 (in frame with Cas9(N)) ENQLQTNEVPHEKLIQSGLE ENQLQTNEVPHEKLIQSGLE c t c a t t c t g a a g c t g g g t g t t t t c c a c c g g g t g c t c t t t a a g g a t c t g t g a c c c g a g c t c g a g t a a g a c t t c g a c c c a c a a a a g g t g g c c c a c g a g a a a t t c c t a g a c a c t g g g c t c g a g

6600 (in frame with Cas9(N)) KIGEEIRKMRERSNKQGKQT KIGEEIRKMRERSNKQGKQT c t t a a t c c c t t c c t c a a t t c t t t t c a t c c g c t c c c t g c t g t t c t t c t g t c c c t t c t g g g t g a a t t a g g g a a g g a g t t a a g a a a a g t a g g c g a g g g a c g a c a a g a a g a c a g g g a a g a c c c a

6660 (in frame with Cas9(N)) TQNERAMEIVINEPKHRGMV TQNERAMEIVINEPKHRGMV a g t c t g g t t t t c c c t g g c c a t c t c g a t g a c a a t g t t c t c t g g t t t a t g g c g a c c c a t c a c t c a g a c c a a a a g g g a c c g g t a g a g c t a c t g t t a c a a g a g a c c a a a t a c c g c t g g g t a g t g

6720 (in frame with Cas9(N)) KVLEDVVKVTQLIGKKIAPS KVLEDVVKVTQLIGKKIAPS c t t g a c c a g c t c g t c c a c g a c t t t c a c g g t c t g g a g a a t a c c c t t c t t a a t a g c g g g t g a g a a c t g g t c g a g c a g g t g c t g a a a g t g c c a g a c c t c t t a t g g g a a g a a t t a t c g c c c a c t

6780 (in frame with Cas9(N)) GALNAIHEHLSDGQGSVQAK GALNAIHEHLSDGQGSVQAK a c c a g c c a g a t t t g c g a t a t g c t c g t g c a g t g a g t c a c c t t g t c c a g a c a c t t g g g c c t t t g g t c g g t c t a a a c g c t a t a c g a g c a c g t c a c t c a g t g g a a c a g g t c t g t g a a c c c g g a a

6840 (in frame with Cas9(N)) QIDEKFTLSDDHILQMFNRN MDKLHEVAI QIDEKFTLSDDHILQMFNRN c t g g a t g t c c t c c t t a a a g g t c a g g g a a t c a t c a t g g a t a a g t t g c a t g a a g t t g c g a t t g a c c t a c a g g a g g a a t t t c c a g t c c c t t a g t a g t a c c t a t t c a a c g t a c t t c a a c g c t a a

6900 (in frame with Cas9(N)) AFGDSKLFDLITKGSQKDRI SESVALKEIKNGLSALFVPD AFGDSKLFDLITKGSQKDRI a g c g a a t c c g t c g c t c t t a a g g a a a t c a a g a a t g g t c t t t c c g c t c t g t t t g t c c c g g a t t c g c t t a g g c a g c g a g a a t t c c t t t a g t t c t t a c c a g a a a g g c g a g a c a a a c a g g g c c t a

6960 (in frame with Cas9(N)) GNILKRSLRGWGTYRRRKLQ SVDEFAAKASPSGISSPFKL GNILKRSLRGWGTYRRRKLQ t c c g t t g a t g a g t t t g c g g c t a a g g c g t c c c c a t c c g g t a t a t c t t c t c c g t t t a a g t t g a g g c a a c t a c t c a a a c g c c g a t t c c g c a g g g g t a g g c c a t a t a g a a g a g g c a a a t t c a a c

7020 (in frame with Cas9(N)) KMVKDDFLHAYTKLREEIME LHHFIVEQMGVGLKPLLDHF KMVKDDFLHAYTKLREEIME c t t c a t c a c t t t a t c g t c g a a c a g a t g g g c g t a g g t c t t a a g c c t c t c c t c g a t c a t t t c g a a g t a g t g a a a t a g c a g c t t g t c t a c c c g c a t c c a g a a t t c g g a g a g g a g c t a g t a a a g

7080 (in frame with Cas9(N)) RDEFLTLTLVIDELIDEKEE PVLEQSESQDNVLKDVLFLL RDEFLTLTLVIDELIDEKEE c c g g t c c t c g a a c a g a g t g a g a g t c a g g a c a a t g t c c t c a a g g a t g t c c t c t t t c t c c t c g g c c a g g a g c t t g t c t c a c t c t c a g t c c t g t t a c a g g a g t t c c t a c a g g a g a a a g a g g a g

7140 (in frame with Cas9(N)) NDLFDKDKIIKLLDHYTGLS IVQKILVLDDLKQIMVGTQA NDLFDKDKIIKLLDHYTGLS a t t g t c c a g a a a a t c c t t g t c c t t g a t g a t c t t a a g c a g a t c a t g g t a g g t a c c c a g g c t t a a c a g g t c t t t t a g g a a c a g g a a c t a c t a g a a t t c g t c t a g t a c c a t c c a t g g g t c c g a

7200 (in frame with Cas9(N)) ANFRDEVGSIEVSDFC 570 EIKK Cas9(N) CVEPIFDPANLHRVEAFNFL ANFRDEVGSIEVSDFCEIKK t g c g t t g a a c c g a t c t t c g a c c c c g c t a a t c t c c a c a g a g t c g a a g c a t t c a a t t t t c t t a c g c a a c t t g g c t a g a a g c t g g g g c g a t t a g a g g t g t c t c a g c t t c g t a a g t t a a a a g a a

7260 565 560 555 550 FYDEKLQKVTVKRNTKFLLD Cas9(N) EIVLLQLLHGHLSIGLEQKV FYDEKLQKVTVKRNTKFLLD g a a a t a g t c c t c c t t c a g c t g c t t c a c g g t c a c c t t t c t a t t g g t c t t g a a c a g a a g g t c c t t t a t c a g g a g g a a g t c g a c g a a g t g c c a g t g g a a a g a t a a c c a g a a c t t g t c t t c c a g

7320 545 540 535 530 VIAKKQEGSLFAPKRMGETV Cas9(N) HNCLLLFA* VIAKKQEGSLFAPKRMGETV c a c a a t t g c c t t c t t t t g t t c g c c t g a c a g g a a t g c g g g c t t c c t c a t c c c t t c g g t g a c g t g t t a a c g g a a g a a a a c a a g c g g a c t g t c c t t a c g c c c g a a g g a g t a g g g a a g c c a c t g

BsrGI 7380 525 520 515 510 YKVKTLENYVTFYEYLLSHK Cas9(N) YKVKTLENYVTFYEYLLSHK a t a t t t c a c c t t g g t c a g c t c g t t g t a c a c g g t a a a g t a c t c g t a g a g c a g g g a a t g t t t t a t a a a g t g g a a c c a g t c g a g c a a c a t g t g c c a t t t c a t g a g c a t c t c g t c c c t t a c a a a

7440 505 500 495 490 PLVKENPLNKDFNTMREIFS Cas9(N) PLVKENPLNKDFNTMREIFS c g g c a g g a c c t t c t c a t t g g g g a g g t t c t t a t c g a a g t t a g t c a t c c g t t c g a t g a a t g a g c c g t c c t g g a a g a g t a a c c c c t c c a a g a a t a g c t t c a a t c a g t a g g c a a g c t a c t t a c t

7500 485 480 475 470 QASAGKDVVEEFNWPTITEE Cas9(N) QASAGKDVVEEFNWPTITEE c t g a g c g c t g g c t c c c t t g t c c a c c a c c t c t t c g a a g t t c c a g g g a g t g a t a g t t t c t t c g a c t c g c g a c c g a g g g a a c a g g t g g t g g a g a a g c t t c a a g g t c c c t c a c t a t c a a a g a a g

7560 465 460 455 450 SKRTMWAFRSNGRALPGVYY Cas9(N) SKRTMWAFRSNGRALPGVYY t g a c t t t c t g g t c a t c c a a g c a a a c c g g c t a t t t c c t c t g g c g a g g g g t c c c a c g t a g t a a c t g a a a g a c c a g t a g g t t c g t t t g g c c g a t a a a g g a g a c c g c t c c c c a g g g t g c a t c a t

7620 445 440 435 430 PIRFTLIKEIKERNDKLFPY Cas9(N) PIRFTLIKEIKERNDKLFPY g g g g a t g c g g a a a g t g a g a a t t t t c t c a a t c t t c t c c c t g t t g t c c t t a a g a a a g g g g t a c c c c t a c g c c t t t c a c t c t t a a a a g a g t t a g a a g a g g g a c a a c a g g a a t t c t t t c c c c a t

7680 425 420 415 410 FDEQRRLIAHLEGLHIQHPI Cas9(N) FDEQRRLIAHLEGLHIQHPI g a a g t c c t c t t g g c g c c g a a g g a t g g c g t g a a g c t c c c c a a g a t g g a t c t g g t g g g g a a t c t t c a g g a g a a c c g c g g c t t c c t a c c g c a c t t c g a g g g g t t c t a c c t a g a c c a c c c c t t a

7740 405 400 395 390 SGNDFTRQKRLLDERNLKVL Cas9(N) SGNDFTRQKRLLDERNLKVL g c t c c c a t t g t c g a a a g t g c g c t g c t t c c g c a g c a g a t c c t c c c g a t t c a g t t t c a c g a g c g a g g g t a a c a g c t t t c a c g c g a c g a a g g c g t c g t c t a g g a g g g c t a a g t c a a a g t g c t c

7800 385 380 375 370 LEETGDMKELIPKIFKYFEE Cas9(N) LEETGDMKELIPKIFKYFEE g a g c t c t t c g g t g c c g t c c a t c t t c t c a a g g a t a g g t t t g a t a a a c t t a t a a a a t t c c t c c t c g a g a a g c c a c g g c a g g t a g a a g a g t t c c t a t c c a a a c t a t t t g a a t a t t t t a a g g a g

7860 365 360 355 350 QSAGGDIYGAYGNKSQDFFI Cas9(N) QSAGGDIYGAYGNKSQDFFI t t g g g a g g c a c c t c c g t c g a t g t a g c c g g c g t a t c c a t t c t t t g a c t g g t c a a a g a a g a t a a c c c t c c g t g g a g g c a g c t a c a t c g g c c g c a t a g g t a a g a a a c t g a c c a g t t t c t t c t a

7920 345 340 335 330 EKYKEPLQQRVLAKLLTLDQ Cas9(N) EKYKEPLQQRVLAKLLTLDQ t t c c t t g t a c t t c t c c g g a a g c t g c t g c c g g a c g a g t g c c t t a a g c a g g g t c a g g t c c t g a a g g a a c a t g a a g a g g c c t t c g a c g a c g g c c t g c t c a c g g a a t t c g t c c c a g t c c a g g a c

7980 325 320 315 310 HHEDYRKIMSASLPAKTIET Cas9(N) HHEDYRKIMSASLPAKTIET g t g g t g c t c a t c a t a c c g c t t a a t c a t t g a a g c t g a a a g a g g g g c c t t g g t g a t t t c a g t c a c c a c g a g t a g t a t g g c g a a t t a g t a a c t t c g a c t t t c t c c c c g g a a c c a c t a a a g t c a

8040 305 300 295 290 NVRLIDSLLIADSLNKAALF Cas9(N) NVRLIDSLLIADSLNKAALF g t t c a c g c g g a g a a t g t c a g a c a g c a g g a t g g c g t c a g a a a g a t t c t t a g c a g c a a g g a a c a a g t g c g c c t c t t a c a g t c t g t c g t c c t a c c g c a g t c t t t c t a a g a a t c g t c g t t c c t t

8100 285 280 275 270 LDAYQDGIQALLNDLDDDYT Cas9(N) LDAYQDGIQALLNDLDDDYT a a g g t c g g c g t a t t g g t c a c c g a t c t g g g c c a g g a g g t t g t c g a g a t c g t c a t c a t a g g t t t c c a g c c g c a t a a c c a g t g g c t a g a c c c g g t c c t c c a a c a g c t c t a g c a g t a g t a t c c a

8160 265 260 255 250 DKSLQLKADEALDFNSKFNP Cas9(N) DKSLQLKADEALDFNSKFNP g t c c t t g g a c a g t t g c a g c t t g g c a t c c t c t g c c a g g t c g a a a t t a g a c t t g a a a t t g g g c a g g a a c c t g t c a a c g t c g a a c c g t a g g a g a c g g t c c a g c t t t a a t c t g a a c t t t a a c c c

8220 245 240 235 230 TLGLSLAILNGFLGNKKEGP Cas9(N) TLGLSLAILNGFLGNKKEGP a g t c a g c c c a a g t g a a a g a g c a a t c a g g t t c c c g a a c a g t c c a t t t t t c t t t t c a c c c g g t c a g t c g g g t t c a c t t t c t c g t t a g t c c a a g g g c t t g t c a g g t a a a a a g a a a a g t g g g c c

BlpI 8280 225 220 215 210 LQAILNELRRSKSLRASLIA Cas9(N) LQAILNELRRSKSLRASLIA c a g c t g a g c g a t a a g a t t c t c a a g t c t g c g g c t c t t t g a c a g c c g g g c t g a c a g a a t g g c g t c g a c t c g c t a t t c t a a g a g t t c a g a c g c c g a g a a a c t g t c g g c c c g a c t g t c t t a c c g

8340 205 200 195 190 KADVGSANIPNEEFLQNYTQ Cas9(N) KADVGSANIPNEEFLQNYTQ c t t g g c a t c g a c t c c g c t t g c a t t g a t g g g g t t c t c t t c a a a g a g c t g g t t g t a a g t c t g g a a c c g t a g c t g a g g c g a a c g t a a c t a c c c c a a g a g a a g t t t c t c g a c c a a c a t t c a g a c

8400 185 180 175 170 VLQIFLKDVDSNDPNLDGEI Cas9(N) VLQIFLKDVDSNDPNLDGEI c a c c a g c t g g a t g a a c a g t t t a t c c a c a t c g c t a t t g t c a g g a t t g a g g t c g c c c t c g a t g t g g t c g a c c t a c t t g t c a a a t a g g t g t a g c g a t a a c a g t c c t a a c t c c a g c g g g a g c t a

8460 165 160 155 150 LFHGRFKIMHALALYILRLD Cas9(N) LFHGRFKIMHALALYILRLD c a g g a a g t g g c c t c t g a a c t t a a t c a t g t g g g c c a g g g c c a g g t a g a t a a g c c g g a g g t c g t c c t t c a c c g g a g a c t t g a a t t a g t a c a c c c g g t c c c g g t c c a t c t a t t c g g c c t c c a g

8520 145 140 135 130 AKDTSDVLKKRLHYITPYKE Cas9(N) AKDTSDVLKKRLHYITPYKE t g c c t t g t c g g t t g a g t c c a c c a g c t t t t t g c g a a g a t g g t a g a t g g t t g g g t a t t t c t c a c g g a a c a g c c a a c t c a g g t g g t c g a a a a a c g c t t c t a c c a t c t a c c a a c c c a t a a a g a g

SgrDI 8580 125 120 115 110 HYAVEDVINGFIPHREHKKD Cas9(N) HYAVEDVINGFIPHREHKKD g t g a t a g g c c a c c t c g t c g a c g a t a t t a c c g a a g a t g g g a t g c c g t t c a t g c t t c t t a t c c a c t a t c c g g t g g a g c a g c t g c t a t a a t g g c t t c t a c c c t a c g g c a a g t a c g a a g a a t a g

8640 105 100 95 90 EEVLFSEELRHFFSDDVKAM Cas9(N) EEVLFSEELRHFFSDDVKAM c t c t t c c a c g a g g a a t g a t t c t t c c a g c c t a t g g a a g a a g c t g t c g t c g a c c t t t g c c a t g a g a a g g t g c t c c t t a c t a a g a a g g t c g g a t a c c t t c t t c g a c a g c a g c t g g a a a c g g t a

8700 85 80 75 70 ENSFIEQLYCIRNKRRTYRR Cas9(N) ENSFIEQLYCIRNKRRTYRR t t c g t t g g a g a a t a t c t c t t g a a g g t a a c a a a t c c g g t t c t t c c t c c t g g t g t a c c g c c g a a g c a a c c t c t t a t a g a g a a c t t c c a t t g t t t a g g c c a a g a a g g a g g a c c a c a t g g c g g c

BsaI 8760 65 60 55 50 RATRKLRTAEATEGSDFLLA Cas9(N) RATRKLRTAEATEGSDFLLA t c t a g c a g t c c g c t t g a g g c g g g t t g c t t c a g c g g t c t c c c c t g a a t c a a a g a g g a g a g c a g a t c g t c a g g c g a a c t c c g c c c a a c g a a g t c g c c a g a g g g g a c t t a g t t t c t c c t c t c g

