Plasmid name: pUHG17-1 Size: 5021 bp Resistance: Ampicillin constructed by: Manfred Gossen Host strain: XL-1 Blue, DH5 alpha, E.coli "SURE" Reference: Gossen, M. et al (1995),Science, 268, 1766-1769
The plasmid consists of three main fragments:
Afl II c/ttaag Ehe I ggc/gcc Pst I ctgca/g
Age I a/ccggt Esp I gc/tnagc Pvu II cag/ctg
Asp718 g/gtacc Fse I ggccgg/cc Rsr II cg/gwccg
Avr II c/ctagg Hpa I gtt/aac Sap I gcttcttc 1/4
Bbe I ggcgc/c Kas I g/gcgcc Sfi I ggccnnnn/nggcc
Bsm I gaatgc 1/-1 Kpn I ggtac/c SgrA I cr/ccggyg
BspE I t/ccgga Nae I gcc/ggc Sna I gta/tac
BspM I acctgc 4/8 Nar I gg/cgcc Sse8337 I cctgca/gg
BstB I tt/cgaa Nhe I g/ctagc Stu I agg/cct
BstE II g/gtnacc Not I gc/ggccgc Swa I attt/aaat
Cla I at/cgat Nru I tcg/cga Tth111 I gacn/nngtc
Dra III cacnnn/gtg Nsi I atgca/t Xca I gta/tac
Eag I c/ggccg Pac I ttaat/taa Xcm I ccannnnn/nnnntgg
EcoR V gat/atc Pml I cac/gtg
Enzyme Position PaeR7 I c/tcgag 1 Xho I c/tcgag 1 Spe I a/ctagt 103 BsaA I yac/gtr 442 SnaB I tac/gta 442 Bbs I gaagac 2/6 731 Hind III a/agctt 869 EcoN I cctnn/nnnagg 977 Mlu I a/cgcgt 1158 Eco47 III agc/gct 1175 BssH II g/cgcgc 1404 Spl I c/gtacg 1409 Sal I g/tcgac 1537 PshA I gacnn/nngtc 1552 Sph I gcatg/c 1609 BsaB I gatnn/nnatc 1628 Sma I ccc/ggg 1663 Xma I c/ccggg 1663 BamH I g/gatcc 1790 PpuM I rg/gwccy 1825 Apa I gggcc/c 2343 Bsp120 I g/ggccc 2343 Bsu36 I cc/tnagg 2446 Msc I tgg/cca 2486 Bgl II a/gatct 2517 PflM I ccannnn/ntgg 2717 Drd I gacnnnn/nngtc 3243 Bsa I ggtctc 1/5 4101 Cfr10 I r/ccggy 4114 Fsp I tgc/gca 4254 Pvu I cgat/cg 4401 Xmn I gaann/nnttc 4629
Enzyme Sites Positions: Acc I gt/mkac 2 1429 1537 AlwN I cagnnn/ctg 2 2386 3552 ApaL I g/tgcac 2 3455 4701 Ava I c/ycgrg 2 1 1663 Bcl I t/gatca 2 1303 1347 Bsg I gtgcag 16/14 2 1314 2452 BstX I ccannnnn/ntgg 2 865 2489 Ecl136 I gag/ctc 2 668 1574 EcoR I g/aattc 2 765 2437 Gsu I ctggag 16/14 2 696 4119 Nco I c/catgg 2 464 1932 Sac I gagct/c 2 668 1574 Sac II ccgc/gg 2 756 1515 Sca I agt/act 2 2094 4512 Xba I t/ctaga 2 776 1796 Ase I at/taat 3 110 2970 4205 Ban I g/gyrcc 3 569 957 3982 Ban II grgcy/c 3 668 1574 2343 Gdi II yggccg -5/-1 3 1716 2980 4422 Hinc II gty/rac 3 86 1537 1976 Ssp I aat/att 3 2196 2230 4836
Aat II 5 BssH II 1 Hga I 8 Pml I - Acc I 2 BstB I - HgiA I 5 PpuM I 1 Afl II - BstE II - Hha I 22 PshA I 1 Afl III 4 BstK I 19 Hinc II 3 Pst I - Age I - BstN I 10 Hind III 1 Pvu I 1 Aha II 9 BstU I 19 Hinf I 10 Pvu II - Alu I 17 BstX I 2 HinP I 22 Rma I 9 Alw I 11 BstY I 8 Hpa I - Rsa I 14 AlwN I 2 Bsu36 I 1 Hpa II 14 Rsr II - Apa I 1 Cfr10 I 1 Hph I 9 Sac I 2 ApaL I 2 Cla I - Kas I - Sac II 2 Ase I 3 Csp6 I 14 Kpn I - Sal I 1 Asp718 - Dde I 13 Mae II 13 Sap I - Ava I 2 Dpn I 22 Mae III 14 Sau3A I 22 Ava II 7 Dpn II 22 Mbo I 22 Sau96 I 18 Avr II - Dra I 6 Mbo II 13 Sca I 2 BamH I 1 Dra III - Mcr I 5 ScrF I 19 Ban I 3 Drd I 1 Mlu I 1 SfaN I 8 Ban II 3 Dsa I 4 Mme I 4 Sfe I 5 Bbe I - Dsa V 19 Mnl I 34 Sfi I - Bbs I 1 Eae I 4 Msc I 1 SgrA I - Bbv I 12 Eag I - Mse I 21 Sma I 1 BceF I 5 Ear I 4 Msp I 14 Sna I - Bcl I 2 Ecl136 I 2 Nae I - SnaB I 1 Bcn I 9 Eco47 III 1 Nar I - Spe I 1 Bgl I 5 Eco57 I 4 Nci I 9 Sph I 1 Bgl II 1 EcoN I 1 Nco I 2 Spl I 1 Bsa I 1 EcoO109 I 4 Nde I 6 Sse8337 I - BsaA I 1 EcoR I 2 Nhe I - Ssp I 3 BsaB I 1 EcoR II 10 Nla III 23 Stu I - BsaJ I 12 EcoR V - Nla IV 20 Sty I 4 Bsg I 2 Ehe I - Not I - Swa I - BsiE I 5 Esp I - Nru I - Taq I 13 BsiY I 11 Fau I 8 Nsi I - Tfi I 4 Bsm I - Fse I - Nsp I 5 Tth111 I - BsmA I 7 Fnu4H I 22 Nsp7524 I 5 Tth111 II 4 Bsp120 I 1 Fok I 7 NspB II 5 Xba I 2 Bsp1286 I 6 Fsp I 1 NspC I 5 Xca I - BspE I - Gdi II 3 Pac I - Xcm I - BspH I 5 Gsu I 2 PaeR7 I 1 Xho I 1 BspM I - Hae I 4 PflM I 1 Xma I 1 BspW I 21 Hae II 4 Ple I 6 Xmn I 1 Bsr I 10 Hae III 20
1 80 CTCGAGGAGCTTGGCCCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACATTACC GCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGT TCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACG TATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGCTAAACTGCCCACTTGGC AGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCC AGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTT TGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAG TTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGC GTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTT GACCTCCATAGAAGACACCGGGACCGATCCAGCCTCCGCGGCCCGAATTCATATGTCTAGATTAGATAAAAGTAAAGTGA TTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTTGGTGTA GAGCAGCCTACACTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCA CCATACTCACTTTTGCCCTTTAAAAGGGGAAAGCTGGCAAGATTTTTTACGCAATAACGCTAAAAGTTTTAGATGTGCTT TACTAAGTCATCGCAATGGAGCAAAAGTACATTCAGATACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAA TTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAACGCGTTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGG TTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTAT TACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGA TTAGAAAAACAACTTAAATGTGAAAGTGGGTCCGCGTACAGCCGCGCGCGTACGAAAAACAATTACGGGTCTACCATCGA GGGCCTGCTCGATCTCCCGGACGACGACGCCCCCGAAGAGGCGGGGCTGGCGGCTCCGCGCCTGTCCTTTCTCCCCGCGG GACACACGCGCAGACTGTCGACGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTG GCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTAC CCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAA TTGACGAGTACGGTGGGTAGGGGGCGCGAGGATCCTCTAGACTGAGAACTTCAGGGTGAGTTTGGGGACCCTTGATTGTT CTTTCTTTTTCGCTATTGAAAAATTCATGTTATATGGAGGGGGCAAAGTTTTCAGGGTGTTGTTTAGAATGGGAAGATGT CCCTTGTATCACCATGGACCCTCATGATAATTTTGTTTCTTTCACTTTCTACTCTGTTGACAACCATTGTCTCCTCTTAT TTTCTTTTCATTTTCTGTAACTTTTTTCGTTAAACTTTAGCTTGCATTTGTAACGAATTTTTAAATTCACTTTCGTTTAT TTGTCAGATTGTAAGTACTTTCTCTAATCACTTTTTTTTCAAGGCAATCAGGGTAATTATATTGTACTTCAGCACAGTTT TAGAGAACAATTGTTATAATTAAATGATAAGGTAGAATATTTCTGCATATAAATTCTGGCTGGCGTGGAAATATTCTTAT TGGTAGAAACAACTACATCCTGGTAATCATCCTGCCTTTCTCTTTATGGTTACAATGATATACACTGTTTGAGATGAGGA TAAAATACTCTGAGTCCAAACCGGGCCCCTCTGCTAACCATGTTCATGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAAC GTGCTGGTTGTTGTGCTGTCTCATCATTTTGGCAAAGAATTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGC TGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGC CCCTTGAGCATCTGACTTCTGGGTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTGGGAATTTTTTGTGTCTCTCA CTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCAT ATGCTGGCTGCCATGAACAAAGGTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTC CATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATT TTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGAA CTCGACTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACT GACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATC AGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGT TTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTA TAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTC CGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCAATGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCT CCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAAC CCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTAC AGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTA CCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAG CAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAA CTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTA AATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATC TGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCC CAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCG AGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCG CCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATT CAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTC CGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATG CCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTG CTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTT CGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCA GCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGAC ACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGAT ACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAA GAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTC