Plasmid name: pBI-4 Size: 4358 bp
Resistance: Ampicillin constructed by: Udo Baron, Sabine Freundlieb
Stefan Berger
Host strain: DH5 alpha, E.coli "sure"
Reference: Baron, Udo et al. (1995),Nucleic Acids Res.,17, 3605-3606
The plasmid consists out of four main fragments:
Afl II c/ttaag Esp I gc/tnagc Sna I gta/tac Age I a/ccggt Fse I ggccgg/cc SnaB I tac/gta Avr II c/ctagg Hind III a/agctt Spe I a/ctagt Bcl I t/gatca Nru I tcg/cga Sph I gcatg/c BsaA I yac/gtr Nsi I atgca/t Spl I c/gtacg BspE I t/ccgga Pac I ttaat/taa Sse8337 I cctgca/gg BspM I acctgc 4/8 PaeR7 I c/tcgag Swa I attt/aaat BstB I tt/cgaa Pml I cac/gtg Tth111 I gacn/nngtc Cla I at/cgat Rsr II cg/gwccg Xca I gta/tac Dra III cacnnn/gtg Sap I gcttcttc 1/4 Xcm I ccannnnn/nnnntgg Eco47 III agc/gct Sfi I ggccnnnn/nggcc Xho I c/tcgag EcoN I cctnn/nnnagg SgrA I cr/ccggyg
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 EcoR V gat/atc 632 Nco I c/catgg 774 Sty I c/cwwgg 774 Apa I gggcc/c 1185 Bsp120 I g/ggccc 1185 Bsu36 I cc/tnagg 1288 Bsg I gtgcag 16/14 1294 Msc I tgg/cca 1328 BstX I ccannnnn/ntgg 1331 Bgl II a/gatct 1359 PflM I ccannnn/ntgg 1559 Drd I gacnnnn/nngtc 2085 BceF I acggc 12/13 2469 Bsa I ggtctc 1/5 2943 Fsp I tgc/gca 3096 Pvu I cgat/cg 3243 Xmn I gaann/nnttc 3471 Aat II gacgt/c 3794 PshA I gacnn/nngtc 3871 Nae I gcc/ggc 3912 BstE II g/gtnacc 3955 Bbe I ggcgc/c 3988 Ehe I ggc/gcc 3988 Kas I g/gcgcc 3988 Nar I gg/cgcc 3988 BsaB I gatnn/nnatc 4093 Hpa I gtt/aac 4196 Acc I gt/mkac 4346 Sal I g/tcgac 4346 Not I gc/ggccgc 4352 Eag I c/ggccg 4353
Enzyme Sites Positions AlwN I cagnnn/ctg 2 1228 2394 ApaL I g/tgcac 2 2297 3543 Ase I at/taat 2 1812 3047 Asp718 g/gtacc 2 425 582 BamH I g/gatcc 2 591 4329 BssH II g/cgcgc 2 3931 4034 Fau I cccgc 4/6 2 1832 4014 Kpn I ggtac/c 2 425 582 Mme I tccrac 20/18 2 2173 2357 PpuM I rg/gwccy 2 667 4045 Sac II ccgc/gg 2 17 560 Sca I agt/act 2 936 3354 Sma I ccc/ggg 2 429 586 Tfi I g/awtc 2 1818 1958 Xba I t/ctaga 2 638 4335 Xma I c/ccggg 2 429 586 Afl III a/crygt 3 609 1729 1983 Ava I c/ycgrg 3 429 586 3995 Bbs I gaagac 2/6 3 42 535 3861 Bsm I gaatgc 1/-1 3 3935 4180 4279 Cfr10 I r/ccggy 3 2956 3912 3952 Dsa I c/crygg 3 17 560 774 EcoR I g/aattc 3 7 570 1279 Gsu I ctggag 16/14 3 77 500 2961 Nde I ca/tatg 3 1493 1553 1560 Nsp I rcatg/y 3 1729 1983 3908 Nsp7524 I r/catgy 3 1729 1983 3908 NspC I rcatg/y 3 1729 1983 3908 Ssp I aat/att 3 1038 1072 3678
Aat II 1 BssH II 2 Hga I 9 Pml I - Acc I 1 BstB I - HgiA I 7 PpuM I 2 Afl II - BstE II 1 Hha I 19 PshA I 1 Afl III 3 BstK I 21 Hinc II 4 Pst I 1 Age I - BstN I 10 Hind III - Pvu I 1 Aha II 5 BstU I 16 Hinf I 7 Pvu II 1 Alu I 17 BstX I 1 HinP I 19 Rma I 10 Alw I 15 BstY I 9 Hpa I 1 Rsa I 6 AlwN I 2 Bsu36 I 1 Hpa II 18 Rsr II - Apa I 1 Cfr10 I 3 Hph I 7 Sac I 4 ApaL I 2 Cla I - Kas I 1 Sac II 2 Ase I 2 Csp6 I 6 Kpn I 2 Sal I 1 Asp718 2 Dde I 9 Mae II 5 Sap I - Ava I 3 Dpn I 21 Mae III 13 Sau3A I 21 Ava II 8 Dpn II 21 Mbo I 21 Sau96 I 15 Avr II - Dra I 6 Mbo II 13 Sca I 2 BamH I 2 Dra III - Mcr I 5 ScrF I 21 Ban I 4 Drd I 1 Mlu I 1 SfaN I 7 Ban II 5 Dsa I 3 Mme I 2 Sfe I 5 Bbe I 1 Dsa V 21 Mnl I 37 Sfi I - Bbs I 3 Eae I 5 Msc I 1 SgrA I - Bbv I 12 Eag I 1 Mse I 17 Sma I 2 BceF I 1 Ear I 4 Msp I 18 Sna I - Bcl I - Ecl136 I 4 Nae I 1 SnaB I - Bcn I 11 Eco47 III - Nar I 1 Spe I - Bgl I 4 Eco57 I 5 Nci I 11 Sph I - Bgl II 1 EcoN I - Nco I 1 Spl I - Bsa I 1 EcoO109 I 4 Nde I 3 Sse8337 I - BsaA I - EcoR I 3 Nhe I 1 Ssp I 3 BsaB I 1 EcoR II 10 Nla III 19 Stu I 1 BsaJ I 12 EcoR V 1 Nla IV 22 Sty I 1 Bsg I 1 Ehe I 1 Not I 1 Swa I - BsiE I 5 Esp I - Nru I - Taq I 17 BsiY I 6 Fau I 2 Nsi I - Tfi I 2 Bsm I 3 Fse I - Nsp I 3 Tth111 I - BsmA I 7 Fnu4H I 25 Nsp7524 I 3 Tth111 II 4 Bsp120 I 1 Fok I 8 NspB II 7 Xba I 2 Bsp1286 I 8 Fsp I 1 NspC I 3 Xca I - BspE I - Gdi II 4 Pac I - Xcm I - BspH I 5 Gsu I 3 PaeR7 I - Xho I - BspM I - Hae I 5 PflM I 1 Xma I 2 BspW I 20 Hae II 4 Ple I 5 Xmn I 1 Bsr I 9 Hae III 18
