LOCUS V022435 7925 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022435 VERSION V022435 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..2605 /label="HSPA1A(NM_005345)" /note="HSPA1A(NM_005345)" /gene="HSPA1A" CDS 2612..2677 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2701..3289 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3355..3403 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3605..4033 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4047..4376 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4443..5237 /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 5413..5534 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5583..5599) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5607..5623 /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(5631..5661) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5676..5697 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5985..6573) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6744..7604) /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(7605..7709) /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 ccatggccaa agccgcggcg atcggcatcg acctgggcac 721 cacctactcc tgcgtggggg tgttccaaca cggcaaggtg gagatcatcg ccaacgacca 781 gggcaaccgc accaccccca gctacgtggc cttcacggac accgagcggc tcatcgggga 841 tgcggccaag aaccaggtgg cgctgaaccc gcagaacacc gtgtttgacg cgaagcggct 901 gattggccgc aagttcggcg acccggtggt gcagtcggac atgaagcact ggcctttcca 961 ggtgatcaac gacggagaca agcccaaggt gcaggtgagc tacaaggggg agaccaaggc 1021 attctacccc gaggagatct cgtccatggt gctgaccaag atgaaggaga tcgccgaggc 1081 gtacctgggc tacccggtga ccaacgcggt gatcaccgtg ccggcctact tcaacgactc 1141 gcagcgccag gccaccaagg atgcgggtgt gatcgcgggg ctcaacgtgc tgcggatcat 1201 caacgagccc acggccgccg ccatcgccta cggcctggac agaacgggca agggggagcg 1261 caacgtgctc atctttgacc tgggcggggg caccttcgac gtgtccatcc tgacgatcga 1321 cgacggcatc ttcgaggtga aggccacggc cggggacacc cacctgggtg gggaggactt 1381 tgacaacagg ctggtgaacc acttcgtgga ggagttcaag agaaaacaca agaaggacat 1441 cagccagaac aagcgagccg tgaggcggct gcgcaccgcc tgcgagaggg ccaagaggac 1501 cctgtcgtcc agcacccagg ccagcctgga gatcgactcc ctgtttgagg gcatcgactt 1561 ctacacgtcc atcaccaggg cgaggttcga ggagctgtgc tccgacctgt tccgaagcac 1621 cctggagccc gtggagaagg ctctgcgcga cgccaagctg gacaaggccc agattcacga 1681 cctggtcctg gtcgggggct ccacccgcat ccccaaggtg cagaagctgc tgcaggactt 1741 cttcaacggg cgcgacctga acaagagcat caaccccgac gaggctgtgg cctacggggc 1801 ggcggtgcag gcggccatcc tgatggggga caagtccgag aacgtgcagg acctgctgct 1861 gctggacgtg gctcccctgt cgctggggct ggagacggcc ggaggcgtga tgactgccct 1921 gatcaagcgc aactccacca tccccaccaa gcagacgcag atcttcacca cctactccga 1981 caaccaaccc ggggtgctga tccaggtgta cgagggcgag agggccatga cgaaagacaa 2041 caatctgttg gggcgcttcg agctgagcgg catccctccg gcccccaggg gcgtgcccca 2101 gatcgaggtg accttcgaca tcgatgccaa cggcatcctg aacgtcacgg ccacggacaa 2161 gagcaccggc aaggccaaca agatcaccat caccaacgac aagggccgcc tgagcaagga 2221 ggagatcgag cgcatggtgc aggaggcgga gaagtacaaa gcggaggacg aggtgcagcg 2281 cgagagggtg tcagccaaga acgccctgga gtcctacgcc ttcaacatga agagcgccgt 2341 ggaggatgag gggctcaagg gcaagatcag cgaggcggac aagaagaagg tgctggacaa 2401 gtgtcaagag gtcatctcgt ggctggacgc caacaccttg gccgagaagg acgagtttga 2461 gcacaagagg aaggagctgg agcaggtgtg taaccccatc atcagcggac tgtaccaggg 2521 tgccggtggt cccgggcctg ggggcttcgg ggctcagggt cccaagggag ggtctgggtc 2581 aggccccacc attgaggagg tagatctcga ggactacaaa gaccatgacg gtgattataa 2641 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac 2701 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 2761 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 2821 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 2881 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 2941 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 3001 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 3061 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 3121 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 3181 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 3241 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg 3301 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa 3361 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg 3421 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 3481 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 3541 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat 3601 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 3661 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 3721 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga 3781 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 3841 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 3901 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 3961 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 4021 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 4081 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 4141 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 4201 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 4261 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 4321 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 4381 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 4441 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 4501 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 4561 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 4621 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 4681 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 4741 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 4801 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 4861 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 4921 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 4981 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 5041 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 5101 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 5161 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 5221 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc 5281 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 5341 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 5401 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 5461 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 5521 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 5581 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 5641 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 5701 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 5761 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 5821 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 5881 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 5941 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 6001 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 6061 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 6121 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa 6181 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 6241 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 6301 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 6361 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 6421 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 6481 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 6541 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 6601 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 6661 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 6721 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 6781 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 6841 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 6901 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 6961 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 7021 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 7081 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 7141 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 7201 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 7261 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 7321 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 7381 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 7441 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 7501 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 7561 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 7621 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 7681 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 7741 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 7801 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 7861 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 7921 ttagg //