ID pCD-F-bio-MK-E4A standard; circular DNA; ; 7919 BP. DE LMBP 5680. CC Accession number: LMBP 5680 CC (c) BCCM/GeneCorner Plasmid collection (Ghent University, Belgium) CC Last modified: 31-JUL-2009 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 misc_marker 923..946 FT /vntifkey="22" FT /label=FLAG\epitope FT promoter 235..818 FT /vntifkey="29" FT /label=hCMV-IE\promoter\and\enhancer FT promoter 863..879 FT /vntifkey="30" FT /label=T7\promoter FT misc_marker 953..997 FT /vntifkey="22" FT /label=bio-tag FT CDS 1013..3484 FT /vntifkey="4" FT /label=hMLT\mutant FT mutation 1016..1018 FT /vntifkey="62" FT /label=Kozak FT /note="TCG was replaced by GCG (S2A) creating Kozak consensus sequence GCCACCATGG" FT mutation 2960..2962 FT /vntifkey="62" FT /label=PCR\artefact FT /note="silent mutation; wt: ACA" FT mutation 2498..2500 FT /vntifkey="62" FT /label=stability\creator FT /note="silent mutation; wt: GCA ('GCA'GAAG recombines with the same heptamer in hAPI2)" FT terminator 3516..3740 FT /vntifkey="43" FT /label=BGH\polyA FT rep_origin 3786..4214 FT /vntifkey="33" FT /label=f1\ori FT rep_origin 4457..4532 FT /vntifkey="33" FT /label=SV40\ori FT promoter 4219..4500 FT /vntifkey="29" FT /label=SV40\early\promoter FT CDS 4624..5418 FT /vntifkey="4" FT /label=Tn5\neomycin\resistance FT terminator 5592..5666 FT /vntifkey="43" FT /label=SV40\polyA\early FT terminator complement(5592..5666) FT /vntifkey="43" FT /label=SV40\polyA\late FT rep_origin complement(5980..6382) FT /vntifkey="33" FT /label=pMB1\ori FT CDS complement(6923..7783) FT /vntifkey="4" FT /label=ampicillin\resistance FT promoter complement(5805..5888) FT /vntifkey="30" FT /label=lac\promoter FT mutation 1949..1951 FT /vntifkey="62" FT /label=E313A FT /note="wt: GAG" FT mutation 1958..1960 FT /vntifkey="62" FT /label=E316A FT /note="wt: GAG" FT mutation 3425..3430 FT /vntifkey="62" FT /label=E806A FT /note="wt: GTA.GAG" FT mutation 2969..2977 FT /vntifkey="62" FT /label=E653A FT /note="wt: GAA.ACT.GGC" SQ Sequence 7919 BP; 1978 A; 1921 C; 1978 G; 2042 t; gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 60 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 120 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 180 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 240 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 300 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 360 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 420 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 480 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 540 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 600 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 660 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 720 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 780 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 840 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 900 gtttaaactt aagcttacca tggattacaa ggacgatgac gataaggtac caggcctgaa 960 cgacatcttc gaggcccaga agatcgagtg gcacgaggag gatcctgcca ccatggcgct 1020 gttgggggac ccgctacagg ccctgccgcc ctcggccgcc cccacggggc cgctgctcgc 1080 ccctccggcc ggcgcgaccc tcaaccgcct gcgggagccg ctgctgcgga ggctcagcga 1140 gctcctggat caggcgcccg agggccgggg ctggaggaga ctggcggagc tggcggggag 1200 tcgcgggcgc ctccgcctca gttgcctaga cctggagcag tgttctctta aggtactgga 1260 gcctgaagga agccccagcc tgtgtctgct gaagttaatg ggtgaaaaag gttgcacagt 1320 cacagaattg agtgatttcc tgcaggctat ggaacacact gaagttcttc agcttctcag 1380 ccccccagga ataaagatta ctgtaaaccc agagtcaaag gcagtcttgg ctggacagtt 1440 tgtgaaactg tgttgccggg caactggaca tccttttgtt caatatcagt ggttcaaaat 1500 gaataaagag attccaaatg gaaatacatc agagcttatt tttaatgcag tgcatgtaaa 1560 agatgcaggc ttttatgtct gtcgagttaa taacaatttc acctttgaat tcagccagtg 1620 gtcacagctg gatgtttgcg acatcccaga gagcttccag agaagtgttg atggcgtctc 1680 tgaatccaag ttgcaaatct gtgttgaacc aacttcccaa aagctgatgc caggcagcac 1740 attggtttta cagtgtgttg ctgttggaag ccctattcct cactaccagt ggttcaaaaa 1800 tgaattacca ttaacacatg agaccaaaaa gctatacatg gtgccttatg tggatttgga 1860 acaccaagga acctactggt gtcatgtata taatgatcga gacagtcaag atagcaagaa 1920 ggtagaaatc atcataggaa gaacagatgc ggcagtggca tgcactgaag atgaattaaa 1980 taatcttggt catcctgata ataaagagca aacaactgac cagcctttgg cgaaggacaa 2040 ggttgccctt ttgataggaa atatgaatta ccgggagcac cccaagctca aagctccttt 2100 ggtggatgtg tacgaattga ctaacttact gagacagctg gacttcaaag tggtttcact 2160 gttggatctt actgaatatg agatgcgtaa tgctgtggat gagtttttac tccttttaga 2220 caagggagta tatgggttat tatattatgc aggacatggt tatgaaaatt ttgggaacag 2280 cttcatggtc cccgttgatg ctccaaatcc atataggtct gaaaattgtc tgtgtgtaca 2340 aaatatactg aaattgatgc aagaaaaaga aactggactt aatgtgttct tattggatat 2400 gtgtaggaaa agaaatgact acgatgatac cattccaatc ttggatgcac taaaagtcac 2460 cgccaatatt gtgtttggat atgccacgtg tcaaggagct gaagcttttg aaatccagca 2520 ttctggattg gcaaatggaa tctttatgaa atttttaaaa gacagattat tagaagataa 2580 gaaaatcact gtgttactgg atgaagttgc agaagatatg ggtaagtgtc accttaccaa 2640 aggcaaacag gctctagaga ttcgaagtag tttatctgag aagagagcac ttactgatcc 2700 aatacaggga acagaatatt ctgctgaatc tcttgtgcgg aatctacagt gggccaaggc 2760 tcatgaactt ccagaaagta tgtgtcttaa gtttgactgt ggtgttcaga ttcaattagg 2820 atttgcagct gagttttcca atgtcatgat catctataca agtatagttt acaaaccacc 2880 ggagataata atgtgtgatg cctacgttac tgattttcca cttgatctag atattgatcc 2940 aaaagatgca aataaaggca cgcctgaagc aaccggtagc tacttggtat caaaggatct 3000 tcccaagcat tgcctctata ccagactcag ttcactgcaa aaattaaagg aacatctagt 3060 cttcacagta tgtttatcat atcagtactc aggattggaa gatactgtag aggacaagca 3120 ggaagtgaat gttgggaaac ctctcattgc taaattagac atgcatcgag gtttgggaag 3180 gaagacttgc tttcaaactt gtcttatgtc taatggtcct taccagagtt ctgcagccac 3240 ctcaggagga gcagggcatt atcactcatt gcaagaccca ttccatggtg tttaccattc 3300 acatcctggt aatccaagta atgttacacc agcagatagc tgtcattgca gccggactcc 3360 agatgcattt atttcaagtt tcgctcacca tgcttcatgt cattttagta gaagtaatgt 3420 gccagtcgcg acaactgatg aaataccatt tagtttctct gacaggctca gaatttctga 3480 aaaagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 3540 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 3600 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 3660 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 3720 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 3780 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 3840 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 3900 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 3960 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 4020 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 4080 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 4140 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 4200 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 4260 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 4320 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 4380 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 4440 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 4500 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 4560 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 4620 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 4680 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 4740 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 4800 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 4860 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 4920 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 4980 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 5040 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 5100 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 5160 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 5220 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 5280 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 5340 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 5400 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 5460 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 5520 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 5580 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 5640 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 5700 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 5760 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 5820 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 5880 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 5940 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 6000 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 6060 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 6120 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 6180 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 6240 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 6300 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 6360 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 6420 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 6480 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 6540 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 6600 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 6660 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 6720 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 6780 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 6840 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 6900 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 6960 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 7020 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 7080 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 7140 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 7200 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 7260 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 7320 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 7380 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 7440 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 7500 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 7560 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 7620 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 7680 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 7740 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 7800 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 7860 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc 7919 //