LOCUS V020155 7226 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020155 VERSION V020155 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 908..2662 /label="FZD5(NM_003468)" /note="FZD5(NM_003468)" /gene="FZD5" CDS 2669..2758 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" polyA_signal 2793..3017 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3063..3491 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3505..3834 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3901..4695 /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 4869..4990 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5039..5055) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5063..5079 /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(5087..5117) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5132..5153 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5441..6026) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6197..7057) /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(7058..7162) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cgccaccatg gctcggcctg acccatccgc gccgccctcg ctgttgctgc tgctcctagc 961 gcagctggtg ggccgggcgg ccgccgcgtc caaggccccg gtgtgccagg aaatcacggt 1021 gcccatgtgc cgcggcatcg gctacaacct gacgcacatg cccaaccagt tcaaccacga 1081 cacgcaggac gaggcgggcc tggaggtgca ccagttctgg ccgctggtgg agatccaatg 1141 ctcgccggac ctgcgcttct tcctatgctc tatgtacacg cccatctgtc tgcccgacta 1201 ccacaagccg ctgccgccct gccgctcggt gtgcgagcgc gccaaggccg gctgctcgcc 1261 gctgatgcgc cagtacggct tcgcctggcc cgagcgcatg agctgcgacc gcctcccggt 1321 gctgggccgc gacgccgagg tcctctgcat ggattacaac cgcagcgagg ccaccacggc 1381 gccccccagg cctttcccag ccaagcccac ccttccaggc ccgccagggg cgccggcctc 1441 ggggggcgaa tgccccgctg ggggcccgtt cgtgtgcaag tgtcgcgagc ccttcgtgcc 1501 cattctgaag gagtcacacc cgctctacaa caaggtgcgg acgggccagg tgcccaactg 1561 cgcggtaccc tgctaccagc cgtccttcag tgccgacgag cgcacgttcg ccaccttctg 1621 gataggcctg tggtcggtgc tgtgcttcat ctccacgtcc accacagtgg ccaccttcct 1681 catcgacatg gaacgcttcc gctatcctga gcgccccatc atcttcctgt cagcctgcta 1741 cctgtgcgtg tcgctgggct tcctggtgcg tctggtcgtg ggccatgcca gcgtggcctg 1801 cagccgcgag cacaaccaca tccactacga gaccacgggc cctgcactgt gcaccatcgt 1861 cttcctcctg gtctacttct tcggcatggc cagctccatc tggtgggtca tcctgtcgct 1921 cacctggttc ctggccgccg gcatgaagtg gggcaacgag gccatcgcgg gctacgcgca 1981 gtacttccac ctggctgcgt ggctcatccc cagcgtcaag tccatcacgg cactggcgct 2041 gagctccgtg gacggggacc cagtggccgg catctgctac gtgggcaacc agaacctgaa 2101 ctcgctgcgc ggcttcgtgc tgggcccgct ggtgctctac ctgctggtgg gcacgctctt 2161 cctgctggcg ggcttcgtgt cgctcttccg catccgcagc gtcatcaagc agggcggcac 2221 caagacggac aagctggaga agctcatgat ccgcatcggc atcttcacgc tgctctacac 2281 ggtccccgcc agcattgtgg tggcctgcta cctgtacgag cagcactacc gcgagagctg 2341 ggaggcggcg ctcacctgcg cctgcccggg ccacgacacc ggccagccgc gcgccaagcc 2401 cgagtactgg gtgctcatgc tcaagtactt catgtgcctg gtggtgggca tcacgtcggg 2461 cgtctggatc tggtcgggca agacggtgga gtcgtggcgg cgtttcacca gccgctgctg 2521 ctgccgcccg cggcgcggcc acaagagcgg gggcgccatg gccgcagggg actaccccga 2581 ggcgagcgcc gcgctcacag gcaggaccgg gccgccgggc cccgccgcca cctaccacaa 2641 gcaggtgtcc ctgtcgcacg tgctcgagga acaaaaacta atatcagagg aagatttgga 2701 acagaaacta atcagtgaag aagatttaga gcaaaagtta atttccgagg aggacttatg 2761 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt 2821 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc 2881 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg 2941 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga 3001 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc 3061 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 3121 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 3181 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 3241 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 3301 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 3361 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 3421 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 3481 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 3541 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 3601 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 3661 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 3721 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct 3781 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc 3841 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc 3901 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc 3961 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca 4021 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg 4081 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg 4141 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag 4201 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg 4261 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc 4321 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa 4381 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac 4441 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat 4501 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac 4561 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc 4621 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt 4681 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc 4741 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 4801 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 4861 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 4921 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 4981 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 5041 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 5101 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 5161 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 5221 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 5281 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 5341 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 5401 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 5461 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 5521 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 5581 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 5641 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 5701 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 5761 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 5821 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 5881 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 5941 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga 6001 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 6061 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 6121 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 6181 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 6241 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 6301 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 6361 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 6421 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 6481 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 6541 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 6601 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 6661 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 6721 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 6781 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 6841 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 6901 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 6961 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 7021 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 7081 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 7141 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat 7201 cgggagatct cccgatcccc tatggt //