8820 45 40 35 30 GILNKKISHRDTNGLVKFKK Cas9(N) GILNKKISHRDTNGLVKFKK t c c g a t c a g a t t t t t c t t g a t g g a a t g c c g g t c g g t g t t c c c g a g c a c c t t g a a c t t c t t a g g c t a g t c t a a a a a g a a c t a c c t t a c g g c c a g c c a c a a g g g c t c g t g g a a c t t g a a g a a

8880 25 20 15 10 SPVKYEDTIVAWGVSNTGID Cas9(N) SPVKYEDTIVAWGVSNTGID a g a g g g c a c c t t g t a c t c g t c a g t g a t g a c g g c c c a c c c g a c g c t a t t a g t c c c g a t g t c t c t c c c g t g g a a c a t g a g c a g t c a c t a c t g c c g g g t g g g c t g c g a t a a t c a g g g c t a c a g

8940 5 1 LGISYKKDM Cas9(N) Kozak sequence LGISYKKDM c a g t c c g a t a g a a t a c t t c t t g t c c a t g g t g g c g a g a t c t a a t t c g a t c c a t a t c a g a t c g t c a g g c t a t c t t a t g a a g a a c a g g t a c c a c c g c t c t a g a t t a a g c t a g g t a t a g t c t a g

9000 GAP promoter t t t t g t t t g t t t a t g t g t g t t t a t t c g a a a c t a a g t t c t t g g t g t t t t a a a a c t a a a a a a a a a a c a a a c a a a t a c a c a c a a a t a a g c t t t g a t t c a a g a a c c a c a a a a t t t t g a t t t t t t

9060 GAP promoter G T C T T A A T G T T A GPDpro-F a a g a c t a a c t a t a a a a g t a g a a t t t a a g a a g t t t a a g a a a t a g a t t t a c a g a a t t a c a a t t t c t g a t t g a t a t t t t c a t c t t a a a t t c t t c a a a t t c t t t a t c t a a a t g t c t t a a t g t t a

9120 GAP promoter G T T A T G G A T G G C GPDpro-F c a a t a c c t a c c g t c t t t a t a t a c t t a t t a g t c a a g t a g g g g a a t a a t t t c a g g g a a c t g g g t t a t g g a t g g c a g a a a t a t a t g a a t a a t c a g t t c a t c c c c t t a t t a a a g t c c c t t g a c c

9180 GAP promoter t t t c a a c c t t t t t t t t c a g c t t t t t c c a a a t c a g a g a g a g c a g a a g g t a a t a g a a g g t g t a a a g t t g g a a a a a a a a g t c g a a a a a g g t t t a g t c t c t c t c g t c t t c c a t t a t c t t c c a c a

9240 GAP promoter a a g a a a a t g a g a t a g a t a c a t g c g t g g g t c a a t t g c c t t g t g t c a t c a t t t a c t c c a g g c t t c t t t t a c t c t a t c t a t g t a c g c a c c c a g t t a a c g g a a c a c a g t a g t a a a t g a g g t c c g

9300 GAP promoter a g g t t g c a t c a c t c c a t t g a g g t t g t g c c c g t t t t t t g c c t g t t t g t g c c c c t g t t c t c t t c c a a c g t a g t g a g g t a a c t c c a a c a c g g g c a a a a a a c g g a c a a a c a c g g g g a c a a g a g a

9360 GAP promoter g t a g t t g c g c t a a g a g a a t g g a c c t a t g a a c t g a t g g t t g g t g a a g a a a a c a a t a t t t t g c a t c a a c g c g a t t c t c t t a c c t g g a t a c t t g a c t a c c a a c c a c t t c t t t t g t t a t a a a a c

9420 GAP promoter g t g c t g g g a t t c t t t t t t t t t c t g g a t g c c a g c t t a a a a a g c g g g c t c c a t t a t a t t t a g c a c g a c c c t a a g a a a a a a a a a g a c c t a c g g t c g a a t t t t t c g c c c g a g g t a a t a t a a a t c

9480 GAP promoter t g g a t g c c a g g a a t a a a c t g t t c a c c c a g a c a c c t a c g a t g t t a t a t a t t c t g t g t a a c c a c c t a c g g t c c t t a t t t g a c a a g t g g g t c t g t g g a t g c t a c a a t a t a t a a g a c a c a t t g g

9540 GAP promoter c g c c c c c t a t t t t g g g c a t g t a c g g g t t a c a g c a g a a t t a a a a g g c t a a t t t t t t g a c t a g c g g g g g a t a a a a c c c g t a c a t g c c c a a t g t c g t c t t a a t t t t c c g a t t a a a a a a c t g a t

9600 GAP promoter a a t a a a g t t a g g a a a a t c a c t a c t a t t a a t t a t t t a c g t a t t c t t t g a a a t g g c a g t a t t t t a t t t c a a t c c t t t t a g t g a t g a t a a t t a a t a a a t g c a t a a g a a a c t t t a c c g t c a t a a

9660 GAP promoter KS primer C T A T G G C pBluescriptKS g a t a a t g a t a a a c t c g a a c t g a c t a g t a g a c t g a a t t c g a t a t c a a g c t t a t c g a t a c c g c t a t t a c t a t t t g a g c t t g a c t g a t c a t c t g a c t t a a g c t a t a g t t c g a a t a g c t a t g g c

PspOMIApaIPspXIAbsI 9720 KS primer T3 promoter A G C T G G A G C T pBluescriptKS t c g a c c t c g a g g g g g g g c c c g g t a c c a g c t t t t g t t c c c t t t a g t g a g g g t t a a t t t c g a a g c t g g a g c t c c c c c c c g g g c c a t g g t c g a a a a c a a g g g a a a t c a c t c c c a a t t a a a g c t

9780 M13 rev lac operator g c t t g g c g t a a t c a t g g t c a t a g c t g t t t c c t g t g t g a a a t t g t t a t c c g c t c a c a a t t c c g a a c c g c a t t a g t a c c a g t a t c g a c a a a g g a c a c a c t t t a a c a a t a g g c g a g t g t t a a g

