Plasmid name: pUHD 15-20 Size: 3872 bp Resistance: Ampicillin constructed by: Manfred Gossen Host strain: XL1 blue, DH5 alpha, E.coli "SURE" Reference: Manfred Gossen unpublished
The plasmid consists of three main fragments:
Afl II c/ttaag Dra III cacnnn/gtg Pml I cac/gtg
Age I a/ccggt Eag I c/ggccg Sap I gcttcttc 1/4
Apa I gggcc/c EcoR V gat/atc Sfi I ggccnnnn/nggcc
Asp718 g/gtacc Esp I gc/tnagc SgrA I cr/ccggyg
Avr II c/ctagg Fse I ggccgg/cc Sna I gta/tac
Bgl II a/gatct Kpn I ggtac/c Spe I a/ctagt
Bsp120 I g/ggccc Msc I tgg/cca Sse8337 I cctgca/gg
BspE I t/ccgga Nco I c/catgg Stu I agg/cct
BspM I acctgc 4/8 Nhe I g/ctagc Swa I attt/aaat
BstX I ccannnnn/ntgg Not I gc/ggccgc Tth111 I gacn/nngtc
Bsu36 I cc/tnagg Pac I ttaat/taa Xca I gta/tac
Cla I at/cgat PflM I ccannnn/ntgg Xcm I ccannnnn/nnnntgg
Enzyme Position PaeR7 I c/tcgag 1 Xho I c/tcgag 1 BstB I tt/cgaa 41 Rsr II cg/gwccg 71 Mlu I a/cgcgt 102 Pst I ctgca/g 131 EcoN I cctnn/nnnagg 391 BsaA I yac/gtr 422 SnaB I tac/gta 422 Nru I tcg/cga 465 Nsi I atgca/t 572 Eco47 III agc/gct 589 Bsg I gtgcag 16/14 728 Spl I c/gtacg 823 Dsa I c/crygg 929 Sac II ccgc/gg 929 Sal I g/tcgac 951 PshA I gacnn/nngtc 966 Ban II grgcy/c 988 Ecl136 I gag/ctc 988 Sac I gagct/c 988 Sph I gcatg/c 1023 Sma I ccc/ggg 1077 Xma I c/ccggg 1077 BamH I g/gatcc 1204 Hpa I gtt/aac 1337 PpuM I rg/gwccy 1487 Bbe I ggcgc/c 1545 Ehe I ggc/gcc 1545 Kas I g/gcgcc 1545 Nar I gg/cgcc 1545 BstE II g/gtnacc 1577 Nae I gcc/ggc 1621 Hind III a/agctt 1662 Pvu II cag/ctg 1810 Drd I gacnnnn/nngtc 2090 AlwN I cagnnn/ctg 2399 Bsa I ggtctc 1/5 2948 Gsu I ctggag 16/14 2966 Fsp I tgc/gca 3101 Pvu I cgat/cg 3248 Sca I agt/act 3359 Xmn I gaann/nnttc 3476 Ssp I aat/att 3683 Aat II gacgt/c 3799 Bbs I gaagac 2/6 3866
Enzyme Sites Positions: Acc I gt/mkac 2 843 951 ApaL I g/tgcac 2 2302 3548 Ase I at/taat 2 1817 3052 Bcl I t/gatca 2 717 761 Bgl I gccnnnn/nggc 2 1523 2995 BsaB I gatnn/nnatc 2 1042 1436 BsmA I gtctc 1/5 2 2948 3713 EcoR I g/aattc 2 38 179 Nde I ca/tatg 2 184 765 Sty I c/cwwgg 2 723 1167 Xba I t/ctaga 2 175 190 Afl III a/crygt 3 102 1055 1988 Ava I c/ycgrg 3 1 1077 1538 Ban I g/gyrcc 3 371 1545 2829 BspH I t/catga 3 2708 3716 3821 BssH II g/cgcgc 3 818 1499 1602 Cfr10 I r/ccggy 3 1581 1621 2961 Eco57 I ctgaag 16/14 3 1611 2515 3563 EcoO109 I rg/gnccy 3 854 1487 3856 Fok I ggatg 9/13 3 2861 3042 3329 Mme I tccrac 20/18 3 244 2178 2362 Tth111 II caarca 11/9 3 2562 2595 2601
Aat II 1 BssH II 3 Hga I 10 Pml I - Acc I 2 BstB I 1 HgiA I 4 PpuM I 1 Afl II - BstE II 1 Hha I 29 PshA I 1 Afl III 3 BstK I 14 Hinc II 4 Pst I 1 Age I - BstN I 6 Hind III 1 Pvu I 1 Aha II 5 BstU I 23 Hinf I 9 Pvu II 1 Alu I 18 BstX I - HinP I 29 Rma I 9 Alw I 13 BstY I 9 Hpa I 1 Rsa I 7 AlwN I 1 Bsu36 I - Hpa II 17 Rsr II 1 Apa I - Cfr10 I 3 Hph I 7 Sac I 1 ApaL I 2 Cla I - Kas I 1 Sac II 1 Ase I 2 Csp6 I 7 Kpn I - Sal I 1 Asp718 - Dde I 10 Mae II 6 Sap I - Ava I 3 Dpn I 22 Mae III 11 Sau3A I 22 Ava II 8 Dpn II 22 Mbo I 22 Sau96 I 14 Avr II - Dra I 4 Mbo II 11 Sca I 1 BamH I 1 Dra III - Mcr I 5 ScrF I 14 Ban I 3 Drd I 1 Mlu I 1 SfaN I 10 Ban II 1 Dsa I 1 Mme I 3 Sfe I 5 Bbe I 1 Dsa V 14 Mnl I 25 Sfi I - Bbs I 1 Eae I 4 Msc I - SgrA I - Bbv I 13 Eag I - Mse I 18 Sma I 1 BceF I 5 Ear I 4 Msp I 17 Sna I - Bcl I 2 Ecl136 I 1 Nae I 1 SnaB I 1 Bcn I 8 Eco47 III 1 Nar I 1 Spe I - Bgl I 2 Eco57 I 3 Nci I 8 Sph I 1 Bgl II - EcoN I 1 Nco I - Spl I 1 Bsa I 1 EcoO109 I 3 Nde I 2 Sse8337 I - BsaA I 1 EcoR I 2 Nhe I - Ssp I 1 BsaB I 2 EcoR II 6 Nla III 14 Stu I - BsaJ I 11 EcoR V - Nla IV 16 Sty I 2 Bsg I 1 Ehe I 1 Not I - Swa I - BsiE I 5 Esp I - Nru I 1 Taq I 15 BsiY I 9 Fau I 8 Nsi I 1 Tfi I 4 Bsm I 4 Fse I - Nsp I 5 Tth111 I - BsmA I 2 Fnu4H I 26 Nsp7524 I 5 Tth111 II 3 Bsp120 I - Fok I 3 NspB II 5 Xba I 2 Bsp1286 I 4 Fsp I 1 NspC I 5 Xca I - BspE I - Gdi II 4 Pac I - Xcm I - BspH I 3 Gsu I 1 PaeR7 I 1 Xho I 1 BspM I - Hae I 4 PflM I - Xma I 1 BspW I 18 Hae II 6 Ple I 5 Xmn I 1 Bsr I 9 Hae III 18
