LOCUS V043038 5957 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043038 VERSION V043038 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 907..1404 /label="CFL1(NM_005507)" /note="CFL1(NM_005507)" /gene="CFL1" CDS 1414..1455 /codon_start=1 /product="epitope tag from simian virus 5" /transl_table=1 /translation="GKPIPNPLLGLDST" /label="V5 tag" polyA_signal 1557..1781 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1827..2255 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2269..2598 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2665..3459 /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 3633..3754 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3803..3819) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3827..3843 /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(3851..3881) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3896..3917 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4205..4790) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4961..5821) /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(5822..5926) /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 gccaccatgg cctccggtgt ggctgtctct gatggtgtca tcaaggtgtt caacgacatg 961 aaggtgcgta agtcttcaac gccagaggag gtgaagaagc gcaagaaggc ggtgctcttc 1021 tgcctgagtg aggacaagaa gaacatcatc ctggaggagg gcaaggagat cctggtgggc 1081 gatgtgggcc agactgtcga cgacccctac gccacctttg tcaagatgct gccagataag 1141 gactgccgct atgccctcta tgatgcaacc tatgagacca aggagagcaa gaaggaggat 1201 ctggtgttta tcttctgggc ccccgagtct gcgcccctta agagcaaaat gatttatgcc 1261 agctccaagg acgccatcaa gaagaagctg acagggatca agcatgaatt gcaagcaaac 1321 tgctacgagg aggtcaagga ccgctgcacc ctggcagaga agctgggggg cagtgccgtc 1381 atctccctgg agggcaagcc tttgaagctt atgggtaagc ctatccctaa ccctctcctc 1441 ggtctcgatt ctacatagga tccactagtc cagtgtggtg gaattctgca gatatccagc 1501 acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt 1561 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 1621 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 1681 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 1741 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 1801 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 1861 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 1921 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 1981 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 2041 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 2101 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 2161 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 2221 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 2281 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 2341 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 2401 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 2461 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 2521 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 2581 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 2641 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 2701 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 2761 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 2821 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 2881 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 2941 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 3001 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 3061 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 3121 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 3181 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 3241 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 3301 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 3361 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 3421 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 3481 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 3541 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 3601 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 3661 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 3721 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 3781 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 3841 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 3901 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 3961 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 4021 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 4081 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 4141 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 4201 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 4261 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 4321 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 4381 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 4441 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 4501 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 4561 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 4621 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 4681 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 4741 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 4801 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 4861 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 4921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 4981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 5041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 5101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 5161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 5221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 5281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 5341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 5401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 5461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 5521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 5581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 5641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 5701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 5761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 5821 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 5881 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 5941 aagtgccacc tgacgtc //