ID pCal-c-MK standard; circular DNA; ; 8235 BP. DE LMBP 5464. CC Accession number: LMBP 5464 CC (c) BCCM/GeneCorner Plasmid collection (Ghent University, Belgium) CC Last modified: 17-JUL-2012 CC CC VNTAUTHORNAME|BCCM/GeneCorner Plasmid Collection| CC VNTAUTHORTEL|0032-9-33-13-843| CC VNTAUTHOREML|bccm.genecorner@UGent.be| CC VNTAUTHORWWW|www.genecorner.ugent.be| CC VNTAUTHORAD1|Ghent University| CC VNTAUTHORAD2|Technologiepark 927| CC VNTAUTHORAD3|B-9052 Gent-Zwijnaarde Belgium| CC CC VNTREPLTYPE|Plasmid FH Key Location/Qualifiers FH FT promoter 3315..3392 FT /vntifkey="30" FT /label=lacI\promoter FT /note="E. coli repressor lacI promoter, incl. ribosome binding site" FT RBS complement(2921..2963) FT /vntifkey="32" FT /label=5'\UTR\of\T7\gene\10,\incl.\RBS FT promoter complement(2965..3006) FT /vntifkey="30" FT /label=T7\promoter\with\lac\operator FT CDS 3393..4475 FT /vntifkey="4" FT /label=lacI FT CDS 5787..5978 FT /vntifkey="4" FT /label=rop FT /note="modulation repressor coding sequence" FT rep_origin complement(6224..6626) FT /vntifkey="33" FT /label=pMB1\ori FT CDS complement(7167..8027) FT /vntifkey="4" FT /label=ampicillin\resistance FT terminator complement(213..260) FT /vntifkey="43" FT /label=T7\terminator FT misc_marker complement(325..402) FT /vntifkey="22" FT /label=calmodulin-binding\peptide\tag FT misc_marker complement(409..426) FT /vntifkey="22" FT /label=thrombin\recognition\site FT CDS complement(433..2904) FT /vntifkey="4" FT /label=hMLT FT mutation complement(955..957) FT /vntifkey="62" FT /label=PCR\artefact FT /note="silent mutation; ACA was replaced by ACG" FT mutation complement(1417..1419) FT /vntifkey="62" FT /label=stability\creator FT /note="silent mutation; wt: GCA ('GCA'GAAG recombines with the same heptamer in hAPI2)" FT mutation complement(2899..2901) FT /vntifkey="62" FT /label=Kozak FT /note="TCG was replaced by GCG (S2A) creating Kozak consensus sequence GCCACCATGG" SQ Sequence 8235 BP; 1982 A; 2152 C; 2025 G; 2076 t; ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa 60 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg 120 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt 180 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag 240 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt 300 cgggctttgt tagcagccgg atcaaagtgc cccggaggat gagattttct taaagcggtt 360 ggctgctgag acggctatga aattcttttt ccatcgtcgc ttggtaccat tcccacgtgg 420 atacatggat cctttttcag aaattctgag cctgtcagag aaactaaatg gtatttcatc 480 agttgtctct actggcacat tacttctact aaaatgacat gaagcatggt gagcgaaact 540 tgaaataaat gcatctggag tccggctgca atgacagcta tctgctggtg taacattact 600 tggattacca ggatgtgaat ggtaaacacc atggaatggg tcttgcaatg agtgataatg 660 ccctgctcct cctgaggtgg ctgcagaact ctggtaagga ccattagaca taagacaagt 720 ttgaaagcaa gtcttccttc ccaaacctcg atgcatgtct aatttagcaa tgagaggttt 780 cccaacattc acttcctgct tgtcctctac agtatcttcc aatcctgagt actgatatga 840 taaacatact gtgaagacta gatgttcctt taatttttgc agtgaactga gtctggtata 900 gaggcaatgc ttgggaagat cctttgatac caagtagctg ccagtttctt caggcgtgcc 960 tttatttgca tcttttggat caatatctag atcaagtgga aaatcagtaa cgtaggcatc 1020 acacattatt atctccggtg gtttgtaaac tatacttgta tagatgatca tgacattgga 1080 aaactcagct gcaaatccta attgaatctg aacaccacag tcaaacttaa gacacatact 1140 ttctggaagt tcatgagcct tggcccactg tagattccgc acaagagatt cagcagaata 1200 ttctgttccc tgtattggat cagtaagtgc tctcttctca gataaactac ttcgaatctc 1260 tagagcctgt ttgcctttgg taaggtgaca cttacccata tcttctgcaa cttcatccag 1320 taacacagtg attttcttat cttctaataa tctgtctttt aaaaatttca taaagattcc 1380 atttgccaat ccagaatgct ggatttcaaa agcttcagct ccttgacacg tggcatatcc 1440 aaacacaata ttggcggtga cttttagtgc atccaagatt ggaatggtat catcgtagtc 1500 atttcttttc ctacacatat ccaataagaa cacattaagt ccagtttctt tttcttgcat 1560 caatttcagt atattttgta cacacagaca attttcagac ctatatggat ttggagcatc 1620 aacggggacc atgaagctgt tcccaaaatt ttcataacca tgtcctgcat aatataataa 1680 cccatatact cccttgtcta aaaggagtaa aaactcatcc acagcattac gcatctcata 1740 ttcagtaaga tccaacagtg aaaccacttt gaagtccagc tgtctcagta agttagtcaa 1800 ttcgtacaca tccaccaaag gagctttgag cttggggtgc tcccggtaat tcatatttcc 1860 tatcaaaagg gcaaccttgt ccttcgccaa aggctggtca gttgtttgct ctttattatc 1920 aggatgacca agattattta attcatcttc agtgcactcc actgcctcat ctgttcttcc 1980 tatgatgatt tctaccttct tgctatcttg actgtctcga tcattatata catgacacca 2040 gtaggttcct tggtgttcca aatccacata aggcaccatg tatagctttt tggtctcatg 2100 tgttaatggt aattcatttt tgaaccactg gtagtgagga atagggcttc caacagcaac 2160 acactgtaaa accaatgtgc tgcctggcat cagcttttgg gaagttggtt caacacagat 2220 ttgcaacttg gattcagaga cgccatcaac acttctctgg aagctctctg ggatgtcgca 2280 aacatccagc tgtgaccact ggctgaattc aaaggtgaaa ttgttattaa ctcgacagac 2340 ataaaagcct gcatctttta catgcactgc attaaaaata agctctgatg tatttccatt 2400 tggaatctct ttattcattt tgaaccactg atattgaaca aaaggatgtc cagttgcccg 2460 gcaacacagt ttcacaaact gtccagccaa gactgccttt gactctgggt ttacagtaat 2520 ctttattcct ggggggctga gaagctgaag aacttcagtg tgttccatag cctgcaggaa 2580 atcactcaat tctgtgactg tgcaaccttt ttcacccatt aacttcagca gacacaggct 2640 ggggcttcct tcaggctcca gtaccttaag agaacactgc tccaggtcta ggcaactgag 2700 gcggaggcgc ccgcgactcc ccgccagctc cgccagtctc ctccagcccc ggccctcggg 2760 cgcctgatcc aggagctcgc tgagcctccg cagcagcggc tcccgcaggc ggttgagggt 2820 cgcgccggcc ggaggggcga gcagcggccc cgtgggggcg gccgagggcg gcagggcctg 2880 tagcgggtcc cccaacagcg ccatggtggc aggatccatg gtatatctcc ttcttaaagt 2940 taaacaaaat tatttctaga ggggaattgt tatccgctca caattcccct atagtgagtc 3000 gtattaattt cgcgggatcg agatctcgat cctctacgcc ggacgcatcg tggccggcat 3060 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga 3120 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc 3180 cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt 3240 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga 3300 gcgtcgagat cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc 3360 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg 3420 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg 3480 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca 3540 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca 3600 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac 3660 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg 3720 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg 3780 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg 3840 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg 3900 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa 3960 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa 4020 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca 4080 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg 4140 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg 4200 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca 4260 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg 4320 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc 4380 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 4440 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag 4500 ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc 4560 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc 4620 atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt 4680 cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc 4740 gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc 4800 gccggcatgg cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg 4860 atggccttcc ccattatgat tcttctcgct tccggcggca tcgggatgcc cgcgttgcag 4920 gccatgctgt ccaggcaggt agatgacgac catcagggac agcttcaagg atcgctcgcg 4980 gctcttacca gcctaacttc gatcactgga ccgctgatcg tcacggcgat ttatgccgcc 5040 tcggcgagca catggaacgg gttggcatgg attgtaggcg ccgccctata ccttgtctgc 5100 ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct cgacctgaat ggaagccggc 5160 ggcacctcgc taacggattc accactccaa gaattggagc caatcaattc ttgcggagaa 5220 ctgtgaatgc gcaaaccaac ccttggcaga acatatccat cgcgtccgcc atctccagca 5280 gccgcacgcg gcgcatctcg ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc 5340 tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat 5400 caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa 5460 caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc 5520 cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac 5580 ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc 5640 gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat 5700 cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca 5760 gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg 5820 cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg 5880 atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct 5940 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg 6000 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg 6060 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata 6120 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tatgcggtgt 6180 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg 6240 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 6300 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 6360 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 6420 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 6480 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 6540 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 6600 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 6660 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 6720 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 6780 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 6840 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 6900 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 6960 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 7020 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 7080 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 7140 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 7200 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 7260 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 7320 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 7380 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 7440 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca 7500 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 7560 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 7620 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 7680 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 7740 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg 7800 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 7860 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 7920 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 7980 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 8040 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 8100 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 8160 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc 8220 tttcgtcttc aagaa 8235 //