9840 -10 -35 lac promoter c a c a c a a c a t a c g a g c c g g a a g c a t a a a g t g t a a a g c c t g g g g t g c c t a a t g a g t g a g g t g t g t g t t g t a t g c t c g g c c t t c g t a t t t c a c a t t t c g g a c c c c a c g g a t t a c t c a c t c c a

9900 RB T-DNA repeat a c g g a t c g a t c c t g a c a g g a t a t a t t g g c g g g t a a a c c t a a g a g a a a a g a g c g t t t a t t a t g c c t a g c t a g g a c t g t c c t a t a t a a c c g c c c a t t t g g a t t c t c t t t t c t c g c a a a t a a t

9960 g a a t a a t c g g a t a t t t a a a a g g g c g t g a a a a g g t t t a t c c g t t c g t c c a t t t g t a t g t g c c t t a t t a g c c t a t a a a t t t t c c c g c a c t t t t c c a a a t a g g c a a g c a g g t a a a c a t a c a c g

10,020 a t g c c a a c c a c a g g g t t c c c c a g a t c t g g c g c c g g c c a g c g a g a c g a g c a a g a t t g g c c g t a c g g t t g g t g t c c c a a g g g g t c t a g a c c g c g g c c g g t c g c t c t g c t c g t t c t a a c c g g c

AgeI 10,080 c g g t c a c a c t g c t t c c g g t a g t c a a t a a a c c g g t a a a c c a g c a a t a g a c a t a a g c g g c t a g c c a g t g t g a c g a a g g c c a t c a g t t a t t t g g c c a t t t g g t c g t t a t c t g t a t t c g c c g a t

10,140 t t t a a c g a c c c t g c c c t g a a c c g a c g a c c g g g t c g a a t t t g c t t t c g a a c c c c a g a g t c c a a a t t g c t g g g a c g g g a c t t g g c t g c t g g c c c a g c t t a a a c g a a a g c t t g g g g t c t c a g g

10,200 265 260 255 250 *FFEDLLRYFAIRQSDPAA NeoR/KanR *FFEDLLRYFAIRQSDPAA c g c t c a g a a g a a c t c g t c a a g a a g g c g a t a g a a g g c g a t g c g c t g c g a a t c g g g a g c g g c g c g a g t c t t c t t g a g c a g t t c t t c c g c t a t c t t c c g c t a c g c g a c g c t t a g c c c t c g c c g

10,260 245 240 235 230 IGYLVLFRDAWEGGLEEAID NeoR/KanR T T A T A G Neo-F IGYLVLFRDAWEGGLEEAID g a t a c c g t a a a g c a c g a g g a a g c g g t c a g c c c a t t c g c c g c c a a g c t c t t c a g c a a t a t c c t a t g g c a t t t c g t g c t c c t t c g c c a g t c g g g t a a g c g g c g g t t c g a g a a g t c g t t a t a g

RsrII 10,320 225 220 215 210 RTALAIDQYRDAVGLRGCDI NeoR/KanR T G C C C A T C G G T T G C Neo-F RTALAIDQYRDAVGLRGCDI a c g g g t a g c c a a c g c t a t g t c c t g a t a g c g g t c c g c c a c a c c c a g c c g g c c a c a g t c g a t t g c c c a t c g g t t g c g a t a c a g g a c t a t c g c c a g g c g g t g t g g g t c g g c c g g t g t c a g c t a

10,380 205 200 195 190 FGSFRGNEVMINPLCADGHT NeoR/KanR FGSFRGNEVMINPLCADGHT g a a t c c a g a a a a g c g g c c a t t t t c c a c c a t g a t a t t c g g c a a g c a g g c a t c g c c a t g g g t c t t a g g t c t t t t c g c c g g t a a a a g g t g g t a c t a t a a g c c g t t c g t c c g t a g c g g t a c c c a

10,440 185 180 175 170 VVLDEGDPMRAKLRAFLEAP NeoR/KanR VVLDEGDPMRAKLRAFLEAP c a c g a c g a g a t c c t c g c c g t c g g g c a t g c g c g c c t t g a g c c t g g c g a a c a g t t c g g c t g g g t g c t g c t c t a g g a g c g g c a g c c c g t a c g c g c g g a a c t c g g a c c g c t t g t c a a g c c g a c c

10,500 165 160 155 150 ALGQHEEDLDDQDVLGAEMR NeoR/KanR ALGQHEEDLDDQDVLGAEMR c g c g a g c c c c t g a t g c t c t t c g t c c a g a t c a t c c t g a t c g a c a a g a c c g g c t t c c a t c c g g c g c t c g g g g a c t a c g a g a a g c a g g t c t a g t a g g a c t a g c t g t t c t g g c c g a a g g t a g g c

10,560 145 140 135 130 TRAREIRHKAQHDFPCTAPD NeoR/KanR TRAREIRHKAQHDFPCTAPD a g t a c g t g c t c g c t c g a t g c g a t g t t t c g c t t g g t g g t c g a a t g g g c a g g t a g c c g g a t c t c a t g c a c g a g c g a g c t a c g c t a c a a a g c g a a c c a c c a g c t t a c c c g t c c a t c g g c c t a g

10,620 125 120 115 110 LTHLRRMADAMISVKEAPAL NeoR/KanR LTHLRRMADAMISVKEAPAL a a g c g t a t g c a g c c g c c g c a t t g c a t c a g c c a t g a t g g a t a c t t t c t c g g c a g g a g c a a g t t c g c a t a c g t c g g c g g c g t a a c g t a g t c g g t a c t a c c t a t g a a a g a g c c g t c c t c g t t c

10,680 105 100 95 90 HSSLLDQGPVEGLLLWDRGA NeoR/KanR HSSLLDQGPVEGLLLWDRGA g t g a g a t g a c a g g a g a t c c t g c c c c g g c a c t t c g c c c a a t a g c a g c c a g t c c c t t c c c g c c a c t c t a c t g t c c t c t a g g a c g g g g c c g t g a a g c g g g t t a t c g t c g g t c a g g g a a g g g c g

10,740 85 80 75 70 ETVVDLVAACPVGTTALWSL NeoR/KanR ETVVDLVAACPVGTTALWSL t t c a g t g a c a a c g t c g a g c a c a g c t g c g c a a g g a a c g c c c g t c g t g g c c a g c c a c g a t a g a a g t c a c t g t t g c a g c t c g t g t c g a c g c g t t c c t t g c g g g c a g c a c c g g t c g g t g c t a t c

10,800 65 60 55 50 RAAEDQLENLAGSLDTKVFL NeoR/KanR RAAEDQLENLAGSLDTKVFL c c g c g c t g c c t c g t c c t g c a g t t c a t t c a g g g c a c c g g a c a g g t c g g t c t t g a c a a a a a g g g c g c g a c g g a g c a g g a c g t c a a g t a a g t c c c g t g g c c t g t c c a g c c a g a a c t g t t t t t c

10,860 45 40 35 30 VPRGQASLRFVAADSCGITQ NeoR/KanR VPRGQASLRFVAADSCGITQ a a c c g g g c g c c c c t g c g c t g a c a g c c g g a a c a c g g c g g c a t c a g a g c a g c c g a t t g t c t g t t g g c c c g c g g g g a c g c g a c t g t c g g c c t t g t g c c g c c g t a g t c t c g t c g g c t a a c a g a c

10,920 25 20 15 10 QAWDYGFLREVWAAPSGAHL NeoR/KanR G C C C A G T C A T A G C C G A A T A G Neo-R QAWDYGFLREVWAAPSGAHL t t g t g c c c a g t c a t a g c c g a a t a g c c t c t c c a c c c a a g c g g c c g g a g a a c c t g c g t g c a a a a c a c g g g t c a g t a t c g g c t t a t c g g a g a g g t g g g t t c g c c g g c c t c t t g g a c g c a c g t t

10,980 5 1 GDQEIM NeoR/KanR GDQEIM t c c a t c t t g t t c a a t c a t g c g a a a c g a t c c t c a t c c t g t c t c t t g a t c a g a t c t t g a t c c a g g t a g a a c a a g t t a g t a c g c t t t g c t a g g a g t a g g a c a g a g a a c t a g t c t a g a a c t a g g

11,040 c c t g c g c c a t c a g a t c c t t g g c g g c a a g a a a g c c a t c c a g t t t a c t t t g c a g g g c t t c c c g g a c g c g g t a g t c t a g g a a c c g c c g t t c t t t c g g t a g g t c a a a t g a a a c g t c c c g a a g g g

11,100 *WLAGWSAIGTRKSDMFGG a a c c t t a c c a g a g g g c g c c c c a g c t g g c a a t t c c g g t t c g c t t g c t g t c c a t a a a a c c g c t t g g a a t g g t c t c c c g c g g g g t c g a c c g t t a a g g c c a a g c g a a c g a c a g g t a t t t t g g c g

11,160 LRAIAMYAWQRSVLQHGVPE c c a g t c t a g c t a t c g c c a t g t a a g c c c a c t g c c t c g a g a c t a g t t g g t g t c c a a c c g g c t g g t c a g a t c g a t a g c g g t a c a t t c g g g t g a c g g a g c t c t g a t c a a c c a c a g g t t g g c c g a

11,220 VPAACPPAEPPGSLAQTYES c g a c g g g g g c a g c g c a a g g c g g t g c c t c c g g c g g g c c a c t c a a t g c t t g a g t a t a c t c a c g c t g c c c c c g t c g c g t t c c g c c a c g g a g g c c g c c c g g t g a g t t a c g a a c t c a t a t g a g t g

11,280 SKAECLRSRRRRSRRGVPKS t a g a c t t t g c t t c g c a a a g t c g t g a c c g c c t a c g g c g g c t g c g g c g c c c t a c g g g c t t g c a t c t g a a a c g a a g c g t t t c a g c a c t g g c g g a t g c c g c c g a c g c c g c g g g a t g c c c g a a c g

11,340 EPSRGARDSRERALGHIHPR t c t c c g g g c t t c g c c c t g c g c g g t c g c t g c g c t c c c t t g c c a g c c c g t g g a t a t g t g g a c a g a g g c c c g a a g c g g g a c g c g c c a g c g a c g c g a g g g a a c g g t c g g g c a c c t a t a c a c c t g

11,400 pBBR1 oriV HGRAAVPQSAESPGHVVRSS g a t g g c c g c g a g c g g c c a c c g g c t g g c t c g c t t c g c t c g g c c c g t g g a c a a c c c t g c t g g c t a c c g g c g c t c g c c g g t g g c c g a c c g a g c g a a g c g a g c c g g g c a c c t g t t g g g a c g a c c

11,460 pBBR1 oriV LSISLSRRGVLKVVPIAWRS a c a a g c t g a t g g a c a g g c t g c g c c t g c c c a c g a g c t t g a c c a c a g g g a t t g c c c a c c g g c t g t t c g a c t a c c t g t c c g a c g c g g a c g g g t g c t c g a a c t g g t g t c c c t a a c g g g t g g c c g

11,520 pBBR1 oriV GLRRGCVWRSWSRPRRGQGM t a c c c a g c c t t c g a c c a c a t a c c c a c c g g c t c c a a c t g c g c g g c c t g c g g c c t t g c c c c a a t g g g t c g g a a g c t g g t g t a t g g g t g g c c g a g g t t g a c g c g c c g g a c g c c g g a a c g g g g t

