ID pCal-n-A7M8K standard; circular DNA; ; 8617 BP. DE LMBP 5465. CC Accession number: LMBP 5465 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 CDS complement(331..1844) FT /vntifkey="4" FT /label=hMLT\fragment FT /note="exon 8 --> end" FT CDS complement(1845..3168) FT /vntifkey="4" FT /label=hAPI2\fragment FT /note="exon 1 --> 7" FT promoter 3697..3774 FT /vntifkey="30" FT /label=lacI\promoter FT /note="E. coli repressor lacI promoter, incl. ribosome binding site" FT RBS complement(3301..3345) FT /vntifkey="32" FT /label=5'\UTR\of\T7\gene\10,\incl.\RBS FT promoter complement(3347..3388) FT /vntifkey="30" FT /label=T7\promoter\with\lac\operator FT CDS 3775..4857 FT /vntifkey="4" FT /label=lacI FT CDS 6169..6360 FT /vntifkey="4" FT /label=rop FT /note="modulation repressor coding sequence" FT rep_origin complement(6606..7008) FT /vntifkey="33" FT /label=pMB1\ori FT CDS complement(7549..8409) FT /vntifkey="4" FT /label=ampicillin\resistance FT terminator complement(213..260) FT /vntifkey="43" FT /label=T7\terminator FT misc_marker complement(3202..3219) FT /vntifkey="22" FT /label=thrombin\recognition\site FT misc_marker complement(3220..3297) 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 mutation complement(1915..1917) FT /vntifkey="62" FT /label=PCR\artefact FT /note="silent mutation; TTA was replaced by TTG" FT mutation complement(3163..3165) FT /vntifkey="62" FT /label=Kozak FT /note="AAC was replaced by GAC (N2D) creating Kozak consensus sequence GCCACCATGG" SQ Sequence 8617 BP; 2126 A; 2181 C; 2068 G; 2242 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 tcatttgatt ctttttcctc 1860 agttgctctt tctctctcct cttcccttat ttcatcttct gcattgagta agtccaacac 1920 aagatcattg actagtctat aattctctcc agttgctagg atttttctct gaactgtctg 1980 ttttaccagg cttctactaa agcccatttc cacggcagca ttaatcacag gagtattcat 2040 catgattgca tcttctgaat ggtcttctcc aggttcaaaa tggataattg atgactctgc 2100 attttcatct cctgggctgt ctgatgtgga tagcagctgt tcaagtagat gagggtaact 2160 ggcttgaact tgacggatga actcctgtcc tttaattctt atcaagtact cacaccttgg 2220 aaaccacttg gcatgttgaa cccatggatc atctccagat tcccaacacc tgagtccacc 2280 atcacagcaa aagcatttga catcatcact gttacccaca taataaaaac ccgcacttgc 2340 aagctgctca ggattaacta gaacactaga gggccagtta aagaatgttt taaagcgggc 2400 tgcatgtgtc tgcatgctca gattagaaac tgtgtatctt gaagtgtctt gaagctgatt 2460 ttctataaat gggcatttgg gaaaatgtct caggtgttct gacatagcat tatccttcgg 2520 ttcccaattg ctcaattttc caccacaggc aaagcaagcc actctgtctc caggtcctat 2580 gtagtaaaag cctgcttttg ccagatctgt tggcgacaga aaagtcaatg gccatgtctg 2640 aaaagtaagt aatctggcat tttcgttatt cattgcacag tggtaggaac ttctcatcaa 2700 ggcagaaaaa tcttgatttg ctctggagtt tacaggattt gatggagagt ttgaataaga 2760 gccacggaaa tatccactgt tttctgtacc cggaagtaat gagtgtgtgg aatttgttac 2820 tgaagaagga aaagtaggct gagaggtagc ttccaagttg ttaacggaat ttagactctg 2880 aacgaatctg cagctaggat acaacttttt atgcttttca gtaggactgt ctcctctttt 2940 ccagttatcc agcatcaggc cacaacagaa gcatttgacc ttgtcattca caccagtgta 3000 atagaaacca gcacgagcaa gactcctttc tgagacagga accccagcag gaaaagtgga 3060 atacgtagac attcggtaca gttcacatga caagtcgtat ttcagttcaa acgtgttggc 3120 gcttttcatc aaatttgata agaatatgct gttttctact atgtccatgg tggcaggatc 3180 catggagtct agaattcccg gggatccacg cggaaccaga agtgccccgg aggatgagat 3240 tttcttaaag cggttggctg ctgagacggc tatgaaattc tttttccatc gtcgcttcat 3300 atgtatatct ccttcttaaa gttaaacaaa attatttcta gaggggaatt gttatccgct 3360 cacaattccc ctatagtgag tcgtattaat ttcgcgggat cgagatctcg atcctctacg 3420 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg 3480 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg 3540 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg 3600 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa 3660 tgcaggagtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc 3720 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa 3780 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc 3840 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg 3900 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg 3960 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg 4020 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc 4080 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg 4140 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat 4200 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc 4260 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc 4320 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat 4380 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc 4440 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg 4500 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg 4560 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat 4620 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac 4680 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca 4740 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg 4800 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg 4860 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt 4920 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc 4980 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt 5040 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt 5100 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt 5160 cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct acgtcttgct 5220 ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg cttccggcgg 5280 catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg 5340 acagcttcaa ggatcgctcg cggctcttac cagcctaact tcgatcactg gaccgctgat 5400 cgtcacggcg atttatgccg cctcggcgag cacatggaac gggttggcat ggattgtagg 5460 cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac 5520 ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccactcc aagaattgga 5580 gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc 5640 atcgcgtccg ccatctccag cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg 5700 ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc 5760 cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc 5820 aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt 5880 ctggaaacgc ggaagtcagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc 5940 tgctggctac cctgtggaac acctacatct gtattaacga agcgctggca ttgaccctga 6000 gtgatttttc tctggtcccg ccgcatccat accgccagtt gtttaccctc acaacgttcc 6060 agtaaccggg catgttcatc atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc 6120 ggtatcatta cccccatgaa cagaaatccc ccttacacgg aggcatcagt gaccaaacag 6180 gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag 6240 aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac 6300 gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga 6360 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa 6420 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca 6480 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga 6540 gagtgcacca tatatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat 6600 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 6660 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 6720 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 6780 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 6840 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 6900 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 6960 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7020 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7080 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7140 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7200 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7260 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7320 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7380 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7440 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7500 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7560 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7620 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 7680 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 7740 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 7800 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 7860 tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 7920 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 7980 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8040 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8100 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8160 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8220 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8280 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8340 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8400 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8460 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8520 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 8580 aaataggcgt atcacgaggc cctttcgtct tcaagaa 8617 //