LOCUS V039665 7039 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039665 VERSION V039665 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 706..1695 /label="PPP1CA(NM_002708)" /note="PPP1CA(NM_002708)" /gene="PPP1CA" CDS 1702..1791 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 1815..2403 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2469..2517 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2719..3147 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3161..3490 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3557..4351 /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 4527..4648 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4697..4713) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4721..4737 /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(4745..4775) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4790..4811 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5099..5687) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5858..6718) /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(6719..6823) /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgtc cgacagcgag 721 aagctcaacc tggactcgat catcgggcgc ctgctggaag tgcagggctc gcggcctggc 781 aagaatgtac agctgacaga gaacgagatc cgcggtctgt gcctgaaatc ccgggagatt 841 tttctgagcc agcccattct tctggagctg gaggcacccc tcaagatctg cggtgacata 901 cacggccagt actacgacct tctgcgacta tttgagtatg gcggtttccc tcccgagagc 961 aactacctct ttctggggga ctatgtggac aggggcaagc agtccttgga gaccatctgc 1021 ctgctgctgg cctataagat caagtacccc gagaacttct tcctgctccg tgggaaccac 1081 gagtgtgcca gcatcaaccg catctatggt ttctacgatg agtgcaagag acgctacaac 1141 atcaaactgt ggaaaacctt cactgactgc ttcaactgcc tgcccatcgc ggccatagtg 1201 gacgaaaaga tcttctgctg ccacggaggc ctgtccccgg acctgcagtc tatggagcag 1261 attcggcgga tcatgcggcc cacagatgtg cctgaccagg gcctgctgtg tgacctgctg 1321 tggtctgacc ctgacaagga cgtgcagggc tggggcgaga acgaccgtgg cgtctctttt 1381 acctttggag ccgaggtggt ggccaagttc ctccacaagc acgacttgga cctcatctgc 1441 cgagcacacc aggtggtaga agacggctac gagttctttg ccaagcggca gctggtgaca 1501 cttttctcag ctcccaacta ctgtggcgag tttgacaatg ctggcgccat gatgagtgtg 1561 gacgagaccc tcatgtgctc tttccagatc ctcaagcccg ccgacaagaa caaggggaag 1621 tacgggcagt tcagtggcct gaaccctgga ggccgaccca tcaccccacc ccgcaattcc 1681 gccaaagcca agaaactcga ggaacaaaaa ctaatatcag aggaagattt ggaacagaaa 1741 ctaatcagtg aagaagattt agagcaaaag ttaatttccg aggaggactt atagtgaggg 1801 cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc 1861 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg 1921 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc 1981 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg 2041 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg 2101 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga 2161 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct 2221 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg 2281 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc 2341 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc 2401 cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg 2461 cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata 2521 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg 2581 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc 2641 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg 2701 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 2761 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 2821 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc 2881 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 2941 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 3001 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 3061 tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc 3121 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg 3181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 3241 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 3301 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 3361 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 3421 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 3481 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg 3541 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg 3601 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 3661 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 3721 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt 3781 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 3841 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 3901 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 3961 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 4021 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa 4081 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa 4141 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc 4201 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga 4261 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc 4321 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg 4381 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa 4441 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc 4501 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat 4561 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 4621 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga 4681 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 4741 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 4801 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 4861 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 4921 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 4981 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 5041 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 5101 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 5161 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 5221 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 5281 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 5341 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 5401 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 5461 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 5521 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 5581 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 5641 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 5701 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 5761 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 5821 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 5881 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 5941 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 6001 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 6061 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 6121 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 6181 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 6241 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 6301 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 6361 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 6421 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 6481 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 6541 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 6601 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 6661 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 6721 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 6781 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 6841 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac 6901 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt 6961 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg 7021 catgaagaat ctgcttagg //