LOCUS V022440 7109 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022440 VERSION V022440 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 683..1789 /label="ST13(NM_003932)" /note="ST13(NM_003932)" /gene="ST13" CDS 1796..1861 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1885..2473 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2539..2587 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2789..3217 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3231..3560 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3627..4421 /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 4597..4718 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4767..4783) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4791..4807 /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(4815..4845) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4860..4881 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5169..5757) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5928..6788) /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(6789..6893) /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 ggatccgcca ccatggaccc ccgcaaagtg aacgagcttc gggcctttgt 721 gaaaatgtgt aagcaggatc cgagcgttct gcacaccgag gaaatgcgct tcctgaggga 781 gtgggtggag agcatgggtg gtaaagtacc acctgctact cagaaagcta aatcagaaga 841 aaataccaag gaagaaaaac ctgatagtaa gaaggtggag gaagacttaa aggcagacga 901 accatcaagt gaggaaagtg atctagaaat tgataaagaa ggtgtgattg aaccagacac 961 tgatgctcct caagaaatgg gagatgaaaa tgcggagata acggaggaga tgatggatca 1021 ggcaaatgat aaaaaagtgg ctgctattga agccctaaat gatggtgaac tccagaaagc 1081 cattgactta ttcacagatg ccatcaagct gaatcctcgc ttggccattt tgtatgccaa 1141 gagggccagt gtcttcgtca aattacagaa gccaaatgct gccatccgag actgtgacag 1201 agccattgaa ataaatcctg attcagctca gccttacaag tggcggggga aagcacacag 1261 acttctaggc cactgggaag aagcagccca tgatcttgcc cttgcctgta aattggatta 1321 tgatgaagat gctagtgcaa tgctgaaaga agttcaacct agggcacaga aaattgcaga 1381 acatcggaga aagtatgagc gaaaacgtga agagcgagag atcaaagaaa gaatagaacg 1441 agttaagaag gctcgagaag agcatgagag agcccagagg gaggaagaag ccagacgaca 1501 gtcaggagct cagtatggct cttttccagg tggctttcct gggggaatgc ctggtaattt 1561 tcccggagga atgcctggaa tgggaggggg catgcctgga atggctggaa tgcctggact 1621 caatgaaatt cttagtgatc cagaggttct tgcagccatg caggatccag aagttatggt 1681 ggctttccag gatgtggctc agaacccagc aaatatgtca aaataccaga gcaacccaaa 1741 ggttatgaat ctcatcagta aattgtcagc caaatttgga ggtcaagcgc tcgaggacta 1801 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa 1861 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg 1921 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 1981 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 2041 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 2101 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 2161 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 2221 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag 2281 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 2341 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 2401 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 2461 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg 2521 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg 2581 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga 2641 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg 2701 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg 2761 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 2821 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 2881 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 2941 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 3001 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 3061 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 3121 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta 3181 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 3241 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3301 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3361 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 3421 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3481 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 3541 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 3601 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 3661 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 3721 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 3781 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 3841 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 3901 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 3961 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 4021 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 4081 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 4141 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 4201 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 4261 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 4321 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 4381 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc 4441 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 4501 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 4561 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat 4621 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 4681 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc 4741 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 4801 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 4861 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 4921 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 4981 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 5041 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 5101 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 5161 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 5221 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 5281 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 5341 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc 5401 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 5461 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 5521 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 5581 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag 5641 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 5701 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 5761 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 5821 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 5881 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 5941 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 6001 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 6061 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 6121 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 6181 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 6241 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 6301 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 6361 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 6421 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 6481 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 6541 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 6601 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 6661 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 6721 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 6781 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 6841 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 6901 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga 6961 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg 7021 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga 7081 ccgacaattg catgaagaat ctgcttagg //