11,580 pBBR1 oriV t c a a t t t t t t t a a t t t t c t c t g g g g a a a a g c c t c c g g c c t g c g g c c t g c g c g c t t c g c t t a g t t a a a a a a a t t a a a a g a g a c c c c t t t t c g g a g g c c g g a c g c c g g a c g c g c g a a g c g a a

11,640 pBBR1 oriV g c c g g t t g g a c a c c a a g t g g a a g g c g g g t c a a g g c t c g c g c a g c g a c c g c g c a g c g g c t t c g g c c a a c c t g t g g t t c a c c t t c c g c c c a g t t c c g a g c g c g t c g c t g g c g c g t c g c c g a a

11,700 pBBR1 oriV g g c c t t g a c g c g c c t g g a a c g a c c c a a g c c t a t g c g a g t g g g g g c a g t c g a a g g g c g a a g c c g g a a c t g c g c g g a c c t t g c t g g g t t c g g a t a c g c t c a c c c c c g t c a g c t t c c c g c t t c

11,760 pBBR1 oriV c c c g c c c g c c t g c c c c c c g a g c c t c a c g g c g g c g a g t g c g g g g g t t c c a a g g g g g c a g c g g g g c g g g c g g a c g g g g g g c t c g g a g t g c c g c c g c t c a c g c c c c c a a g g t t c c c c c g t c g c

11,820 pBBR1 oriV c c a c c t t g g g c a a g g c c g a a g g c c g c g c a g t c g a t c a a c a a g c c c c g g a g g g g c c a c t t t g g t g g a a c c c g t t c c g g c t t c c g g c g c g t c a g c t a g t t g t t c g g g g c c t c c c c g g t g a a a

11,880 pBBR1 oriV t t g c c g g a g g g g g a g c c g c g c c g a a g g c g t g g g g g a a c c c c g c a g g g g t g c c c t t c t t t g a a c g g c c t c c c c c t c g g c g c g g c t t c c g c a c c c c c t t g g g g c g t c c c c a c g g g a a g a a a c

11,940 pBBR1 oriV *F g g c a c c a a a g a a c t a g a t a t a g g g c g a a a t g c g a a a g a c t t a a a a a t c a a c a a c t t a a a a c c g t g g t t t c t t g a t c t a t a t c c c g c t t t a c g c t t t c t g a a t t t t t a g t t g t t g a a t t t t

12,000 pBBR1 oriV LPTRLLENRCGGCYSMAYTL a a g g g g g g t a c g c a a c a g c t c a t t g c g g c a c c c c c c g c a a t a g c t c a t t g c g t a g g t t a a t t c c c c c c a t g c g t t g t c g a g t a a c g c c g t g g g g g g c g t t a t c g a g t a a c g c a t c c a a t t

12,060 pBBR1 oriV SFRYNVAVKVCRMITTASGF a g a a a a t c t g t a a t t g a c t g c c a c t t t t a c g c a a c g c a t a a t t g t t g t c g c g c t g c c g a a t c t t t t a g a c a t t a a c t g a c g g t g a a a a t g c g t t g c g t a t t a a c a a c a g c g c g a c g g c t t

12,120 pBBR1 oriV MEI LQLQNRMTGCGHPVGVAHLY a a g t t g c a g c t g a t t g c g c a t g g t g c c g c a a c c g t g c g g c a c c c c t a c c g c a t g g a g a t a t t c a a c g t c g a c t a a c g c g t a c c a c g g c g t t g g c a c g c c g t g g g g a t g g c g t a c c t c t a t

12,180 1 5 10 15 MATQSREIGIQAKNKPGHW pBBR1 oriV pBBR1 Rep SMATQSREIGIQAKNKPGHW AHGRLGSFDANLGLVLGTVP a g c a t g g c c a c g c a g t c c a g a g a a a t c g g c a t t c a a g c c a a g a a c a a g c c c g g t c a c t g g t c g t a c c g g t g c g t c a g g t c t c t t t a g c c g t a a g t t c g g t t c t t g t t c g g g c c a g t g a c c

12,240 20 25 30 35 VQTERKAHEAWAGLIARKPT pBBR1 Rep VQTERKAHEAWAGLIARKPT HLRFAFRMLRPGPKNRPFGR g t g c a a a c g g a a c g c a a a g c g c a t g a g g c g t g g g c c g g g c t t a t t g c g a g g a a a c c c a c g c a c g t t t g c c t t g c g t t t c g c g t a c t c c g c a c c c g g c c c g a a t a a c g c t c c t t t g g g t g c

12,300 40 45 50 55 AAMLLHHLVAQMGHQNAVVV pBBR1 Rep AAMLLHHLVAQMGHQNAVVV RCHQQMVEHRLHAVLVGHHD g c g g c a a t g c t g c t g c a t c a c c t c g t g g c g c a g a t g g g c c a c c a g a a c g c c g t g g t g g t c c g c c g t t a c g a c g a c g t a g t g g a g c a c c g c g t c t a c c c g g t g g t c t t g c g g c a c c a c c a g

12,360 60 65 70 75 SQKTLSKLIGRSLRTVQYAV pBBR1 Rep SQKTLSKLIGRSLRTVQYAV ALLCKGLEDSTRQPRDLVCD a g c c a g a a g a c a c t t t c c a a g c t c a t c g g a c g t t c t t t g c g g a c g g t c c a a t a c g c a g t c t c g g t c t t c t g t g a a a g g t t c g a g t a g c c t g c a a g a a a c g c c t g c c a g g t t a t g c g t c a g

