LOCUS V036783 7388 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036783 VERSION V036783 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 941..2878 /label="HSPA8(NM_006597)" /note="HSPA8(NM_006597)" /gene="HSPA8" CDS 2885..2950 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2988..3212 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3258..3686 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3700..4029 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4096..4890 /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 5064..5185 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5234..5250) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5258..5274 /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(5282..5312) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5327..5348 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5636..6221) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6392..7252) /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(7253..7357) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgtccaagg gacctgcagt 961 tggtattgat cttggcacca cctactcttg tgtgggtgtt ttccagcacg gaaaagtcga 1021 gataattgcc aatgatcagg gaaaccgaac cactccaagc tatgtcgcct ttacggacac 1081 tgaacggttg atcggtgatg ccgcaaagaa tcaagttgca atgaacccca ccaacacagt 1141 ttttgatgcc aaacgtctga ttggacgcag atttgatgat gctgttgtcc agtctgatat 1201 gaaacattgg ccctttatgg tggtgaatga tgctggcagg cccaaggtcc aagtagaata 1261 caagggagag accaaaagct tctatccaga ggaggtgtct tctatggttc tgacaaagat 1321 gaaggaaatt gcagaagcct accttgggaa gactgttacc aatgctgtgg tcacagtgcc 1381 agcttacttt aatgactctc agcgtcaggc taccaaagat gctggaacta ttgctggtct 1441 caatgtactt agaattatta atgagccaac tgctgctgct attgcttacg gcttagacaa 1501 aaaggttgga gcagaaagaa acgtgctcat ctttgacctg ggaggtggca cttttgatgt 1561 gtcaatcctc actattgagg atggaatctt tgaggtcaag tctacagctg gagacaccca 1621 cttgggtgga gaagattttg acaaccgaat ggtcaaccat tttattgctg agtttaagcg 1681 caagcataag aaggacatca gtgagaacaa gagagctgta agacgcctcc gtactgcttg 1741 tgaacgtgct aagcgtaccc tctcttccag cacccaggcc agtattgaga tcgattctct 1801 ctatgaagga atcgacttct atacctccat tacccgtgcc cgatttgaag aactgaatgc 1861 tgacctgttc cgtggcaccc tggacccagt agagaaagcc cttcgagatg ccaaactaga 1921 caagtcacag attcatgata ttgtcctggt tggtggttct actcgtatcc ccaagattca 1981 gaagcttctc caagacttct tcaatggaaa agaactgaat aagagcatca accctgatga 2041 agctgttgct tatggtgcag ctgtccaggc agccatcttg tctggagaca agtctgagaa 2101 tgttcaagat ttgctgctct tggatgtcac tcctctttcc cttggtattg aaactgctgg 2161 tggagtcatg actgtcctca tcaagcgtaa taccaccatt cctaccaagc agacacagac 2221 cttcactacc tattctgaca accagcctgg tgtgcttatt caggtttatg aaggcgagcg 2281 tgccatgaca aaggataaca acctgcttgg caagtttgaa ctcacaggca tacctcctgc 2341 accccgaggt gttcctcaga ttgaagtcac ttttgacatt gatgccaatg gtatactcaa 2401 tgtctctgct gtggacaaga gtacgggaaa agagaacaag attactatca ctaatgacaa 2461 gggccgtttg agcaaggaag acattgaacg tatggtccag gaagctgaga agtacaaagc 2521 tgaagatgag aagcagaggg acaaggtgtc atccaagaat tcacttgagt cctatgcctt 2581 caacatgaaa gcaactgttg aagatgagaa acttcaaggc aagattaacg atgaggacaa 2641 acagaagatt ctggacaagt gtaatgaaat tatcaactgg cttgataaga atcagactgc 2701 tgagaaggaa gaatttgaac atcaacagaa agagctggag aaagtttgca accccatcat 2761 caccaagctg taccagagtg caggaggcat gccaggagga atgcctgggg gatttcctgg 2821 tggtggagct cctccctctg gtggtgcttc ctcagggccc accattgaag aggttgatct 2881 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga 2941 cgatgacaag tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag 3001 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 3061 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 3121 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 3181 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg 3241 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 3301 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 3361 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 3421 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 3481 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 3541 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 3601 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 3661 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga 3721 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3781 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 3841 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 3901 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 3961 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 4021 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga 4081 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 4141 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 4201 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 4261 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 4321 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 4381 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 4441 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 4501 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 4561 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 4621 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 4681 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 4741 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 4801 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 4861 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 4921 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 4981 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 5041 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa 5101 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 5161 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 5221 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 5281 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 5341 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 5401 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 5461 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 5521 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 5581 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 5641 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 5701 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 5761 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 5821 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 5881 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 5941 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 6001 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 6061 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 6121 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg 6181 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 6241 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 6301 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 6361 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 6421 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 6481 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 6541 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 6601 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 6661 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 6721 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 6781 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 6841 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 6901 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 6961 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 7021 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 7081 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 7141 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 7201 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 7261 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 7321 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 7381 ctgacgtc //