ID pCal-c-A7M8K standard; circular DNA; ; 8601 BP. DE LMBP 5462. CC Accession number: LMBP 5462 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 RBS complement(3287..3329) FT /vntifkey="32" FT /label=5'\UTR\of\T7\gene\10,\incl.\RBS FT promoter complement(3331..3372) FT /vntifkey="30" FT /label=T7\promoter\with\lac\operator FT promoter 3681..3758 FT /vntifkey="30" FT /label=lacI\promoter FT /note="E. coli repressor lacI promoter, incl. ribosome binding site" FT CDS complement(433..1946) FT /vntifkey="4" FT /label=hMLT\fragment FT /note="exon 8 --> end" FT CDS complement(1947..3270) FT /vntifkey="4" FT /label=hAPI2\fragment FT /note="exon 1 --> 7" FT CDS 3759..4841 FT /vntifkey="4" FT /label=lacI FT CDS 6153..6344 FT /vntifkey="4" FT /label=rop FT /note="modulation repressor coding sequence" FT rep_origin complement(6590..6992) FT /vntifkey="33" FT /label=pMB1\ori FT CDS complement(7533..8393) 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 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(2017..2019) FT /vntifkey="62" FT /label=PCR\artefact FT /note="silent mutation; TTA was replaced by TTG" FT mutation complement(3265..3267) FT /vntifkey="62" FT /label=Kozak FT /note="AAC was replaced by GAC (N2D) creating Kozak consensus sequence GCCACCATGG" SQ Sequence 8601 BP; 2123 A; 2178 C; 2062 G; 2238 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 attcatttga ttctttttcc tcagttgctc tttctctctc 1980 ctcttccctt atttcatctt ctgcattgag taagtccaac acaagatcat tgactagtct 2040 ataattctct ccagttgcta ggatttttct ctgaactgtc tgttttacca ggcttctact 2100 aaagcccatt tccacggcag cattaatcac aggagtattc atcatgattg catcttctga 2160 atggtcttct ccaggttcaa aatggataat tgatgactct gcattttcat ctcctgggct 2220 gtctgatgtg gatagcagct gttcaagtag atgagggtaa ctggcttgaa cttgacggat 2280 gaactcctgt cctttaattc ttatcaagta ctcacacctt ggaaaccact tggcatgttg 2340 aacccatgga tcatctccag attcccaaca cctgagtcca ccatcacagc aaaagcattt 2400 gacatcatca ctgttaccca cataataaaa acccgcactt gcaagctgct caggattaac 2460 tagaacacta gagggccagt taaagaatgt tttaaagcgg gctgcatgtg tctgcatgct 2520 cagattagaa actgtgtatc ttgaagtgtc ttgaagctga ttttctataa atgggcattt 2580 gggaaaatgt ctcaggtgtt ctgacatagc attatccttc ggttcccaat tgctcaattt 2640 tccaccacag gcaaagcaag ccactctgtc tccaggtcct atgtagtaaa agcctgcttt 2700 tgccagatct gttggcgaca gaaaagtcaa tggccatgtc tgaaaagtaa gtaatctggc 2760 attttcgtta ttcattgcac agtggtagga acttctcatc aaggcagaaa aatcttgatt 2820 tgctctggag tttacaggat ttgatggaga gtttgaataa gagccacgga aatatccact 2880 gttttctgta cccggaagta atgagtgtgt ggaatttgtt actgaagaag gaaaagtagg 2940 ctgagaggta gcttccaagt tgttaacgga atttagactc tgaacgaatc tgcagctagg 3000 atacaacttt ttatgctttt cagtaggact gtctcctctt ttccagttat ccagcatcag 3060 gccacaacag aagcatttga ccttgtcatt cacaccagtg taatagaaac cagcacgagc 3120 aagactcctt tctgagacag gaaccccagc aggaaaagtg gaatacgtag acattcggta 3180 cagttcacat gacaagtcgt atttcagttc aaacgtgttg gcgcttttca tcaaatttga 3240 taagaatatg ctgttttcta ctatgtccat ggtggcagga tccatggtat atctccttct 3300 taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag 3360 tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc 3420 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg 3480 ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc 3540 aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc 3600 ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa 3660 gggagagcgt cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat 3720 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg 3780 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca 3840 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca 3900 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca 3960 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg 4020 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta 4080 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc 4140 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc 4200 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc 4260 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc 4320 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca 4380 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac 4440 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc 4500 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata 4560 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca 4620 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct 4680 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa 4740 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc 4800 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta 4860 agttagctca ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca 4920 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc 4980 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac 5040 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac 5100 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc 5160 attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga 5220 ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg 5280 ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg 5340 ctcgcggctc ttaccagcct aacttcgatc actggaccgc tgatcgtcac ggcgatttat 5400 gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc cctatacctt 5460 gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctgaatggaa 5520 gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat caattcttgc 5580 ggagaactgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg tccgccatct 5640 ccagcagccg cacgcggcgc atctcgggca gcgttgggtc ctggccacgg gtgcgcatga 5700 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga 5760 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct 5820 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt 5880 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg 5940 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt 6000 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt 6060 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca 6120 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta 6180 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg 6240 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc 6300 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg 6360 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt 6420 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag 6480 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatatg 6540 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct 6600 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 6660 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 6720 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 6780 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 6840 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 6900 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 6960 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 7020 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 7080 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 7140 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 7200 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 7260 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 7320 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 7380 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 7440 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 7500 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 7560 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 7620 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 7680 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 7740 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 7800 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg 7860 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 7920 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 7980 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 8040 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 8100 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat 8160 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 8220 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 8280 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 8340 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 8400 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 8460 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 8520 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg 8580 aggccctttc gtcttcaaga a 8601 //