12,420 80 85 90 95 KDLVAERWISVVKLNGPGTV pBBR1 Rep KDLVAERWISVVKLNGPGTV LVQHGLAPDGDHLEVAGAGH a a g g a c t t g g t g g c c g a g c g c t g g a t c t c c g t c g t g a a g c t c a a c g g c c c c g g c a c c g t g t t c c t g a a c c a c c g g c t c g c g a c c t a g a g g c a g c a c t t c g a g t t g c c g g g g c c g t g g c a c

12,480 100 105 110 115 SAYVVNDRVAWGQPRDQLRL pBBR1 Rep SAYVVNDRVAWGQPRDQLRL RGVHDIVAHRPALGAVLQAQ t c g g c c t a c g t g g t c a a t g a c c g c g t g g c g t g g g g c c a g c c c c g c g a c c a g t t g c g c c t g a g c c g g a t g c a c c a g t t a c t g g c g c a c c g c a c c c c g g t c g g g g c g c t g g t c a a c g c g g a c

12,540 120 125 130 135 SVFSAAVVVDHDDQDESLLG pBBR1 Rep SVFSAAVVVDHDDQDESLLG RHETGGHHNIVVVLVFRQQP t c g g t g t t c a g t g c c g c c g t g g t g g t t g a t c a c g a c g a c c a g g a c g a a t c g c t g t t g g g g a g c c a c a a g t c a c g g c g g c a c c a c c a a c t a g t g c t g c t g g t c c t g c t t a g c g a c a a c c c c

12,600 140 145 150 155 HGDLRRIPTLYPGEQQLPTG pBBR1 Rep HGDLRRIPTLYPGEQQLPTG M c a t g g c g a c c t g c g c c g c a t c c c g a c c c t g t a t c c g g g c g a g c a g c a a c t a c c g a c c g g c g t a c c g c t g g a c g c g g c g t a g g g c t g g g a c a t a g g c c c g c t c g t c g t t g a t g g c t g g c c g

12,660 160 165 170 175 PGEEPPSQPGIPGMEPDLPA pBBR1 Rep PGEEPPSQPGIPGMEPDLPA c c c g g c g a g g a g c c g c c c a g c c a g c c c g g c a t t c c g g g c a t g g a a c c a g a c c t g c c a g c c g g g c c g c t c c t c g g c g g g t c g g t c g g g c c g t a a g g c c c g t a c c t t g g t c t g g a c g g t c g g

12,720 180 185 190 195 LTETEEWERRGQQRLPMPDE pBBR1 Rep LTETEEWERRGQQRLPMPDE t t g a c c g a a a c g g a g g a a t g g g a a c g g c g c g g g c a g c a g c g c c t g c c g a t g c c c g a t g a g a a c t g g c t t t g c c t c c t t a c c c t t g c c g c g c c c g t c g t c g c g g a c g g c t a c g g g c t a c t c

PshAI 12,780 200 205 210 215 PCFLDDGEPLEPPTRVTLPR pBBR1 Rep PCFLDDGEPLEPPTRVTLPR c c g t g t t t t c t g g a c g a t g g c g a g c c g t t g g a g c c g c c g a c a c g g g t c a c g c t g c c g c g c g g c a c a a a a g a c c t g c t a c c g c t c g g c a a c c t c g g c g g c t g t g c c c a g t g c g a c g g c g c g

12,840 220 R* pBBR1 Rep R* c g g t a g c a c t t g g g t t g c g c a g c a a c c c g t a a g t g c g c t g t t c c a g a c t a t c g g c t g t a g g c c a t c g t g a a c c c a a c g c g t c g t t g g g c a t t c a c g c g a c a a g g t c t g a t a g c c g a c a t c

3ʹ 5ʹ 12,843 c c g g g c

Restriction Enzymes

Instructions: By default, all cutters are shown. Filter on number of cut sites or search by enzyme name.

Filter

Features

Primers

BLAST

BLAST (Basic Local Alignment Search Tool) finds regions of similarity between biological sequences. Click on the buttons below to submit a BLAST search to NCBI. The results will appear in a new window. See your recent BLAST results on NCBI's website.

  • Nucleotide-Nucleotide BLAST (BLASTN)

  • Translated Nucleotide-Protein BLAST (BLASTX)

  • Sequence alignment using BLAST (BLAST2)

Sequence Analyzer Guide

Map

Displays a graphical map based on nucleotide sequence data labeled with restriction enzymes, plasmid features, ORFs (theoretical open reading frames) and primers. Hovering over data labels will display additional information (e.g. cut site)

To select a portion of sequence, click one location on the plasmid and then a second location to display the sequence between the two locations.

Sequence

Displays both strands of base paired nucleotide sequences with annotated enzymes, plasmid features, ORFs (theoretical open reading frames) and primers. Hovering over data labels will display additional information (e.g. cut site).

To select a portion of sequence, click one location on the sequence and then a second location to display the sequence between the two locations.

Enzymes

List of restriction enzymes that can cut a given nucleotide sequence. Table lists enzyme name and the sequence location of the cut.

Features

List of common features detected in a given nucleotide sequence. Table lists feature name, location, size, color used to indicate its position on the map, and direction (if relevant).

Primers

List of commonly used primers detected in a given nucleotide sequence. Table lists primer name, sequence, length, binding site location, and direction.

BLAST

Use Basic Local Alignment Search Tool (BLAST) via the NCBI website to determine similarity between a given sequence and nucleotide (BLASTN) or protein (BLASTX) sequence databases. Additionally, align a custom nucleotide sequence against a given sequence using BLAST2.

File Downloads

GenBank

File contains the nucleotide sequence and annotated features in GenBank flat file format. Open the file with a text editor or plasmid mapping software to view the sequence.

SnapGene

File contains the nucleotide sequence and enhanced annotations from SnapGene Server. Open the file with SnapGene software or the free Viewer to view the plasmid map, sequence, and perform additional sequence analysis.