LOCUS V032369 6991 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032369 VERSION V032369 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..1650 /label="Eif2s1(NM_019356)" /note="Eif2s1(NM_019356)" /gene="Eif2s1" CDS 1657..1683 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 1687..1713 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 1717..1743 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" misc_feature 1767..2355 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2421..2469 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2671..3099 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3113..3442 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3509..4303 /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 4479..4600 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4649..4665) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4673..4689 /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(4697..4727) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4742..4763 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5051..5639) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5810..6670) /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(6671..6775) /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 ccaccatgcc gggtctaagt 721 tgtagatttt atcaacacaa atttcctgag gtcgaagatg tagtgatggt gaatgtaaga 781 tccattgctg aaatgggggc ctatgtcagc ttgttggaat ataataacat tgaaggcatg 841 attcttctta gtgaattatc cagacgacgt atccgttcta taaacaaact gatccgaatt 901 ggcagaaatg aatgtgtagt tgtcattaga gtggataaag aaaaaggata tatagatttg 961 tcaaaaagaa gagtttctcc agaggaagca atcaaatgtg aagacaaatt cacaaaatcc 1021 aaaactgttt atagcattct tcgccatgtt gctgaggtat tagagtatac caaggatgag 1081 cagctggaaa gcctattcca gaggactgcc tgggtctttg atgacaagta caagagacct 1141 ggatatggtg cctatgatgc ctttaagcat gcagtctcag acccatctat cttggatagt 1201 ttagatttga atgaagatga aagagaagta ctcattaaca atatcaatag gcgtttgacc 1261 ccacaagctg tcaagattcg agcagatatt gaggtagctt gctatggtta cgaaggcatt 1321 gatgctgtaa aagaagccct gagagcaggt ttgaattgtt ctacagaaac catgcccatc 1381 aagattaatc taatagctcc acccaggtat gtgatgacaa caacgaccct agagaggaca 1441 gaaggactct ctgttctcaa tcaggctatg gcagtcatca aagaaaagat tgaggagaag 1501 aggggagtgt tcaatgttca gatggagccc aaagtggtta cagatacaga tgagactgaa 1561 cttgcaaggc agctggaacg gcttgagaga gaaaatgcag aagtggatgg agatgatgat 1621 gcagaagaaa tggaagccaa agctgaagat ctcgagtacc catacgacgt acctgattat 1681 gcaacttatc cttatgatgt acctgattat gctagctatc cttatgatgt cccagactac 1741 gcatagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat 1801 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc 1861 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct 1921 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca 1981 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt 2041 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg 2101 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga 2161 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt 2221 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc 2281 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt 2341 gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc 2401 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt 2461 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga 2521 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt 2581 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc 2641 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg 2701 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 2761 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 2821 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 2881 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 2941 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 3001 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt 3061 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca 3121 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 3181 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 3241 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 3301 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 3361 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt 3421 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga 3481 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc 3541 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 3601 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 3661 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc 3721 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 3781 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 3841 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 3901 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 3961 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 4021 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 4081 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 4141 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 4201 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 4261 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc 4321 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg 4381 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc 4441 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata 4501 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 4561 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga 4621 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 4681 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 4741 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 4801 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 4861 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 4921 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 4981 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 5041 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 5101 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 5161 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 5221 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg 5281 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 5341 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 5401 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 5461 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 5521 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 5581 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 5641 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 5701 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 5761 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 5821 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 5881 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 5941 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 6001 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 6061 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 6121 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 6181 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 6241 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 6301 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 6361 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 6421 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 6481 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 6541 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 6601 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 6661 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 6721 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 6781 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc 6841 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg 6901 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt 6961 gaccgacaat tgcatgaaga atctgcttag g //