LOCUS V027007 8866 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027007 VERSION V027007 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 706..3525 /label="AP2A2(NM_001242837)" /note="AP2A2(NM_001242837)" /gene="AP2A2" CDS 3532..3558 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3562..3588 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3592..3618 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" misc_feature 3642..4230 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4296..4344 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4546..4974 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4988..5317 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5384..6178 /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 6354..6475 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6524..6540) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6548..6564 /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(6572..6602) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6617..6638 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6926..7514) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7685..8545) /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(8546..8650) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgcc ggccgtgtcc 721 aagggggacg ggatgcgggg cctggcggtc ttcatctcgg atatccgcaa ctgtaaaagt 781 aaagaagcag aaataaaaag gataaacaag gaactggcaa atatcagatc aaaatttaaa 841 ggtgacaagg ctcttgatgg ctatagtaaa aaaaagtacg tctgcaagtt gctcttcatc 901 tttctccttg gtcatgacat tgactttgga cacatggagg ctgtgaacct gctgagttca 961 aacagataca cggaaaagca gatcggctac cttttcatct ctgtgttggt gaactcaaac 1021 agtgagctga tccgcctgat caacaacgcc atcaagaatg acctggccag ccgcaacccc 1081 accttcatgg gcctggccct gcactgcatc gccagcgtgg gcagccggga gatggccgag 1141 gccttcgccg gggagatccc taaggtcctc gtagccggag acactatgga cagcgtgaag 1201 cagagcgcgg ccctgtgctt gctgcgcctg tacaggacgt cccccgatct tgtccccatg 1261 ggcgactgga catcccgagt ggtgcacctg ctcaatgacc agcacttggg tgtggtaact 1321 gcagccacaa gtctgatcac cactttagca cagaagaacc cagaagagtt taaaacctcc 1381 gtgtctctgg ctgtctctag gctaagcaga atcgtgacgt ctgcatccac agatctccag 1441 gattacactt actattttgt cccggctccc tggctgtctg tcaaactgct gagactgctg 1501 cagtgctacc cacccccaga agaccctgca gtgcgaggcc gcctgactga gtgcctggag 1561 accatcctga acaaagccca agaaccgccc aagtcgaaga aggtccagca ctccaacgcg 1621 aagaatgccg tgctcttcga ggccatcagc ttaatcattc accatgacag tgagccgaac 1681 ctgctcgtcc gtgcctgcaa ccagttgggc cagtttctgc agcaccgcga gaccaacctg 1741 cgctacctgg ccctggagag catgtgcacg ctggccagct ctgagttctc ccatgaggct 1801 gtcaagacgc acatcgagac ggtcatcaac gccctgaaga ctgagcggga cgtgagcgtg 1861 cggcagcggg ccgtggacct cctctacgcc atgtgcgacc gcagcaacgc cccacagatc 1921 gtggccgaga tgctgagcta tctggagaca gctgactact ccatccgaga agagattgtg 1981 ctgaaggtcg ccatcctggc tgagaagtac gcggtggact acacctggta tgtggatacc 2041 atcttgaact tgatccgaat tgctggtgat tacgtgagtg aagaggtgtg gtaccgagtc 2101 attcagatcg tcatcaaccg ggacgacgtg cagggctacg cggccaagac tgtgttcgag 2161 gctcttcagg ctcccgcgtg ccacgagaac ctggtcaaag tgggcggcta catcctgggg 2221 gagtttggaa acttgatagc tggagacccg agatccagcc cgctgatcca gttccacctg 2281 ctgcactcca agttccacct gtgcagcgtc cccacccgcg cgctgctcct gtccacctac 2341 atcaagttcg tgaacctctt cccggaggtg aagcccacca tccaggacgt gctgcgcagc 2401 gacagccagc tcaggaacgc agacgtggag ctgcagcagc gtgctgtgga gtacctgcgg 2461 ctcagcaccg tggccagcac cgacattctg gcgaccgtgc tggaggagat gcccccattc 2521 ccggagcggg agtcctccat cttggcaaag ctcaagaaga agaagggccc cagcacggtg 2581 acagacctgg aggacaccaa gcgggacagg agtgtggacg tgaacggggg tcctgagcct 2641 gccccagcca gtaccagcgc cgtgtctacg ccttctccgt cggcagacct gctgggtctc 2701 ggggctgccc cccctgcccc cgcgggcccc ccaccctcct ccggcggcag cgggctgctc 2761 gtggacgtgt tctcagactc ggcctctgtg gtcgcgcctc tcgctcctgg ctccgaagac 2821 aactttgcca ggtttgtttg taaaaacaat ggtgtgttgt ttgaaaacca gctgcttcaa 2881 attggactta agtctgaatt tcggcagaat ttaggtcgga tgtttatctt ttatggtaat 2941 aagacctcca cgcagttcct aaactttacc ccaacactaa tctgttcaga cgaccttcag 3001 cctaacctga acctgcagac caagcccgtg gacccgaccg tggagggggg cgcgcaggtg 3061 cagcaggtgg tcaacataga gtgcgtgtcc gacttcacgg aggcgccagt cctcaacatt 3121 cagttcaggt atgggggcac cttccagaac gtgtctgtgc agctgcccat cactctcaac 3181 aaattcttcc agccgacaga aatggcttct caggatttct ttcaacgttg gaagcagttg 3241 agcaatccac agcaggaagt gcagaacatc ttcaaagcaa agcacccaat ggacacagaa 3301 gtcaccaaag ccaagatcat tggatttggt tctgcacttc ttgaagaagt tgatcctaat 3361 cctgcgaatt tcgtgggagc tggaatcatc cacacgaaaa ccacccagat tggatgcctg 3421 ctgcgcttgg agccgaacct gcaagcccag atgtaccggc tcacgctgcg cacaagtaag 3481 gaagccgttt ctcagagatt atgtgaattg ctctcagcgc agtttctcga gtacccatac 3541 gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag ctatccttat 3601 gatgtcccag actacgcata gtgagggccc gatatctcga caatcaacct ctggattaca 3661 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 3721 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 3781 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 3841 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 3901 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca 3961 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 4021 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga 4081 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 4141 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga 4201 gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg 4261 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac 4321 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg 4381 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac 4441 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc 4501 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg 4561 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 4621 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 4681 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc 4741 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga 4801 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 4861 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga 4921 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct 4981 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 5041 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 5101 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac 5161 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 5221 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta 5281 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc 5341 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg 5401 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca 5461 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt 5521 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg 5581 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga 5641 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca 5701 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct 5761 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac 5821 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc 5881 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt 5941 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt 6001 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg 6061 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat 6121 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc 6181 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 6241 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 6301 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt 6361 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 6421 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 6481 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 6541 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 6601 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 6661 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 6721 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 6781 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 6841 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 6901 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 6961 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 7021 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 7081 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc 7141 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 7201 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 7261 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 7321 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta 7381 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 7441 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 7501 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 7561 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 7621 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 7681 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 7741 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 7801 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 7861 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 7921 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 7981 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 8041 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 8101 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 8161 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 8221 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 8281 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 8341 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 8401 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 8461 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 8521 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 8581 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 8641 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc 8701 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta 8761 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac 8821 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg //