LOCUS V036257 6709 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036257 VERSION V036257 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..1368 /label="H1-3(NM_005320)" /note="H1-3(NM_005320)" /gene="H1-3" CDS 1375..1401 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 1405..1431 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 1435..1461 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" misc_feature 1485..2073 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2139..2187 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2389..2817 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2831..3160 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3227..4021 /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 4197..4318 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4367..4383) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4391..4407 /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(4415..4445) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4460..4481 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4769..5357) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5528..6388) /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(6389..6493) /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 ggagactgct 721 ccacttgctc ctaccattcc tgcacccgca gaaaaaacac ctgtgaagaa aaaggcgaag 781 aaggcaggcg caactgctgg gaaacgcaaa gcatccggac ccccagtatc tgagcttatc 841 accaaggcag tggcagcttc taaggagcgc agcggcgttt ctctggccgc gcttaagaaa 901 gcgcttgcgg ctgctggcta cgatgtagaa aaaaacaaca gccgtatcaa gcttggcctc 961 aagagcttgg tgagcaaagg tactctggtg cagaccaaag gtaccggtgc ttctggctcc 1021 ttcaaactca acaagaaagc ggcttccggg gaaggcaaac ccaaggccaa aaaggctggc 1081 gcagccaagc ctaggaagcc tgctggggca gccaagaagc ccaagaaggt ggctggcgcc 1141 gctaccccga agaaaagcat caaaaagact cctaagaagg taaagaagcc agcaaccgct 1201 gctgggacca agaaagtggc caagagtgcg aaaaaggtga aaacacctca gccaaaaaaa 1261 gctgccaaga gtccagctaa ggccaaagcc cctaagccca aggcggccaa gcctaagtcg 1321 gggaagccga aggttacaaa ggcaaagaag gcagctccga agaaaaagct cgagtaccca 1381 tacgacgtac ctgattatgc aacttatcct tatgatgtac ctgattatgc tagctatcct 1441 tatgatgtcc cagactacgc atagtgaggg cccgatatct cgacaatcaa cctctggatt 1501 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 1561 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 1621 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 1681 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 1741 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 1801 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 1861 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct 1921 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 1981 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 2041 cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac 2101 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa 2161 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact 2221 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc 2281 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag 2341 gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta 2401 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca 2461 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct 2521 ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc 2581 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat 2641 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc 2701 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg 2761 ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat 2821 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag 2881 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 2941 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 3001 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 3061 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa 3121 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat 3181 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 3241 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 3301 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 3361 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc 3421 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 3481 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 3541 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 3601 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 3661 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 3721 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt 3781 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 3841 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 3901 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 3961 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 4021 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag 4081 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 4141 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact 4201 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 4261 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 4321 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc 4381 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 4441 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 4501 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg 4561 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 4621 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 4681 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 4741 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 4801 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 4861 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 4921 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt 4981 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 5041 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 5101 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 5161 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg 5221 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 5281 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 5341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 5401 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 5461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 5521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 5581 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 5641 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 5701 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 5761 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 5821 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 5881 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 5941 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 6001 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 6061 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 6121 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 6181 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 6241 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 6301 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 6361 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 6421 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 6481 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc 6541 tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca 6601 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc 6661 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg //