Plasmid name: pBI-5 Size: 6083 bp
Resistance: Ampicillin constructed by: Udo Baron, Sabine Freundlieb,
Host strain: DH5 alpha, E.coli "sure"
Reference: unpublished
The plasmid consists out of four main fragments:
Afl II c/ttaag Nru I tcg/cga Sna I gta/tac Age I a/ccggt Nsi I atgca/t SnaB I tac/gta Avr II c/ctagg PaeR7 I c/tcgag Sse8337 I cctgca/gg Bcl I t/gatca Pml I cac/gtg Swa I attt/aaat Dra III cacnnn/gtg Rsr II cg/gwccg Tth111 I gacn/nngtc Eco47 III agc/gct Sap I gcttcttc 1/4 Xca I gta/tac Esp I gc/tnagc Sfi I ggccnnnn/nggcc Xho I c/tcgag Fse I ggccgg/cc
Enzyme Position Pst I ctgca/g 1 Stu I agg/cct 457 Pvu II cag/ctg 603 Mlu I a/cgcgt 609 Nhe I g/ctagc 615 Not I gc/ggccgc 621 Eag I c/ggccg 622 Cla I at/cgat 629 Hind III a/agctt 635 Acc I gt/mkac 641 Sal I g/tcgac 641 EcoR V gat/atc 647 Apa I gggcc/c 1200 Bsp120 I g/ggccc 1200 Bsg I gtgcag 16/14 1309 Msc I tgg/cca 1343 BstX I ccannnnn/ntgg 1346 Bgl II a/gatct 1374 PflM I ccannnn/ntgg 1574 Drd I gacnnnn/nngtc 2100 Bsa I ggtctc 1/5 2958 Fsp I tgc/gca 3111 Pvu I cgat/cg 3258 Xmn I gaann/nnttc 3486 Aat II gacgt/c 3809 PshA I gacnn/nngtc 3886 Nae I gcc/ggc 3927 BsaB I gatnn/nnatc 4108 Hpa I gtt/aac 4211 Spe I a/ctagt 4356 EcoN I cctnn/nnnagg 4452 SgrA I cr/ccggyg 4641 BspM I acctgc 4/8 4663 Pac I ttaat/taa 4748 Xcm I ccannnnn/nnnntgg 5333 Sph I gcatg/c 5411 Spl I c/gtacg 5916 BsaA I yac/gtr 5918
Enzyme Sites Positions AlwN I cagnnn/ctg 2 1243 2409 ApaL I g/tgcac 2 2312 3558 Ase I at/taat 2 1827 3062 Asp718 g/gtacc 2 425 582 Bbe I ggcgc/c 2 4003 6038 BceF I acggc 12/13 2 2484 4625 BspE I t/ccgga 2 4859 5375 BssH II g/cgcgc 2 3946 4049 BstB I tt/cgaa 2 5114 5902 BstE II g/gtnacc 2 3970 5462 Bsu36 I cc/tnagg 2 1303 5457 Ehe I ggc/gcc 2 4003 6038 Kas I g/gcgcc 2 4003 6038 Kpn I ggtac/c 2 425 582 Mme I tccrac 20/18 2 2188 2372 Nar I gg/cgcc 2 4003 6038 Nco I c/catgg 2 789 6072 Sac II ccgc/gg 2 17 560 Sca I agt/act 2 951 3369 Sma I ccc/ggg 2 429 586 Sty I c/cwwgg 2 789 6072 Xba I t/ctaga 2 653 4350 Xma I c/ccggg 2 429 586 BamH I g/gatcc 3 591 4344 4362 Bsm I gaatgc 1/-1 3 3950 4195 4294 EcoR I g/aattc 3 7 570 1294 Nde I ca/tatg 3 1508 1568 1575 PpuM I rg/gwccy 3 682 4060 4891 Ssp I aat/att 3 1053 1087 3693
Aat II 1 BssH II 2 Hga I 13 Pml I - Acc I 1 BstB I 2 HgiA I 8 PpuM I 3 Afl II - BstE II 2 Hha I 25 PshA I 1 Afl III 4 BstK I 30 Hinc II 5 Pst I 1 Age I - BstN I 15 Hind III 1 Pvu I 1 Aha II 8 BstU I 21 Hinf I 13 Pvu II 1 Alu I 23 BstX I 1 HinP I 25 Rma I 13 Alw I 22 BstY I 14 Hpa I 1 Rsa I 12 AlwN I 2 Bsu36 I 2 Hpa II 28 Rsr II - Apa I 1 Cfr10 I 5 Hph I 9 Sac I 4 ApaL I 2 Cla I 1 Kas I 2 Sac II 2 Ase I 2 Csp6 I 12 Kpn I 2 Sal I 1 Asp718 2 Dde I 12 Mae II 10 Sap I - Ava I 4 Dpn I 29 Mae III 19 Sau3A I 29 Ava II 9 Dpn II 29 Mbo I 29 Sau96 I 19 Avr II - Dra I 6 Mbo II 24 Sca I 2 BamH I 3 Dra III - Mcr I 6 ScrF I 30 Ban I 5 Drd I 1 Mlu I 1 SfaN I 9 Ban II 6 Dsa I 4 Mme I 2 Sfe I 5 Bbe I 2 Dsa V 30 Mnl I 46 Sfi I - Bbs I 6 Eae I 5 Msc I 1 SgrA I 1 Bbv I 13 Eag I 1 Mse I 23 Sma I 2 BceF I 2 Ear I 7 Msp I 28 Sna I - Bcl I - Ecl136 I 4 Nae I 1 SnaB I - Bcn I 15 Eco47 III - Nar I 2 Spe I 1 Bgl I 4 Eco57 I 6 Nci I 15 Sph I 1 Bgl II 1 EcoN I 1 Nco I 2 Spl I 1 Bsa I 1 EcoO109 I 5 Nde I 3 Sse8337 I - BsaA I 1 EcoR I 3 Nhe I 1 Ssp I 3 BsaB I 1 EcoR II 15 Nla III 22 Stu I 1 BsaJ I 17 EcoR V 1 Nla IV 27 Sty I 2 Bsg I 1 Ehe I 2 Not I 1 Swa I - BsiE I 6 Esp I - Nru I - Taq I 25 BsiY I 11 Fau I 6 Nsi I - Tfi I 5 Bsm I 3 Fse I - Nsp I 4 Tth111 I - BsmA I 9 Fnu4H I 26 Nsp7524 I 4 Tth111 II 6 Bsp120 I 1 Fok I 12 NspB II 8 Xba I 2 Bsp1286 I 10 Fsp I 1 NspC I 4 Xca I - BspE I 2 Gdi II 4 Pac I 1 Xcm I 1 BspH I 5 Gsu I 5 PaeR7 I - Xho I - BspM I 1 Hae I 6 PflM I 1 Xma I 2 BspW I 21 Hae II 5 Ple I 8 Xmn I 1 Bsr I 12 Hae III 21
1 80 CTGCAGGAATTCGGGGCCGCGGAGGCTGGATCGGTCCCGGTGTCTTCTATGGAGGTCAAAACAGCGTGGATGGCGTCTCC AGGCGATCTGACGGTTCACTAAACGAGCTCTGCTTATATAGGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTG AAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAA AGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGA GAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGA TAGAGAAAAGTGAAAGTCGAGCTCGGTACCCGGGTCGAGTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTT TAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACCTCCATAGAAGACACCGGGACCGATCCAGCCTC CGCGGCCCCGAATTCGAGCTCGGTACCCGGGGATCCTCTAGTCAGCTGACGCGTGCTAGCGCGGCCGCATCGATAAGCTT GTCGACGATATCTCTAGACTGAGAACTTCAGGGTGAGTTTGGGGACCCTTGATTGTTCTTTCTTTTTCGCTATTGAAAAA TTCATGTTATATGGAGGGGGCAAAGTTTTCAGGGTGTTGTTTAGAATGGGAAGATGTCCCTTGTATCACCATGGACCCTC ATGATAATTTTGTTTCTTTCACTTTCTACTCTGTTGACAACCATTGTCTCCTCTTATTTTCTTTTCATTTTCTGTAACTT TTTTCGTTAAACTTTAGCTTGCATTTGTAACGAATTTTTAAATTCACTTTCGTTTATTTGTCAGATTGTAAGTACTTTCT CTAATCACTTTTTTTTCAAGGCAATCAGGGTAATTATATTGTACTTCAGCACAGTTTTAGAGAACAATTGTTATAATTAA ATGATAAGGTAGAATATTTCTGCATATAAATTCTGGCTGGCGTGGAAATATTCTTATTGGTAGAAACAACTACATCCTGG TAATCATCCTGCCTTTCTCTTTATGGTTACAATGATATACACTGTTTGAGATGAGGATAAAATACTCTGAGTCCAAACCG GGCCCCTCTGCTAACCATGTTCATGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAACGTGCTGGTTGTTGTGCTGTCTCA TCATTTTGGCAAAGAATTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTC ACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGG TAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTGGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGC AAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCATATGCTGGCTGCCATGAACAAAGG TGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGG TTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCA GATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGAACTCGACTGCATTAATGAATCGGC CAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCG GCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACA TGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCT GACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCC TGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCG TGGCGCTTTCTCAATGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAA CCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCC ACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTA ACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGC TCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGG ATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCA TGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAG TAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGT TGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAG ACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACT TTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGT TGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAA GGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTG GCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGT GACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGG ATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATC TTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGT TTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATAC TCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAA AATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATT AACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCACTCGACGCTGGTCGAGCTGATACTTCCCGTCCGCCAGG GGACATGCCGGCGATGCTGAAGGTCGCGCGCATTCCCGATGAAGAGGCCGGTTACCGCCTGTTGACCTGGTGGGACGGGC AGGGCGCCGCCCGAGTCTTGCCTCGGCGGCGGGCGCTCTGCTCATGGAGCGCGCGTCCGGGGACCTTGCACAGATAGCGT GGTCCGGCCAGACGACGAGGCTTGCAGGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACC TCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGT TACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACT CATCAATGTATCTTATCATGTCTGGATCCTCTAGAACTAGTGGATCCCCCCTAGAGTATTACAATAGCTAAGAATTTCGT CATCGCTGAATACAGTTACATTTTACAATTTGGACTTTCCGCCCTTCTTGGCCTTTATGAGGATCTCTCTGATTTTTCTT GCGTCGAGTTTTCCGGTAAGACCTTTCGGTACTTCGTCCACAAACACAACTCCTCCGCGCAACTTTTTGGCGGTTGTTAC TTGACTGGCGACGTAATCCACGATCTCTTTTTCCGTCATCGTCTTTCCGTGCTCCAAAACAACAACGGCGGCGGGAAGTT CACCGGCGTCATCGTCGGGAAGACCTGCCACGCCCGCGTCGAAGATGTTGGGGTGTTGTAACAATATCGACTCCAATTCA GCGGGGGCCACCTGGTATCCTTTGTATTTAATTAAAGACTTCAAGCGGTCAACTATGAAGAAGTGTTCGTCTTCGTCCCA GTAAGCTATGTCTCCAGAATGTAGCCATCCATCCTTGTCAATCAAGGCGTTGGTCGCTTCCGGATTGTTTACATAACCGG ACATAATCATAGGTCCTCTGACACATAATTCGCCTCTCTGATTAACGCCCAGCGTTTTCCCGGTATCCAGATCCACAACC TTCGCTTCAAAAAATGGAACAACTTTACCGACCGCGCCCGGTTTATCATCCCCCTCGGGTGTAATCAGAATAGCTGATGT AGTCTCAGTGAGCCCATATCCTTGTCGTATCCCTGGAAGATGGAAGCGTTTTGCAACCGCTTCCCCGACTTCTTTCGAAA GAGGTGCGCCCCCAGAAGCAATTTCGTGTAAATTAGATAAATCGTATTTGTCAATCAGAGTGCTTTTGGCGAAGAATGAA AATAGGGTTGGTACTAGCAACGCACTTTGAATTTTGTAATCCTGAAGGGATCGTAAAAACAGCTCTTCTTCAAATCTATA CATTAAGACGACTCGAAATCCACATATCAAATATCCGAGTGTAGTAAACATTCCAAAACCGTGATGGAATGGAACAACAC TTAAAATCGCAGTATCCGGAATGATTTGATTGCCAAAAATAGGATCTCTGGCATGCGAGAATCTGACGCAGGCAGTTCTA TGCGGAAGGGCCACACCCTTAGGTAACCCAGTAGATCCAGAGGAGTTCATTATCAGTGCAATTGTTTTGTCACGATCAAA GGACTCTGGTACAAAATCGTATTCATTAAAACCGGGAGGTAGATGAGATGTGACGAACGTGTACATCGACTGAAATCCCT GGTAATCCGTTTTAGAATCCATGATAATAATTTTCTGGATTATTGGTAATTTTTTTTGCACGTTCAAAATTTTTTGCAAC CCCTTTTTGGAAACAAACACTACGGTAGGCTGCGAAATGTTCATACTGTTGAGCAATTCACGTTCATTATAAATGTCGTT CGCGGGCGCAACTGCAACTCCGATAAATAACGCGCCCAACACCGGCATAAAGAATTGAAGAGAGTTTTCACTGCATACGA CGATTCTGTGATTTGTATTCAGCCCATATCGTTTCATAGCTTCTGCCAACCGAACGGACATTTCGAAGTATTCCGCGTAC GTGATGTTCACCTCGATATGTGCATCTGTAAAAGCAATTGTTCCAGGAACCAGGGCGTATCTCTTCATAGCCTTATGCAG TTGCTCTCCAGCGGTTCCATCCTCTAGCGGATAGAATGGCGCCGGGCCTTTCTTTATGTTTTTGGCGTCTTCCATGGTAC GGG