ID pCal-n-MK standard; circular DNA; ; 8251 BP. DE LMBP 5466. CC Accession number: LMBP 5466 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 3331..3408 FT /vntifkey="30" FT /label=lacI\promoter FT /note="E. coli repressor lacI promoter, incl. ribosome binding site" FT promoter complement(2981..3022) FT /vntifkey="30" FT /label=T7\promoter\with\lac\operator FT RBS complement(2935..2979) FT /vntifkey="32" FT /label=5'\UTR\of\T7\gene\10,\incl.\RBS FT CDS 3409..4491 FT /vntifkey="4" FT /label=lacI FT CDS 5803..5994 FT /vntifkey="4" FT /label=rop FT /note="modulation repressor coding sequence" FT rep_origin complement(6240..6642) FT /vntifkey="33" FT /label=pMB1\ori FT CDS complement(7183..8043) FT /vntifkey="4" FT /label=ampicillin\resistance FT terminator complement(213..260) FT /vntifkey="43" FT /label=T7\terminator FT misc_marker complement(2836..2853) FT /vntifkey="22" FT /label=thrombin\recognition\site FT misc_marker complement(2854..2931) FT /vntifkey="22" FT /label=calmodulin-binding\peptide\tag FT mutation complement(853..855) FT /vntifkey="62" FT /label=PCR\artefact FT /note="silent mutation; ACA was replaced by ACG" FT mutation complement(1315..1317) FT /vntifkey="62" FT /label=stability\creator FT /note="silent mutation; wt: GCA ('GCA'GAAG recombines with the same heptamer in hAPI2)" FT CDS complement(331..2802) FT /vntifkey="4" FT /label=hMLT FT mutation complement(2797..2799) FT /vntifkey="62" FT /label=Kozak FT /note="TCG was replaced by GCG (S2A) creating Kozak consensus sequence GCCACCATGG" SQ Sequence 8251 BP; 1985 A; 2155 C; 2031 G; 2080 t; ttgtcatgtt tgacagctta tcttcgatta 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 atctaagctc tttttcagaa attctgagcc tgtcagagaa 360 actaaatggt atttcatcag ttgtctctac tggcacatta cttctactaa aatgacatga 420 agcatggtga gcgaaacttg aaataaatgc atctggagtc cggctgcaat gacagctatc 480 tgctggtgta acattacttg gattaccagg atgtgaatgg taaacaccat ggaatgggtc 540 ttgcaatgag tgataatgcc ctgctcctcc tgaggtggct gcagaactct ggtaaggacc 600 attagacata agacaagttt gaaagcaagt cttccttccc aaacctcgat gcatgtctaa 660 tttagcaatg agaggtttcc caacattcac ttcctgcttg tcctctacag tatcttccaa 720 tcctgagtac tgatatgata aacatactgt gaagactaga tgttccttta atttttgcag 780 tgaactgagt ctggtataga ggcaatgctt gggaagatcc tttgatacca agtagctgcc 840 agtttcttca ggcgtgcctt tatttgcatc ttttggatca atatctagat caagtggaaa 900 atcagtaacg taggcatcac acattattat ctccggtggt ttgtaaacta tacttgtata 960 gatgatcatg acattggaaa actcagctgc aaatcctaat tgaatctgaa caccacagtc 1020 aaacttaaga cacatacttt ctggaagttc atgagccttg gcccactgta gattccgcac 1080 aagagattca gcagaatatt ctgttccctg tattggatca gtaagtgctc tcttctcaga 1140 taaactactt cgaatctcta gagcctgttt gcctttggta aggtgacact tacccatatc 1200 ttctgcaact tcatccagta acacagtgat tttcttatct tctaataatc tgtcttttaa 1260 aaatttcata aagattccat ttgccaatcc agaatgctgg atttcaaaag cttcagctcc 1320 ttgacacgtg gcatatccaa acacaatatt ggcggtgact tttagtgcat ccaagattgg 1380 aatggtatca tcgtagtcat ttcttttcct acacatatcc aataagaaca cattaagtcc 1440 agtttctttt tcttgcatca atttcagtat attttgtaca cacagacaat tttcagacct 1500 atatggattt ggagcatcaa cggggaccat gaagctgttc ccaaaatttt cataaccatg 1560 tcctgcataa tataataacc catatactcc cttgtctaaa aggagtaaaa actcatccac 1620 agcattacgc atctcatatt cagtaagatc caacagtgaa accactttga agtccagctg 1680 tctcagtaag ttagtcaatt cgtacacatc caccaaagga gctttgagct tggggtgctc 1740 ccggtaattc atatttccta tcaaaagggc aaccttgtcc ttcgccaaag gctggtcagt 1800 tgtttgctct ttattatcag gatgaccaag attatttaat tcatcttcag tgcactccac 1860 tgcctcatct gttcttccta tgatgatttc taccttcttg ctatcttgac tgtctcgatc 1920 attatataca tgacaccagt aggttccttg gtgttccaaa tccacataag gcaccatgta 1980 tagctttttg gtctcatgtg ttaatggtaa ttcatttttg aaccactggt agtgaggaat 2040 agggcttcca acagcaacac actgtaaaac caatgtgctg cctggcatca gcttttggga 2100 agttggttca acacagattt gcaacttgga ttcagagacg ccatcaacac ttctctggaa 2160 gctctctggg atgtcgcaaa catccagctg tgaccactgg ctgaattcaa aggtgaaatt 2220 gttattaact cgacagacat aaaagcctgc atcttttaca tgcactgcat taaaaataag 2280 ctctgatgta tttccatttg gaatctcttt attcattttg aaccactgat attgaacaaa 2340 aggatgtcca gttgcccggc aacacagttt cacaaactgt ccagccaaga ctgcctttga 2400 ctctgggttt acagtaatct ttattcctgg ggggctgaga agctgaagaa cttcagtgtg 2460 ttccatagcc tgcaggaaat cactcaattc tgtgactgtg caaccttttt cacccattaa 2520 cttcagcaga cacaggctgg ggcttccttc aggctccagt accttaagag aacactgctc 2580 caggtctagg caactgaggc ggaggcgccc gcgactcccc gccagctccg ccagtctcct 2640 ccagccccgg ccctcgggcg cctgatccag gagctcgctg agcctccgca gcagcggctc 2700 ccgcaggcgg ttgagggtcg cgccggccgg aggggcgagc agcggccccg tgggggcggc 2760 cgagggcggc agggcctgta gcgggtcccc caacagcgcc atggtggcag gatccatgga 2820 gtctagaatt cccggggatc cacgcggaac cagaagtgcc ccggaggatg agattttctt 2880 aaagcggttg gctgctgaga cggctatgaa attctttttc catcgtcgct tcatatgtat 2940 atctccttct taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat 3000 tcccctatag tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc tacgccggac 3060 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca 3120 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg 3180 gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat 3240 tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg 3300 agtcgcataa gggagagcgt cgagatcccg gacaccatcg aatggcgcaa aacctttcgc 3360 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta 3420 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg 3480 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag 3540 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt 3600 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa 3660 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc 3720 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt 3780 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg 3840 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa 3900 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg 3960 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat 4020 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc 4080 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt 4140 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt 4200 ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg 4260 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg 4320 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg 4380 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat 4440 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc 4500 aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc ccttgagagc 4560 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat 4620 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt 4680 cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg 4740 aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga 4800 gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt 4860 cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg 4920 gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct 4980 tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc actggaccgc tgatcgtcac 5040 ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc 5100 cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac 5160 ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat 5220 caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg 5280 tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc ctggccacgg 5340 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact 5400 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg 5460 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa 5520 acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg 5580 ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt 5640 tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac 5700 cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc 5760 attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa 5820 accgccctta acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc 5880 aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat 5940 gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg 6000 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt 6060 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc 6120 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 6180 accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 6240 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6300 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6360 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6420 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6480 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6540 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6600 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6660 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6720 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 6780 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6840 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 6900 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 6960 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7020 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7080 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7140 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7200 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7260 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7320 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7380 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7440 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc 7500 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7560 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7620 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7680 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7740 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac 7800 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7860 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 7920 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 7980 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8040 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8100 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8160 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8220 gcgtatcacg aggccctttc gtcttcaaga a 8251 //