LOCUS V019031 6587 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019031 VERSION V019031 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 683..1267 /label="CYBA(NM_000101)" /note="CYBA(NM_000101)" /gene="CYBA" CDS 1274..1339 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1363..1951 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2017..2065 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2267..2695 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2709..3038 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3105..3899 /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 4075..4196 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4245..4261) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4269..4285 /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(4293..4323) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4338..4359 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4647..5235) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5406..6266) /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(6267..6371) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccgcca ccatggggca gatcgagtgg gccatgtggg ccaacgaaca 721 ggcgctggcg tccggcctga tcctcatcac cgggggcatc gtggccacag ctgggcgctt 781 cacccagtgg tactttggtg cctactccat tgtggcgggc gtgtttgtgt gcctgctgga 841 gtacccccgg gggaagagga agaagggctc caccatggag cgctggggac agaagtacat 901 gaccgccgtg gtgaagctgt tcgggccctt taccaggaat tactatgttc gggccgtcct 961 gcatctcctg ctctcggtgc ccgccggctt cctgctggcc accatccttg ggaccgcctg 1021 cctggccatt gcgagcggca tctacctact ggcggctgtg cgtggcgagc agtggacgcc 1081 catcgagccc aagccccggg agcggccgca gatcggaggc accatcaagc agccgcccag 1141 caaccccccg ccgcggcccc cggccgaggc ccgcaagaag cccagcgagg aggaggctgc 1201 ggtggcggcg gggggacccc cgggaggtcc ccaggtcaac cccatcccgg tgaccgacga 1261 ggtcgtgctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta 1321 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac 1381 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 1441 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 1501 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 1561 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 1621 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 1681 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 1741 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg 1801 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 1861 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 1921 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac 1981 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa 2041 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct 2101 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca 2161 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc 2221 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc 2281 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 2341 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 2401 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac 2461 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 2521 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 2581 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg 2641 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 2701 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 2761 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 2821 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 2881 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 2941 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3001 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 3061 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 3121 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 3181 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 3241 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 3301 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 3361 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 3421 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 3481 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 3541 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 3601 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 3661 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 3721 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 3781 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 3841 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 3901 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 3961 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 4021 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg 4081 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 4141 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 4201 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct 4261 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 4321 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 4381 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 4441 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 4501 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 4561 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 4621 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 4681 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 4741 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 4801 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat 4861 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 4921 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 4981 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 5041 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 5101 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 5161 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 5221 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 5281 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 5341 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 5401 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 5461 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 5521 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 5581 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 5641 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 5701 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 5761 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 5821 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 5881 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 5941 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 6001 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 6061 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 6121 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 6181 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 6241 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 6301 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 6361 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc 6421 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt 6481 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta 6541 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg //