LOCUS V029177 7621 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029177 VERSION V029177 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 964..3111 /label="Dvl3(NM_007889)" /note="Dvl3(NM_007889)" /gene="Dvl3" CDS 3118..3183 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3221..3445 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3491..3919 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3933..4262 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4329..5123 /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 5297..5418 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5467..5483) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5491..5507 /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(5515..5545) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5560..5581 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5869..6454) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6625..7485) /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(7486..7590) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatgggcg agaccaagat catctaccac ctggacggcc aggagacgcc gtacctggtg 1021 aagctgcccc tgcccgccga gcgcgtcacc ttggcggact ttaagggcgt tctgcagcgg 1081 cccagctata agttcttctt caagtctatg gacgacgatt ttggggtggt gaaggaagag 1141 atctcggacg acaatgccaa gctaccctgc ttcaatggcc gggtggtgtc ctggctggta 1201 tctgccgagg gctcacaccc agagccggcc cctttctgtg ctgacaaccc gtcagagctg 1261 ccaccttcga tggaacgcac aggaggcatt ggggactctc gacccccctc cttccaccct 1321 catgctagtg ggggcagcca ggagaatctg gacaatgaca cggagaccga ctccttggtg 1381 tctgcccaac gagagcggcc ccgacggagg gatggcccgg agcacgcagc ccgactcaat 1441 ggaaccacaa agggtgagcg gcgacgggag ccaggcggct atgacagttc atctaccctc 1501 atgagcagcg agctggagac taccagtttc tttgattccg atgaggatga ttccaccagc 1561 aggttcagca gctccacaga acagagcagt gcctcacgcc tcatgagaag acacaagcgg 1621 cggcggcgga agcagaaggt ctcccggatt gagcggtcct cgtctttcag cagcatcaca 1681 gactccacta tgtctctcaa catcatcacc gtcactctca acatggaaaa gtataacttc 1741 ttgggcatct ccattgtggg ccaaagcaat gagcgaggtg atgggggcat ctacattggc 1801 tccatcatga agggtggggc tgtggcagct gatggacgca ttgagccagg agatatgcta 1861 ctgcaggtaa atgagatcaa cttcgagaac atgagcaacg atgacgcagt ccgagtcctt 1921 cgggagattg tgcacaagcc agggcccatc actctgactg tagccaagtg ctgggaccca 1981 agtcctcgcg gctgcttcac attaccccgg agcgagccca tccgtcccat tgaccctgca 2041 gcctgggtct ctcacacggc agccatgaca ggcaccttcc ccgcctatgg catgagcccc 2101 tccttaagca ccatcacctc taccagctcc tccatcacca gctccatccc tgatacagag 2161 cgcctagatg atttccacct ctccatccac agtgacatgg cggccatcgt aaaagccatg 2221 gcctcccctg agtcagggtt ggaggtccga gaccgcatgt ggctcaagat taccattccc 2281 aatgctttca tcggctcgga tgtggtggac tggctgtacc acaacgtgga agggttcacc 2341 gaccgtcggg aggcccgcaa gtatgctagc aacctgctga aagctggttt catccgccat 2401 accgtcaaca agatcacgtt ctccgagcag tgctactaca tctttggcga cctctgtggt 2461 aacatggcca acctatcgct tcacgaccat gatggctcca gtggcgcctc tgaccaggac 2521 acactggctc ctttgccgca cccaggagct gccccttggc ctatggcttt cccttaccag 2581 tacccaccac ctccgcatcc ctacaaccca cacccaggct tcccagaact gggctacagc 2641 tacggtgggg gcagcgccag cagtcagcac agtgaaggca gtcggagcag tggctccaac 2701 cgaagtggca gcgaccggcg caaggagaag gacccaaaag ctggggactc caagtcaggt 2761 ggtagcggca gcgagtcaga ccacactaca cgcagcagtc tgagggggcc acgggagcgg 2821 gcaccaagcg agcgatcagg cccagcggcc agcgagcaca gccaccgcag tcaccattcg 2881 ctgaccagca gcctgcgcag ccaccacaca cacccaagct atggcccacc cggagtgccc 2941 cctctctatg gtcccccgat gctgatgatg accccgccgc cggcagccat gggtccccca 3001 ggagcccctc cagggcgtga cctggcctcg gtgcccccgg aactgacggc cagcagacag 3061 tccttccgca tggcaatggg aaaccccagt gagttctttg tggatgtgat gctcgaggac 3121 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 3181 aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag 3241 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 3301 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 3361 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 3421 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg 3481 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 3541 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 3601 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 3661 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 3721 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 3781 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 3841 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 3901 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 3961 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 4021 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 4081 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 4141 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 4201 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 4261 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat 4321 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 4381 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 4441 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 4501 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 4561 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 4621 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 4681 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 4741 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 4801 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca 4861 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 4921 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 4981 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 5041 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 5101 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 5161 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 5221 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 5281 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag 5341 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 5401 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta 5461 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 5521 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 5581 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 5641 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 5701 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 5761 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 5821 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 5881 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 5941 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 6001 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 6061 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 6121 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 6181 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 6241 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 6301 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 6361 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa 6421 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 6481 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 6541 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 6601 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 6661 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 6721 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 6781 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6841 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6901 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6961 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 7021 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 7081 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 7141 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 7201 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 7261 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 7321 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 7381 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 7441 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 7501 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 7561 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 7621 c //