LOCUS V020850 6359 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020850 VERSION V020850 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 905..2110 /label="Sirt7(NM_153056)" /note="Sirt7(NM_153056)" /gene="Sirt7" CDS 2123..2188 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2223..2447 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2493..2921 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2935..3264 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3276..3875 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" polyA_signal 4035..4156 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4205..4221) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4229..4245 /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(4253..4283) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4298..4319 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4607..5192) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5363..6223) /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(6224..6328) /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 caccatggca gccggtggcg gtctgagccg ctcggagcgc aaagctgctg agcgggtccg 961 gaggctgcgg gaggagcagc agcgggagcg cctccgccag gtgtcacgca tcctgaggaa 1021 ggcggctgca gagcgcagcg cggaggaggg ccggctcttg gccgagagcg aggatctggt 1081 gaccgagctg cagggtcgaa gtcggcggcg tgagggcctc aagcgccgcc aggaggaggt 1141 gtgtgatgac ccggaggagc tgcggaggaa ggtccgcgaa ctggccggag ctgtccgaag 1201 tgccaggcac ttggttgtct acacgggcgc tggaatcagc acagcagctt ctatcccaga 1261 ttatcggggt cctaatggag tatggacact gcttcagaaa ggaaggcctg tgagtgctgc 1321 cgacctaagc gaagcggagc ctaccctcac ccacatgagc atcacccgtt tgcatgagca 1381 aaagctggtg caacacgtgg tgtctcagaa ctgtgatggg ctccacctgc ggagtgggtt 1441 gccccggacc gccatctcag agctccatgg gaatatgtat attgaagtct gcacctcctg 1501 catccctaac agagagtatg tacgagtgtt tgatgtgact gagcgtactg cccttcaccg 1561 acacctgaca ggccggacct gccacaagtg cgggacccag cttcgggata ccattgtgca 1621 ctttggggag agggggacat tagggcagcc tctgaactgg gaggcagcga ccgaggctgc 1681 tagcaaagca gacacaatcc tgtgtttagg gtccagcttg aaggtactaa agaaatatcc 1741 ccgcctctgg tgcatgacga agcctccaag ccgtcgaccc aaactctaca tcgtgaacct 1801 gcagtggacc ccgaaggatg actgggctgc cctgaaactc catgggaagt gtgatgatgt 1861 catgcaactc ctcatgaatg aactgggcct ggagattcct gtctacaacc ggtggcagga 1921 tccaatcttc tccttggcga cccccctccg tgctggtgaa gaaggcagcc acagtaggaa 1981 gtcactatgc agaagcagag aagaagcccc acctggggac cagagtgacc cccttgcctc 2041 agctccccct atcctaggag gctggtttgg caggggttgt gccaagcgtg caaaaaggaa 2101 gaaagtggca ctcgagaccg gtgactacaa agaccatgac ggtgattata aagatcatga 2161 catcgattac aaggatgacg atgacaagtg accggtaagt ttaaaccgct gatcagcctc 2221 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac 2281 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg 2341 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga 2401 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga 2461 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc 2521 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 2581 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 2641 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 2701 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 2761 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 2821 caaccctatc tcggtctatt cttttgattt ataagggatt ttggccattt cggcctattg 2881 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt 2941 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 3001 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg 3061 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg 3121 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 3181 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt 3241 tttggaggcc taggcttttg caaaaagctc ccgggatgac cgagtacaag cccacggtgc 3301 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg 3361 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc 3421 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg 3481 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg 3541 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga 3601 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg 3661 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg 3721 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc 3781 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc 3841 gcacctggtg catgacccgc aagcccggtg cctgattcga aatgaccgac caagcgacgc 3901 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg 3961 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt 4021 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 4081 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 4141 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat 4201 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 4261 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 4321 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 4381 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 4441 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 4501 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 4561 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 4621 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 4681 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 4741 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 4801 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 4861 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 4921 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 4981 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 5041 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 5101 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc 5161 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 5221 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 5281 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 5341 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 5401 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 5461 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 5521 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 5581 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 5641 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 5701 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 5761 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 5821 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 5881 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 5941 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 6001 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 6061 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 6121 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 6181 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 6241 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 6301 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc //