LOCUS V030196 8168 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030196 VERSION V030196 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..3658 /label="CTNNA1(NM_001903)" /note="CTNNA1(NM_001903)" /gene="CTNNA1" CDS 3665..3730 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3768..3992 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4038..4466 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4480..4809 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4876..5670 /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 5844..5965 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6014..6030) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6038..6054 /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(6062..6092) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6107..6128 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6416..7001) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7172..8032) /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(8033..8137) /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 atgactgctg tccatgcagg 961 caacataaac ttcaagtggg atcctaaaag tctagagatc aggactctgg cagttgagag 1021 actgttggag cctcttgtta cacaggttac aacccttgta aacaccaata gtaaagggcc 1081 ctctaataag aagagaggtc gttctaagaa ggcccatgtt ttggctgcat ctgttgaaca 1141 agcaactgag aatttcttgg agaaggggga taaaattgcg aaggagagcc agtttctcaa 1201 ggaggagctt gtggctgctg tagaagatgt tcgaaaacaa ggtgatttga tgaaggctgc 1261 tgcaggagag ttcgcagatg atccctgctc ttctgtgaag cgaggcaaca tggttcgggc 1321 agctcgagct ttgctctctg ctgttacccg gttgctgatt ttggctgaca tggcagatgt 1381 ctacaaatta cttgttcagc tgaaagttgt ggaagatggt atcttgaagt tgaggaatgc 1441 tggcaatgaa caagacttag gaatccagta taaagcccta aaacctgaag tggataagct 1501 gaacattatg gcagccaaaa gacaacagga attgaaagat gttggccatc gtgatcagat 1561 ggctgcagct agaggaatcc tgcagaagaa cgttccgatc ctctatactg catcccaggc 1621 atgcctacag caccctgatg tcgcagccta taaggccaac agggacctga tatacaagca 1681 gctgcagcag gcggtcacag gcatttccaa tgcagcccag gccactgcct cagacgatgc 1741 ctcacagcac cagggtggag gaggaggaga actggcatat gcactcaata actttgacaa 1801 acaaatcatt gtggacccct tgagcttcag cgaggagcgc tttaggcctt ccctggagga 1861 gcgtctggaa agcatcatta gtggggctgc cttgatggcc gactcgtcct gcacgcgtga 1921 tgaccgtcgt gagcgaattg tggcagagtg taatgctgtc cgccaggccc tgcaggacct 1981 gctttcggag tacatgggca atgctggacg taaagaaaga agtgatgcac tcaattctgc 2041 aatagataaa atgaccaaga agaccaggga cttgcgtaga cagctccgca aagctgtcat 2101 ggaccacgtt tcagattctt tcctggaaac caatgttcca cttttggtat tgattgaagc 2161 tgcaaagaat ggaaatgaga aagaagttaa ggagtatgcc caagttttcc gtgaacatgc 2221 caacaaattg attgaggttg ccaacttggc ctgttccatc tcaaataatg aagaaggtgt 2281 aaagcttgtt cgaatgtctg caagccagtt agaagccctc tgtcctcagg ttattaatgc 2341 tgcactggct ttagcagcaa aaccacagag taaactggcc caagagaaca tggatctttt 2401 taaagaacaa tgggaaaaac aagtccgtgt tctcacagat gctgtcgatg acattacttc 2461 cattgatgac ttcttggctg tctcagagaa tcacattttg gaagatgtga acaaatgtgt 2521 cattgctctc caagagaagg atgtggatgg cctggaccgc acagctggtg caattcgagg 2581 ccgggcagcc cgggtcattc acgtagtcac ctcagagatg gacaactatg agccaggagt 2641 ctacacagag aaggttctgg aagccactaa gctgctctcc aacacagtca tgccacgttt 2701 tactgagcaa gtagaagcag ccgtggaagc cctcagctcg gaccctgccc agcccatgga 2761 tgagaatgag tttatcgatg cttcccgcct ggtatatgat ggcatccggg acatcaggaa 2821 agcagtgctg atgataagga cccctgagga gttggatgac tctgactttg agacagaaga 2881 ttttgatgtc agaagcagga cgagcgtcca gacagaagac gatcagctga tagctggcca 2941 gagtgcccgg gcgatcatgg ctcagcttcc ccaggagcaa aaagcgaaga ttgcggaaca 3001 ggtggccagc ttccaggaag aaaagagcaa gctggatgct gaagtgtcca aatgggacga 3061 cagtggcaat gacatcattg tgctggccaa gcagatgtgc atgattatga tggagatgac 3121 agactttacc cgaggtaaag gaccactcaa aaatacatcg gatgtcatca gtgctgccaa 3181 gaaaattgct gaggcaggat ccaggatgga caagcttggc cgcaccattg cagaccattg 3241 ccccgactcg gcttgcaagc aggacctgct ggcctacctg caacgcatcg ccctctactg 3301 ccaccagctg aacatctgca gcaaggtcaa ggccgaggtg cagaatctcg gcggggagct 3361 tgttgtctct ggggtggaca gcgccatgtc cctgatccag gcagccaaga acttgatgaa 3421 tgctgtggtg cagacagtga aggcatccta cgtcgcctct accaaatacc aaaagtcaca 3481 gggtatggct tccctcaacc ttcctgctgt gtcatggaag atgaaggcac cagagaaaaa 3541 gccattggtg aagagagaga aacaggatga gacacagacc aagattaaac gggcatctca 3601 gaagaagcac gtgaacccgg tgcaggccct cagcgagttc aaagctatgg acagcatcct 3661 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga 3721 cgatgacaag tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag 3781 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 3841 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 3901 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 3961 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg 4021 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 4081 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 4141 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 4201 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 4261 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 4321 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 4381 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 4441 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga 4501 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 4561 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 4621 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 4681 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 4741 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 4801 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga 4861 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 4921 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 4981 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 5041 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 5101 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 5161 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 5221 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 5281 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 5341 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 5401 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 5461 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 5521 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 5581 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 5641 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 5701 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 5761 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 5821 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa 5881 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 5941 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 6001 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 6061 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 6121 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 6181 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 6241 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 6301 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 6361 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 6421 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 6481 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 6541 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 6601 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 6661 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 6721 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 6781 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 6841 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 6901 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg 6961 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 7021 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 7081 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 7141 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 7201 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 7261 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 7321 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 7381 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 7441 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 7501 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 7561 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 7621 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 7681 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 7741 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 7801 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 7861 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 7921 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 7981 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 8041 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 8101 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 8161 ctgacgtc //