LOCUS V029767 7142 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029767 VERSION V029767 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 908..2584 /label="CES2(NM_001365405)" /note="CES2(NM_001365405)" /gene="CES2" CDS 2585..2650 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2709..2933 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2979..3407 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3421..3750 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3817..4611 /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 4785..4906 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4955..4971) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4979..4995 /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(5003..5033) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5048..5069 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5357..5942) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6113..6973) /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(6974..7078) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cgccaccatg cggctgcaca gacttcgtgc gcggctgagc gcggtggcct gtgggcttct 961 gctgcttctt gtccggggcc agggccagga ctcagccagt cccatccgga ccacacacac 1021 ggggcaggtg ctggggagtc ttgtccatgt gaagggcgcc aatgccgggg tccaaacctt 1081 cctgggaatt ccatttgcca agccacctct aggtccgctg cgatttgcac cccctgagcc 1141 ccctgaatct tggagtggtg tgagggatgg aaccacccat ccggccatgt gtctacagga 1201 cctcaccgca gtggagtcag agtttcttag ccagttcaac atgaccttcc cttccgactc 1261 catgtctgag gactgcctgt acctcagcat ctacacgccg gcccatagcc atgaaggctc 1321 taacctgccg gtgatggtgt ggatccacgg tggtgcgctt gtttttggca tggcttcctt 1381 gtatgatggt tccatgctgg ctgccttgga gaacgtggtg gtggtcatca tccagtaccg 1441 cctgggtgtc ctgggcttct tcagcactgg agacaagcac gcaaccggca actggggcta 1501 cctggaccaa gtggctgcac tacgctgggt ccagcagaat atcgcccact ttggaggcaa 1561 ccctgaccgt gtcaccattt ttggcgagtc tgcgggtggc acgagtgtgt cttcgcttgt 1621 tgtgtccccc atatcccaag gactcttcca cggagccatc atggagagtg gcgtggccct 1681 cctgcccggc ctcattgcca gctcagctga tgtcatctcc acggtggtgg ccaacctgtc 1741 tgcctgtgac caagttgact ctgaggccct ggtgggctgc ctgcggggca agagtaaaga 1801 ggagattctt gcaattaaca agcctttcaa gatgatcccc ggagtggtgg atggggtctt 1861 cctgcccagg cacccccagg agctgctggc ctctgccgac tttcagcctg tccctagcat 1921 tgttggtgtc aacaacaatg aattcggctg gctcatcccc aaggtcatga ggatctatga 1981 tacccagaag gaaatggaca gagaggcctc ccaggctgct ctgcagaaaa tgttaacgct 2041 gctgatgttg cctcctacat ttggtgacct gctgagggag gagtacattg gggacaatgg 2101 ggatccccag accctccaag cgcagttcca ggagatgatg gcggactcca tgtttgtgat 2161 ccctgcactc caagtagcac attttcagtg ttcccgggcc cctgtgtact tctacgagtt 2221 ccagcatcag cccagctggc tcaagaacat caggccaccg cacatgaagg cagaccatgg 2281 tgatgagctt ccttttgttt tcagaagttt ctttgggggc aactacatta aattcactga 2341 ggaagaggag cagctaagca ggaagatgat gaagtactgg gccaactttg cgagaaatgg 2401 gaaccccaat ggcgagggtc tgccacactg gccgctgttc gaccaggagg agcaatacct 2461 gcagctgaac ctacagcctg cggtgggccg ggctctgaag gcccacaggc tccagttctg 2521 gaagaaggcg ctgccccaaa agatccagga gctcgaggag cctgaagaga gacacacaga 2581 gctggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga 2641 cgatgacaag aaggacgagc tgtagctcga gtctagaggg cccgtttaaa cccgctgatc 2701 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc 2761 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc 2821 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg 2881 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga 2941 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 3001 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 3061 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 3121 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 3181 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 3241 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 3301 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 3361 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 3421 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3481 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 3541 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 3601 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 3661 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 3721 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc 3781 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 3841 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 3901 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 3961 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 4021 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 4081 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 4141 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 4201 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 4261 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 4321 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 4381 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 4441 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 4501 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 4561 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 4621 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 4681 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 4741 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag 4801 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 4861 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac 4921 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 4981 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 5041 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 5101 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 5161 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 5221 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 5281 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 5341 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 5401 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 5461 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 5521 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 5581 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 5641 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 5701 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 5761 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 5821 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 5881 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5941 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 6001 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 6061 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 6121 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 6181 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 6241 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 6301 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 6361 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 6421 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 6481 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 6541 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 6601 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 6661 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 6721 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 6781 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 6841 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 6901 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 6961 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 7021 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 7081 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg 7141 gt //