ID pCD-F-HLH-MK-E4A-bio standard; circular DNA; ; 8432 BP. DE LMBP 5683. CC Accession number: LMBP 5683 CC (c) BCCM/GeneCorner Plasmid collection (Ghent University, Belgium) CC Last modified: 27-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 3950..3994 FT /vntifkey="22" FT /label=bio-tag FT mutation 971..973 FT /vntifkey="62" FT /label=Kozak FT /note="TCG was replaced by GCG (S2A) creating Kozak consensus sequence GCCACCATGG" FT mutation 3410..3412 FT /vntifkey="62" FT /label=PCR\artefact FT /note="silent mutation; wt: ACA" FT mutation 2948..2950 FT /vntifkey="62" FT /label=stability\creator FT /note="silent mutation; wt: GCA ('GCA'GAAG recombines with the same heptamer in hAPI2)" FT terminator 4029..4253 FT /vntifkey="43" FT /label=BGH\polyA FT rep_origin 4299..4727 FT /vntifkey="33" FT /label=f1\ori FT rep_origin 4970..5045 FT /vntifkey="33" FT /label=SV40\ori FT promoter 4732..5013 FT /vntifkey="29" FT /label=SV40\early\promoter FT CDS 5137..5931 FT /vntifkey="4" FT /label=Tn5\neomycin\resistance FT terminator 6105..6179 FT /vntifkey="43" FT /label=SV40\polyA\early FT terminator complement(6105..6179) FT /vntifkey="43" FT /label=SV40\polyA\late FT rep_origin complement(6493..6895) FT /vntifkey="33" FT /label=pMB1\ori FT CDS complement(7436..8296) FT /vntifkey="4" FT /label=ampicillin\resistance FT promoter complement(6318..6401) FT /vntifkey="30" FT /label=lac\promoter FT CDS 968..1447 FT /vntifkey="4" FT /label=HLH\dimerization\domain FT /note="(hMLT: 968-1093; hETV6: 1094-1447)" FT CDS 1463..3934 FT /vntifkey="4" FT /label=hMLT\mutant FT mutation 1466..1468 FT /vntifkey="62" FT /label=Kozak FT /note="TCG was replaced by GCG (S2A) creating Kozak consensus sequence GCCACCATGG" FT mutation 3419..3427 FT /vntifkey="62" FT /label=E653A FT /note="wt: GAA.ACT.GGC" FT mutation 3875..3880 FT /vntifkey="62" FT /label=E806A FT /note="wt: GTA.GAG" FT mutation 2399..2401 FT /vntifkey="62" FT /label=E313A FT /note="wt: GAG" FT mutation 2408..2410 FT /vntifkey="62" FT /label=E316A FT /note="wt: GAG" SQ Sequence 8432 BP; 2079 A; 2087 C; 2110 G; 2156 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 ctaaggatcc 960 tgccaccatg gcgctgttgg gggacccgct acaggccctg ccgccctcgg ccgcccccac 1020 ggggccgctg ctcgcccctc cggccggcgc gaccctcaac cgcctgcggg agccgctgct 1080 gcggaggctc agctacgctt cctcgacgcc acttcatgtt ccagtgcctc gagcgctcag 1140 gatggaggaa gactcgatcc gcctgcctgc gcacctgcgc ttgcagccaa tttactggag 1200 cagggatgac gtagcccagt ggctcaagtg ggctgaaaat gagttttctt taaggccaat 1260 tgacagcaac acgtttgaaa tgaatggcaa agctctcctg ctgctgacca aagaggactt 1320 tcgctatcga tctcctcatt caggtgatgt gctctatgaa ctccttcagc atattctgaa 1380 gcagaggaaa cctcggattc ttttttcacc attcttccac cctggaaact ctatacacac 1440 acagccggtt gatcctgcca ccatggcgct gttgggggac ccgctacagg ccctgccgcc 1500 ctcggccgcc cccacggggc cgctgctcgc ccctccggcc ggcgcgaccc tcaaccgcct 1560 gcgggagccg ctgctgcgga ggctcagcga gctcctggat caggcgcccg agggccgggg 1620 ctggaggaga ctggcggagc tggcggggag tcgcgggcgc ctccgcctca gttgcctaga 1680 cctggagcag tgttctctta aggtactgga gcctgaagga agccccagcc tgtgtctgct 1740 gaagttaatg ggtgaaaaag gttgcacagt cacagaattg agtgatttcc tgcaggctat 1800 ggaacacact gaagttcttc agcttctcag ccccccagga ataaagatta ctgtaaaccc 1860 agagtcaaag gcagtcttgg ctggacagtt tgtgaaactg tgttgccggg caactggaca 1920 tccttttgtt caatatcagt ggttcaaaat gaataaagag attccaaatg gaaatacatc 1980 agagcttatt tttaatgcag tgcatgtaaa agatgcaggc ttttatgtct gtcgagttaa 2040 taacaatttc acctttgaat tcagccagtg gtcacagctg gatgtttgcg acatcccaga 2100 gagcttccag agaagtgttg atggcgtctc tgaatccaag ttgcaaatct gtgttgaacc 2160 aacttcccaa aagctgatgc caggcagcac attggtttta cagtgtgttg ctgttggaag 2220 ccctattcct cactaccagt ggttcaaaaa tgaattacca ttaacacatg agaccaaaaa 2280 gctatacatg gtgccttatg tggatttgga acaccaagga acctactggt gtcatgtata 2340 taatgatcga gacagtcaag atagcaagaa ggtagaaatc atcataggaa gaacagatgc 2400 ggcagtggca tgcactgaag atgaattaaa taatcttggt catcctgata ataaagagca 2460 aacaactgac cagcctttgg cgaaggacaa ggttgccctt ttgataggaa atatgaatta 2520 ccgggagcac cccaagctca aagctccttt ggtggatgtg tacgaattga ctaacttact 2580 gagacagctg gacttcaaag tggtttcact gttggatctt actgaatatg agatgcgtaa 2640 tgctgtggat gagtttttac tccttttaga caagggagta tatgggttat tatattatgc 2700 aggacatggt tatgaaaatt ttgggaacag cttcatggtc cccgttgatg ctccaaatcc 2760 atataggtct gaaaattgtc tgtgtgtaca aaatatactg aaattgatgc aagaaaaaga 2820 aactggactt aatgtgttct tattggatat gtgtaggaaa agaaatgact acgatgatac 2880 cattccaatc ttggatgcac taaaagtcac cgccaatatt gtgtttggat atgccacgtg 2940 tcaaggagct gaagcttttg aaatccagca ttctggattg gcaaatggaa tctttatgaa 3000 atttttaaaa gacagattat tagaagataa gaaaatcact gtgttactgg atgaagttgc 3060 agaagatatg ggtaagtgtc accttaccaa aggcaaacag gctctagaga ttcgaagtag 3120 tttatctgag aagagagcac ttactgatcc aatacaggga acagaatatt ctgctgaatc 3180 tcttgtgcgg aatctacagt gggccaaggc tcatgaactt ccagaaagta tgtgtcttaa 3240 gtttgactgt ggtgttcaga ttcaattagg atttgcagct gagttttcca atgtcatgat 3300 catctataca agtatagttt acaaaccacc ggagataata atgtgtgatg cctacgttac 3360 tgattttcca cttgatctag atattgatcc aaaagatgca aataaaggca cgcctgaagc 3420 aaccggtagc tacttggtat caaaggatct tcccaagcat tgcctctata ccagactcag 3480 ttcactgcaa aaattaaagg aacatctagt cttcacagta tgtttatcat atcagtactc 3540 aggattggaa gatactgtag aggacaagca ggaagtgaat gttgggaaac ctctcattgc 3600 taaattagac atgcatcgag gtttgggaag gaagacttgc tttcaaactt gtcttatgtc 3660 taatggtcct taccagagtt ctgcagccac ctcaggagga gcagggcatt atcactcatt 3720 gcaagaccca ttccatggtg tttaccattc acatcctggt aatccaagta atgttacacc 3780 agcagatagc tgtcattgca gccggactcc agatgcattt atttcaagtt tcgctcacca 3840 tgcttcatgt cattttagta gaagtaatgt gccagtcgcg acaactgatg aaataccatt 3900 tagtttctct gacaggctca gaatttctga aaaagggccc gtttccggag gcctgaacga 3960 catcttcgag gcccagaaga tcgagtggca cgagtaaggg cccgtttaaa cccgctgatc 4020 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc 4080 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc 4140 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg 4200 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga 4260 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 4320 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 4380 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 4440 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 4500 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 4560 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 4620 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 4680 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 4740 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 4800 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 4860 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 4920 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 4980 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 5040 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc 5100 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 5160 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 5220 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 5280 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 5340 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 5400 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 5460 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 5520 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 5580 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 5640 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 5700 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 5760 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 5820 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 5880 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 5940 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 6000 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 6060 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag 6120 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 6180 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac 6240 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 6300 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 6360 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 6420 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 6480 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 6540 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 6600 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 6660 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 6720 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 6780 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 6840 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 6900 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 6960 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 7020 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 7080 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 7140 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 7200 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 7260 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 7320 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 7380 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 7440 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 7500 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 7560 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 7620 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 7680 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 7740 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 7800 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 7860 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 7920 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 7980 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 8040 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 8100 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 8160 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 8220 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 8280 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 8340 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 8400 gcacatttcc ccgaaaagtg ccacctgacg tc 8432 //