1 80 CTGCAGGAATTCGGGGCCGCGGAGGCTGGATCGGTCCCGGTGTCTTCTATGGAGGTCAAAACAGCGTGGATGGCGTCTCC AGGCGATCTGACGGTTCACTAAACGAGCTCTGCTTATATAGGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTG AAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAA AGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGA GAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAGTCGAGTTTACCACTCCCTATCAGTGA TAGAGAAAAGTGAAAGTCGAGCTCGGTACCCGGGTCGAGTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTT TAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACCTCCATAGAAGACACCGGGACCGATCCAGCCTC CGCGGCCCCGAATTCGAGCTCGGTACCCGGGGATCCTCTAGTCAGCTGACGCGTGCTAGCGCGGCCTCGACGATATCTCT AGACTGAGAACTTCAGGGTGAGTTTGGGGACCCTTGATTGTTCTTTCTTTTTCGCTATTGAAAAATTCATGTTATATGGA GGGGGCAAAGTTTTCAGGGTGTTGTTTAGAATGGGAAGATGTCCCTTGTATCACCATGGACCCTCATGATAATTTTGTTT CTTTCACTTTCTACTCTGTTGACAACCATTGTCTCCTCTTATTTTCTTTTCATTTTCTGTAACTTTTTTCGTTAAACTTT AGCTTGCATTTGTAACGAATTTTTAAATTCACTTTCGTTTATTTGTCAGATTGTAAGTACTTTCTCTAATCACTTTTTTT TCAAGGCAATCAGGGTAATTATATTGTACTTCAGCACAGTTTTAGAGAACAATTGTTATAATTAAATGATAAGGTAGAAT ATTTCTGCATATAAATTCTGGCTGGCGTGGAAATATTCTTATTGGTAGAAACAACTACATCCTGGTAATCATCCTGCCTT TCTCTTTATGGTTACAATGATATACACTGTTTGAGATGAGGATAAAATACTCTGAGTCCAAACCGGGCCCCTCTGCTAAC CATGTTCATGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAACGTGCTGGTTGTTGTGCTGTCTCATCATTTTGGCAAAGA ATTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAG ATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGGTAATAAAGGAAATTT ATTTTCATTGCAATAGTGTGTGGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACA TCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCATATGCTGGCTGCCATGAACAAAGGTGGCTATAAAGAGGT CATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTAT ATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCT CCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGAACTCGACTGCATTAATGAATCGGCCAACGCGCGGGGAGA GGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGT ATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCC AGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAA AATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGT GCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCAAT GCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCC GACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCAC TGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTA GAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAA CAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCC TTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAA GGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGAC AGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGT CGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGG CTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATC CAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTAC AGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGAT CCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCA CTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTC AACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCAC ATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGA TCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAA AACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAAT ATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGG GTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAG GCGTATCACGAGGCCCTTTCGTCTTCACTCGACGCTGGTCGAGCTGATACTTCCCGTCCGCCAGGGGACATGCCGGCGAT GCTGAAGGTCGCGCGCATTCCCGATGAAGAGGCCGGTTACCGCCTGTTGACCTGGTGGGACGGGCAGGGCGCCGCCCGAG TCTTGCCTCGGCGGCGGGCGCTCTGCTCATGGAGCGCGCGTCCGGGGACCTTGCACAGATAGCGTGGTCCGGCCAGACGA CGAGGCTTGCAGGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCC TGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAAT AGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTA TCATGTCTGGATCCTCTAGAACTAGGTCGACGCGGCCG