1 80 CTCGAGATCCGGCCCCGCCCAGCGTCTTGTCATTGGCGAATTCGAACACGCAGATGCAGTCGGGGCGGCGCGGTCCGAGG TCCACTTCGCATATTAAGGTGACGCGTGTGGCCTCGAACACCGAGCGACCCTGCAGCGACCCGCTTAACAGCGTCAACAG CGTGCCGCAGATCCTCTAGAATTCATATGTCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAAT GAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGT AAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAG GGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAA GTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTT TTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATC AAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGAT CACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAG TGGGTCCGCGTACAGCCGCGCGCGTACGAAAAACAATTACGGGTCTACCATCGAGGGCCTGCTCGATCTCCCGGACGACG ACGCCCCCGAAGAGGCGGGGCTGGCGGCTCCGCGCCTGTCCTTTCTCCCCGCGGGACACACGCGCAGACTGTCGACGGCC CCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGA CGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCG CTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGTAGGGGGCG CGAGGATCCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTT GTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCAT TTTATGTTTCAGGTTCAGGGGGAGGTGTGGGAGGTTTTTTAAAGCAAGTAAAACCTCTACAAATGTGGTATGGCTGATTA TGATCCTGCAAGCCTCGTCGTCTGGCCGGACCACGCTATCTGTGCAAGGTCCCCGGACGCGCGCTCCATGAGCAGAGCGC CCGCCGCCGAGGCAAGACTCGGGCGGCGCCCTGCCCGTCCCACCAGGTCAACAGGCGGTAACCGGCCTCTTCATCGGGAA TGCGCGCGACCTTCAGCATCGCCGGCATGTCCCCTGGCGGACGGGAAGTATCAGCTCGACCAAGCTTGGCGAGATTTTCA GGAGCTAAGGAAGCTAAAATGGAGAAAAAAATCACTGGATATACCACCGTTGATATATCCCAATGGCATCGTAAAGAACA TTTTGAGGCATTTCAGTCAGTTGCTCAATGTACCTATAACCAGACCGTTCAGCTGCATTAATGAATCGGCCAACGCGCGG GGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGA GCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAA AGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATC ACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCC CTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTC TCAATGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTC AGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCA GCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTA CACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCG GCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAA GATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATC AAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGT CTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTC CCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTC ACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCT CCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATT GCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTAC ATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGT TATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAG TACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGC GCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGT TGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGA GCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTT TCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAA TAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAA AATAGGCGTATCACGAGGCCCTTTCGTCTTCA