LOCUS V046899 5899 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046899 VERSION V046899 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 925..1650 /label="NAA60(NM_024845)" /note="NAA60(NM_024845)" /gene="NAA60" CDS 1663..1728 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1763..1987 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2033..2461 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2475..2804 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2816..3415 /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 3575..3696 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3745..3761) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3769..3785 /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(3793..3823) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3838..3859 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4147..4732) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4903..5763) /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(5764..5868) /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 aagctttcta gagaattcgc caccatgaca gaggtggtgc catccagcgc gctcagcgag 961 gtcagcctgc gcctcctctg ccacgatgac atagacactg tgaagcacct gtgtggcgac 1021 tggttcccca tcgagtaccc agactcatgg tatcgtgata tcacatccaa caagaagttc 1081 ttttcccttg ctgcaaccta cagaggtgcc attgtgggaa tgatagtagc tgaaattaag 1141 aacaggacca aaatacataa agaggatgga gatattctag catccaactt ctctgttgac 1201 acacaagtcg cgtacatcct aagtctgggc gtcgtgaaag agttcaggaa gcacggcata 1261 ggttccctct tacttgaaag tttaaaggat cacatatcaa ccaccgccca ggaccactgc 1321 aaagccattt acctgcatgt cctcaccacc aacaacacag caataaactt ctatgaaaac 1381 agagacttca agcagcacca ctatctcccc tattactact ccattcgagg ggtcctcaaa 1441 gatggcttca cctatgtcct ctacatcaac ggcggccacc ctccctggac gattttggac 1501 tacatccagc acctgggctc tgcactagcc agcctgagcc cctgctccat tccgcacaga 1561 gtctaccgcc aggcccacag cctgctctgc agcttcctgc catggtcggg catctcttcc 1621 aagagtggca tcgagtacag ccggaccatg ctcgagaccg gtgactacaa agaccatgac 1681 ggtgattata aagatcatga catcgattac aaggatgacg atgacaagtg accggtaagt 1741 ttaaaccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc 1801 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat 1861 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg 1921 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc 1981 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc 2041 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 2101 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 2161 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 2221 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 2281 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 2341 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 2401 ttggccattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 2461 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 2521 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 2581 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 2641 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 2701 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc 2761 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggatgac 2821 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac 2881 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca 2941 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 3001 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 3061 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 3121 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 3181 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 3241 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 3301 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 3361 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgattcga 3421 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt 3481 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg 3541 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg 3601 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 3661 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc 3721 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 3781 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 3841 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 3901 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 3961 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 4021 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 4081 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 4141 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 4201 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 4261 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 4321 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 4381 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 4441 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 4501 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 4561 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 4621 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 4681 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 4741 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 4801 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 4861 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 4921 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 4981 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 5041 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 5101 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 5161 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 5221 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 5281 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 5341 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 5401 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 5461 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 5521 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 5581 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 5641 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 5701 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 5761 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 5821 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 5881 aaaagtgcca cctgacgtc //