LOCUS V040888 7590 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040888 VERSION V040888 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 141..157 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 318..697 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 698..901 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 931..996 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 1000..2295 /label="AHCY(NM_000687)" /note="AHCY(NM_000687)" /gene="AHCY" polyA_signal 2349..2971 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" promoter 3013..3310 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3364..4158 /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 4810..4944 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" promoter complement(4950..4968) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(4982..4998) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5006..5022 /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(5030..5060) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5075..5096 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5384..5972) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6143..7003) /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(7004..7108) /gene="bla" /label="AmpR promoter" rep_origin 7135..7563 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa 181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta 241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg 361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa 961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tgtctgacaa actgccctac 1021 aaagtcgccg acatcggcct ggctgcctgg ggacgcaagg ccctggacat tgctgagaac 1081 gagatgccgg gcctgatgcg tatgcgggag cggtactcgg cctccaagcc actgaagggc 1141 gcccgcatcg ctggctgcct gcacatgacc gtggagacgg ccgtcctcat tgagaccctc 1201 gtcaccctgg gtgctgaggt gcagtggtcc agctgcaaca tcttctccac ccaggaccat 1261 gcggcggctg ccattgccaa ggctggcatt ccggtgtatg cctggaaggg cgaaacggac 1321 gaggagtacc tgtggtgcat tgagcagacc ctgtacttca aggacgggcc cctcaacatg 1381 attctggacg acgggggcga cctcaccaac ctcatccaca ccaagtaccc gcagcttctg 1441 ccaggcatcc gaggcatctc tgaggagacc acgactgggg tccacaacct ctacaagatg 1501 atggccaatg ggatcctcaa ggtgcctgcc atcaatgtca atgactccgt caccaagagc 1561 aagtttgaca acctctatgg ctgccgggag tccctcatag atggcatcaa gcgggccaca 1621 gatgtgatga ttgccggcaa ggtagcggtg gtagcaggct atggtgatgt gggcaagggc 1681 tgtgcccagg ccctgcgggg tttcggagcc cgcgtcatca tcaccgagat tgaccccatc 1741 aacgcactgc aggctgccat ggagggctat gaggtgacca ccatggatga ggcctgtcag 1801 gagggcaaca tctttgtcac caccacaggc tgtattgaca tcatccttgg ccggcacttt 1861 gagcagatga aggatgatgc cattgtgtgt aacattggac actttgacgt ggagatcgat 1921 gtcaagtggc tcaacgagaa cgccgtggag aaggtgaaca tcaagccgca ggtggaccgg 1981 tatcggttga agaatgggcg ccgcatcatc ctgctggccg agggtcggct ggtcaacctg 2041 ggttgtgcca tgggccaccc cagcttcgtg atgagtaact ccttcaccaa ccaggtgatg 2101 gcgcagatcg agctgtggac ccatccagac aagtaccccg ttggggttca tttcctgccc 2161 aagaagctgg atgaggcagt ggctgaagcc cacctgggca agctgaatgt gaagttgacc 2221 aagctaactg agaagcaagc ccagtacctg ggcatgtcct gtgatggccc cttcaagccg 2281 gatcactacc gctactgaga attcatcgat agatctgata tcggtaccag tcgactctag 2341 aggatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg 2401 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact 2461 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg 2521 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac 2581 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc 2641 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt 2701 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct 2761 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg 2821 tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac 2881 ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg 2941 ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg 3001 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3061 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 3121 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 3181 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 3241 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 3301 gcttttttgg aggaattgat cagcttggga tctgatcaag agacaggatg aggatcgttt 3361 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 3421 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 3481 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 3541 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 3601 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 3661 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 3721 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 3781 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 3841 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 3901 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 3961 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 4021 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 4081 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4141 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 4201 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 4261 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 4321 tcgcccaccc cgggctcgat cccctcgcga gttggttcag ctgctgcctg aggctggacg 4381 acctcgcgga gttctaccgg cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct 4441 atccgcgcat ccatgccccc gaactgcagg agtggggagg cacgatggcc gctttggtcg 4501 acccggacgg gacgctcctg cgcctgatac agaacgaatt gcttgcaggc atctcatgag 4561 tgtgtcttcc cgttttccgc ctgaggtcac tgcgtggatg gagcgctggc gcctgctgcg 4621 cgacggcgag ctgctcacca cccactcgcc aagctggaac cgtaaaaagg ccgcgttgct 4681 ggcgtttttc cataggctcc gccgatcata atcagccata ccacatttgt agaggtttta 4741 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt 4801 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 4861 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 4921 aatgtatctt atcatgtctg gatcaattcc ctatagtgag tcgtattaaa ttcgtaatca 4981 tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 5041 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 5101 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 5161 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 5221 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 5281 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 5341 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 5401 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 5461 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 5521 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 5581 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 5641 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 5701 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 5761 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 5821 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 5881 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 5941 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 6001 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 6061 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 6121 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 6181 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 6241 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 6301 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 6361 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 6421 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 6481 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 6541 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 6601 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 6661 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 6721 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 6781 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 6841 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 6901 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 6961 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 7021 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 7081 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc 7141 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 7201 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 7261 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 7321 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 7381 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 7441 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 7501 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 7561 attttaacaa aatattaacg cttacaattt //