LOCUS V021419 7649 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021419 VERSION V021419 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 647..2329 /label="Ces2c(NM_145603)" /note="Ces2c(NM_145603)" /gene="Ces2c" CDS 2336..2401 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2425..3013 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3079..3127 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3329..3757 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3771..4100 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4167..4961 /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 5137..5258 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5307..5323) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5331..5347 /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(5355..5385) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5400..5421 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5709..6297) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6468..7328) /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(7329..7433) /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 gccaccatga cacggaacca 661 actacataac tggctgaatg ctgggttctt cgggctgctg cttcttctta tccatgtgca 721 gggtcaggac tcaccagagg ccaaccccat cagaaacaca catacaggac agatccaagg 781 cagccttatc cacgtgaagg acactaaagc tggtgtccac accttcctgg gaatcccctt 841 tgccaaacct cctgtaggac cactgcgatt tgcacctcct gaggcccctg aaccatggag 901 tggtgtgaga gatggaactg cacatccggc catgtgtctg caaaatcttg acatgctgaa 961 tgaagcaggc ctgccagata tgaaaatgat gctgtcttcc ttccctatgt ctgaggactg 1021 cctgtatctc aacatctaca ccccagccca tgcccatgag ggctctaacc tgcctgtgat 1081 ggtgtggatc catggaggtg cactggttat aggaatggct tccatgtttg atggatctct 1141 attgacagtc aatgaggact tggtggttgt tactatccag tatcgtctgg gtgtcctggg 1201 ctttttcagc actggagacc agcatgccag aggcaactgg ggatacctgg accaagcagc 1261 tgccctacgc tgggtccagc agaacatcgc tcactttgga ggcaaccctg accgggtcac 1321 tatttttggc gagtctgcag gtggcacaag tgtgtcttca catgttgtgt cccccatgtc 1381 ccaagggctc ttccatggtg ccatcatgga gagtggggtg gccctgcttc ctgaccttat 1441 ctctgaaacc tctgagatgg tttccactac ggtggccaag ctttctggat gtgaggctat 1501 ggattcccag gccctggtgc gctgtctgag aggcaaaagt gaagcagaga ttctggctat 1561 taacaaggtc ttcaagatga tccctgctgt ggtggatggg gagttcttcc ccaggcatcc 1621 caaagagttg ttggcatctg aggattttca ccctgtcccc agcatcattg gtgtcaacaa 1681 tgatgagttt ggctggtcca ttcctgtggt catgggctct gctcagatga taaaaggaat 1741 aaccagagag aacctgcagg ctgttctgaa ggatacagca gtacaaatga tgctgcctcc 1801 tgagtgtagt gacctgctaa tggaagagta catgggggac actgaggatg cccagaccct 1861 ccaaatacag ttcacagaga tgatgggaga tttcatgttt gtgatccctg cactccaagt 1921 agcacatttt caacgctccc atgcccctgt ctacttctat gagttccaac atccacccag 1981 ctacttcaag gatgtcagac caccccacgt gaaggctgac catgctgatg agattccttt 2041 tgtctttgcg tccttcttct ggggcatgaa acttgacttc actgaggagg aggagctgct 2101 gagcaggagg atgatgaagt actgggccaa ctttgcacga catgggaacc ccaacagtga 2161 gggtctaccc tactggcctg tgatggacca tgatgagcag tacctgcaac tggacatcca 2221 gcctgctgtg ggtcgagccc tgaaggctgg aaggctgcag ttctggacca agactctgcc 2281 ccagaagatc caggagctaa aggcttctca ggacaagcac agggagcttc tcgaggacta 2341 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa 2401 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg 2461 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 2521 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 2581 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 2641 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 2701 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 2761 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag 2821 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 2881 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 2941 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 3001 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg 3061 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg 3121 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga 3181 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg 3241 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg 3301 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 3361 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 3421 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 3481 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 3541 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 3601 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 3661 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta 3721 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 3781 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3841 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3901 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 3961 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 4021 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 4081 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 4141 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 4201 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 4261 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 4321 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 4381 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 4441 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 4501 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 4561 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 4621 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 4681 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 4741 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 4801 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 4861 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 4921 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc 4981 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 5041 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 5101 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat 5161 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 5221 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc 5281 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 5341 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 5401 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 5461 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 5521 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 5581 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 5641 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 5701 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 5761 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 5821 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 5881 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc 5941 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 6001 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 6061 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 6121 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag 6181 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 6241 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 6301 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 6361 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 6421 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 6481 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 6541 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 6601 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 6661 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 6721 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 6781 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 6841 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 6901 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 6961 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 7021 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 7081 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 7141 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 7201 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 7261 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 7321 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 7381 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 7441 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga 7501 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg 7561 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga 7621 ccgacaattg catgaagaat ctgcttagg //