LOCUS V018297 8453 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018297 VERSION V018297 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 941..3934 /label="GTF2I(NM_032999)" /note="GTF2I(NM_032999)" /gene="GTF2I" CDS 3950..4015 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 4053..4277 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4323..4751 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4765..5094 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5161..5955 /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 6129..6250 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6299..6315) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6323..6339 /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(6347..6377) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6392..6413 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6701..7286) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7457..8317) /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(8318..8422) /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 gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggcccaag ttgcaatgtc 961 caccctcccc gttgaagatg aggagtcctc ggagagcagg atggtggtga cattcctcat 1021 gtcagctctc gagtccatgt gtaaagaact ggccaagtcc aaagccgaag tggcctgcat 1081 tgcagtgtat gaaacagacg tgtttgtcgt cggaactgaa agaggacgtg cttttgtcaa 1141 taccagaaag gattttcaaa aagattttgt aaaatattgt gttgaagaag aagaaaaagc 1201 tgcagagatg cataaaatga aatctacaac ccaggcaaat cggatgagtg tagatgctgt 1261 agaaattgaa acactcagaa aaacagttga ggactatttc tgcttttgct atgggaaagc 1321 tttaggcaaa tccacagtgg tacctgtacc atatgagaag atgctgcgag accagtcggc 1381 tgtggtagtg caggggcttc cggaaggtgt tgcctttaaa caccccgaga actatgatct 1441 tgcaaccctg aaatggattt tggagaacaa agcagggatt tcattcatca ttaagagacc 1501 ttttttagag ccaaagaagc atgtaggtgg tcgtgtgatg gtaacagatg ctgacaggtc 1561 aatactatct ccaggtggaa gttgtggccc catcaaagtg aaaactgaac ccacagaaga 1621 ttctggtatt tccctggaaa tggcagctgt gacagtaaag gaagaatcag aagatcctga 1681 ttattatcaa tataacattc aagcaggccc ttctgaaact gatgatgttg atgaaaaaca 1741 gcccctatcg aagcctttgc aaggaagcca ccattcttca gagggcaatg aaggcacaga 1801 aatggaagta ccagcagaag attctactca acatgtccct tcagaaacaa gtgaggaccc 1861 tgaagttgag gtgactattg aagatgatga ttattctcca ccgtctaaga gaccaaaggc 1921 caatgagcta ccgcagccac cagtcccgga acccgccaat gctgggaagc ggaaagtgag 1981 ggagttcaac ttcgagaaat ggaatgctcg catcactgat ctacgtaaac aagttgaaga 2041 attgtttgaa aggaaatatg ctcaagccat aaaagccaaa ggtccggtga cgatcccgta 2101 ccctcttttc cagtctcatg ttgaagatct ttatgtagaa ggacttcctg aaggaattcc 2161 ttttagaagg ccatctactt acggaattcc tcgcctggag aggatattac ttgcaaagga 2221 aaggattcgt tttgtgatta agaaacatga gcttctgaat tcaacacgtg aagatttaca 2281 gcttgataag ccagcttcag gagtaaagga agaatggtat gccagaatca ctaaattaag 2341 aaagatggtg gatcagcttt tctgcaaaaa atttgcggaa gccttgggga gcactgaagc 2401 caaggctgta ccgtaccaaa aatttgaggc acacccgaat gatctgtacg tggaaggact 2461 gccagaaaac attcctttcc gaagtccctc atggtatgga atcccaaggc tggaaaaaat 2521 cattcaagtg ggcaatcgaa ttaaatttgt tattaaaaga ccagaacttc tgactcacag 2581 taccactgaa gttactcagc caagaacgaa tacaccagtc aaagaagatt ggaatgtcag 2641 aattaccaag ctacggaagc aagtggaaga gatttttaat ttgaaatttg ctcaagctct 2701 tggactcacc gaggcagtaa aagtaccata tcctgtgttt gaatcaaacc cggagttctt 2761 gtatgtggaa ggcttgccag aggggattcc cttccgaagc cctacctggt ttggaattcc 2821 acgacttgaa aggatcgtcc gcgggagtaa taaaatcaag ttcgttgtta aaaaacctga 2881 actagttatt tcctacttgc ctcctgggat ggctagtaaa ataaacacta aagctttgca 2941 gtcccccaaa agaccacgaa gtcctgggag taattcaaag gttcctgaaa ttgaggtcac 3001 cgtggaaggc cctaataaca acaatcctca aacctcagct gttcgaaccc cgacccagac 3061 taacggttct aacgttccct tcaagccacg agggagagag ttttcctttg aggcctggaa 3121 tgccaaaatc acggacctaa aacagaaagt tgaaaatctc ttcaatgaga aatgtgggga 3181 agctcttggc cttaaacaag ctgtgaaggt gccgttcgcg ttatttgagt ctttcccgga 3241 agacttttat gtggaaggct tacctgaggg tgtgccattc cgaagaccat cgacttttgg 3301 cattccgagg ctggagaaga tactcagaaa caaagccaaa attaagttca tcattaaaaa 3361 gcccgaaatg tttgagacgg cgattaagga gagcacctcc tctaagagcc ctcccagaaa 3421 aataaattca tcacccaatg ttaatactac tgcatcaggt gttgaagacc ttaacatcat 3481 tcaggtgaca attccagatg atgataatga aagactctcg aaagttgaaa aagctagaca 3541 gctaagagaa caagtgaatg acctctttag tcggaaattt ggtgaagcta ttggtatggg 3601 ttttcctgtg aaagttccct acaggaaaat cacaattaac cctggctgtg tggtagttga 3661 tggcatgccc ccgggggtgt ccttcaaagc ccccagctac ctggaaatca gctccatgag 3721 aaggatctta gactctgccg agtttatcaa attcacggtc attagaccat ttccaggact 3781 tgtgattaat aaccagctgg ttgatcagag tgagtcagaa ggccccgtga tacaagaatc 3841 agctgaacca agccagttgg aagttccagc cacagaagaa ataaaagaga ctgatggaag 3901 ctctcagatc aagcaagaac cagaccccac gtggatgcgg ccgctcgagg actacaaaga 3961 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg 4021 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt 4081 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc 4141 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg 4201 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga 4261 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc 4321 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 4381 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 4441 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 4501 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 4561 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 4621 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 4681 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 4741 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 4801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 4861 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 4921 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 4981 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct 5041 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc 5101 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc 5161 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc 5221 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca 5281 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg 5341 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg 5401 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag 5461 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg 5521 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc 5581 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa 5641 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac 5701 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat 5761 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac 5821 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc 5881 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt 5941 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc 6001 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 6061 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 6121 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6181 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6241 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 6301 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 6361 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 6421 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 6481 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 6541 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 6601 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 6661 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 6721 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 6781 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 6841 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 6901 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 6961 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7021 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7081 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7141 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7201 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga 7261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 7321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 7381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 7441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 7501 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 7561 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 7621 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 7681 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 7741 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 7801 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 7861 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 7921 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 7981 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 8041 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 8101 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 8161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 8221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 8281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 8341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 8401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc //