LOCUS V019749 7616 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019749 VERSION V019749 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 683..2296 /label="NECTIN2(NM_001042724)" /note="NECTIN2(NM_001042724)" /gene="NECTIN2" CDS 2303..2368 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2392..2980 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3046..3094 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3296..3724 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3738..4067 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4134..4928 /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 5104..5225 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5274..5290) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5298..5314 /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(5322..5352) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5367..5388 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5676..6264) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6435..7295) /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(7296..7400) /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 ggatccgcca ccatggcccg ggccgctgcc ctcctgccgt cgagatcgcc 721 gccgacgccg ctgctgtggc cgctgctgct gctgctgctc ctggaaaccg gagcccagga 781 tgtgcgagtt caagtgctac ccgaggtgcg aggccagctc gggggcaccg tggagctgcc 841 gtgccacctg ctgccacctg ttcctggact gtacatctcc ctggtgacct ggcagcgccc 901 agatgcacct gcgaaccacc agaatgtggc cgccttccac cctaagatgg gtcccagctt 961 ccccagcccg aagcctggca gcgagcggct gtccttcgtc tctgccaagc agagcactgg 1021 gcaagacaca gaggcagagc tccaggacgc cacgctggcc ctccacgggc tcacggtgga 1081 ggacgagggc aactacactt gcgagtttgc caccttcccc aaggggtccg tccgagggat 1141 gacctggctc agagtcatag ccaagcccaa gaaccaagct gaggcccaga aggtcacgtt 1201 cagccaggac cctacgacag tggccctctg catctccaaa gagggccgcc cacctgcccg 1261 gatctcctgg ctctcatccc tggactggga agccaaagag actcaggtgt cagggaccct 1321 ggccggaact gtcactgtca ccagccgctt caccttggtg ccctcgggcc gagcagatgg 1381 tgtcacggtc acctgcaaag tggagcatga gagcttcgag gaaccagccc tgatacctgt 1441 gaccctctct gtacgctacc ctcctgaagt gtccatctcc ggctatgatg acaactggta 1501 cctcggccgt actgatgcca ccctgagctg tgacgtccgc agcaacccag agcccacggg 1561 ctatgactgg agcacgacct caggcacctt cccgacctcc gcagtggccc agggctccca 1621 gctggtcatc cacgcagtgg acagtctgtt caataccacc ttcgtctgca cagtcaccaa 1681 tgccgtgggc atgggccgcg ctgagcaggt catctttgtc cgagagaccc ccaacacagc 1741 aggcgcaggg gccacaggcg gcatcatcgg gggcatcatc gccgccatca ttgctactgc 1801 tgtggctgcc acgggcatcc ttatctgccg gcagcagcgg aaggagcaga cgctgcaggg 1861 ggcagaggag gacgaagacc tggagggacc tccctcctac aagccaccga ccccaaaagc 1921 gaagctggag gcacaggaga tgccctccca gctcttcact ctgggggcct cggagcacag 1981 cccactcaag accccctact ttgatgctgg cgcctcatgc actgagcagg aaatgcctcg 2041 ataccatgag ctgcccacct tggaagaacg gtcaggaccc ttgcaccctg gagccacaag 2101 cctggggtcc cccatcccgg tgcctccagg gccacctgct gtggaagacg tttccctgga 2161 tctagaggat gaggaggggg aggaggagga agagtatctg gacaagatca accccatcta 2221 tgatgctctg tcctatagca gcccctctga ttcctaccag ggcaaaggct ttgtcatgtc 2281 ccgggccatg tatgtgctcg aggactacaa agaccatgac ggtgattata aagatcatga 2341 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct 2401 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 2461 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 2521 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 2581 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 2641 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 2701 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 2761 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt 2821 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 2881 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 2941 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa 3001 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac 3061 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc 3121 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc 3181 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg 3241 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg 3301 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 3361 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 3421 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct 3481 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 3541 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 3601 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 3661 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 3721 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 3781 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 3841 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 3901 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3961 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 4021 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 4081 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 4141 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 4201 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 4261 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 4321 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 4381 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 4441 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 4501 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 4561 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 4621 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 4681 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 4741 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 4801 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 4861 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 4921 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca 4981 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 5041 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 5101 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 5161 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 5221 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 5281 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 5341 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 5401 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 5461 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 5521 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 5581 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 5641 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 5701 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 5761 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 5821 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc 5881 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 5941 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 6001 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 6061 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 6121 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 6181 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 6241 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 6301 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 6361 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 6421 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 6481 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 6541 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 6601 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 6661 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 6721 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 6781 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 6841 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 6901 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 6961 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 7021 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 7081 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 7141 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 7201 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 7261 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 7321 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 7381 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg 7441 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt 7501 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa 7561 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg //