LOCUS V045333 7432 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045333 VERSION V045333 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 932..2911 /label="FOXK2(NM_004514)" /note="FOXK2(NM_004514)" /gene="FOXK2" CDS 2933..2998 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3027..3251 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3297..3725 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3739..4068 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4135..4929 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 5105..5226 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5275..5291) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5299..5315 /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(5323..5353) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5368..5389 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5677..6265) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6436..7296) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(7297..7401) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt 901 taagcttggt accgagctcg gatccgccac catggcagct gccgctgcag cattgtctgg 961 agctggaact cctccagctg gaggtggtgc tggtggtgga ggagcaggag gaggaggatc 1021 tcctccagga ggatgggcag tggcaagact cgagggccgc gagttcgagt atctgatgaa 1081 gaagcgctcg gtgaccatcg gccgcaactc gtcgcagggc tcggtggacg tgagcatggg 1141 ccactcgagc ttcatctccc ggcgccacct cgagatcttc acgcccccgg gcggcggcgg 1201 ccatggcggg gccgctccgg agctgccgcc cgcgcagccc aggcccgacg ccggcggcga 1261 cttctacctg cgctgcttgg gcaagaacgg ggtattcgtg gacggcgtgt tccagaggcg 1321 cggggcgccg ccgctgcagc tgccgcgcgt gtgcacattc aggttcccga gcacaaacat 1381 caagataacg ttcactgccc tgtccagcga gaagagagag aagcaggagg cgtctgagtc 1441 tccagtgaag gccgtacagc cacacatctc gcccctgacc atcaacattc cagacaccat 1501 ggcccacctc atcagccctc tgccctcccc cacgggaacc atcagcgctg caaactcctg 1561 cccctccagc ccccggggag cggggtcttc agggtacaag gtgggccgag tgatgccatc 1621 tgacctcaat ttaatggctg acaactcaca gcctgaaaat gaaaaggaag cttcaggtgg 1681 agacagcccg aaggatgatt caaagccgcc ttactcctac gcgcagctga tagttcaggc 1741 gattacgatg gctcccgaca aacagctcac cctgaacggg atttatacac acatcactaa 1801 aaattatccc tactacagga ctgcggacaa gggctggcag aattcaattc gccacaatct 1861 ctctctgaat cgttatttca tcaaagtgcc gcgttcccag gaagaaccag gcaaaggctc 1921 gttctggagg atagacccag cctctgaaag caaattaata gaacaggctt ttaggaaacg 1981 acggcctagg ggcgtgccct gctttagaac ccctctggga ccgctctctt ctaggagtgc 2041 cccagcctct cccaatcacg cgggagtgct gtctgctcac tctagtggcg cccagacccc 2101 tgagagcctg tcgagggaag gttcgccggc ccccctggag cctgagcctg gcgctgcaca 2161 gcccaaactc gctgtcatcc aggaagcccg gtttgcccag agcgccccag ggtcacctct 2221 gtccagtcag ccagtcttaa tcaccgtcca gcggcagcta ccacaggcca tcaagcctgt 2281 cacctacact gtggccaccc cagtgaccac ctcgacctcc cagccacccg tcgtgcagac 2341 ggttcacgtc gtccaccaga tcccagcggt gtcggtcacc agtgtggccg gactggcccc 2401 agcgaacacg tacactgtct ctggacaagc tgtggtcacc ccggcagccg tgctggcccc 2461 tcctaaggca gaggcccagg agaatggaga ccacagggaa gtcaaagtga aagtagagcc 2521 tattcccgcc attggccacg ccacgctcgg cactgccagc cggatcattc agacggcaca 2581 gaccaccccg gtccagacgg tgaccatagt acaacaggca cctctaggtc aacaccagct 2641 accaataaaa actgtaacac aaaacggcac tcacgtggca tcagtcccca ctgcggtcca 2701 cggccaggtg aacaatgccg cggcgagtcc tttgcacatg ttggcaacac acgcatccgc 2761 atcggcctcc ctgcccacaa agcgccacaa cggtgaccag ccggagcagc cggagctgaa 2821 gcggatcaag acagaagacg gcgagggcat cgtcattgcc ctgagcgtgg acacgccacc 2881 ggcagccgta agggaaaagg gtgtccagaa cctcgagtct agagggccct tcgactacaa 2941 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagtg 3001 agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 3061 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 3121 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 3181 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 3241 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 3301 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 3361 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 3421 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 3481 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 3541 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 3601 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 3661 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 3721 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 3781 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 3841 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 3901 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 3961 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta 4021 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga 4081 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 4141 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 4201 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 4261 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 4321 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 4381 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 4441 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 4501 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 4561 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 4621 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag 4681 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 4741 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 4801 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 4861 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 4921 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc 4981 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 5041 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 5101 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 5161 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 5221 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 5281 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 5341 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 5401 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 5461 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 5521 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 5581 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 5641 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 5701 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 5761 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 5821 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 5881 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 5941 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 6001 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 6061 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 6121 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 6181 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 6241 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 6301 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 6361 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 6421 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 6481 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 6541 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 6601 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 6661 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 6721 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 6781 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 6841 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 6901 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 6961 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 7021 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 7081 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 7141 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 7201 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 7261 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 7321 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 7381 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //