LOCUS V032748 7337 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032748 VERSION V032748 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..2869 /label="HSPA6(NM_002155)" /note="HSPA6(NM_002155)" /gene="HSPA6" CDS 2876..2899 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" polyA_signal 2937..3161 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3207..3635 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3649..3978 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4045..4839 /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 5013..5134 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5183..5199) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5207..5223 /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(5231..5261) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5276..5297 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5585..6170) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6341..7201) /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(7202..7306) /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 atgcaggccc cacgggagct 961 cgcggtgggc atcgacctgg gcaccaccta ctcgtgcgtg ggcgtgtttc agcagggccg 1021 cgtggagatc ctggccaacg accagggcaa ccgcaccacg cccagctacg tggccttcac 1081 cgacaccgag cggctggtcg gggacgcggc caagagccag gcggccctga acccccacaa 1141 caccgtgttc gatgccaagc ggctgatcgg gcgcaagttc gcggacacca cggtgcagtc 1201 ggacatgaag cactggccct tccgggtggt gagcgagggc ggcaagccca aggtgcgcgt 1261 atgctaccgc ggggaggaca agacgttcta ccccgaggag atctcgtcca tggtgctgag 1321 caagatgaag gagacggccg aggcgtacct gggccagccc gtgaagcacg cagtgatcac 1381 cgtgcccgcc tatttcaatg actcgcagcg ccaggccacc aaggacgcgg gggccatcgc 1441 ggggctcaac gtgttgcgga tcatcaatga gcccacggca gctgccatcg cctatgggct 1501 ggaccggcgg ggcgcgggag agcgcaacgt gctcattttt gacctgggtg ggggcacctt 1561 cgatgtgtcg gttctctcca ttgacgctgg tgtctttgag gtgaaagcca ctgctggaga 1621 tacccacctg ggaggagagg acttcgacaa ccggctcgtg aaccacttca tggaagaatt 1681 ccggcggaag catgggaagg acctgagcgg gaacaagcgt gccctgcgca ggctgcgcac 1741 agcctgtgag cgcgccaagc gcaccctgtc ctccagcacc caggccaccc tggagataga 1801 ctccctgttc gagggcgtgg acttctacac gtccatcact cgtgcccgct ttgaggaact 1861 gtgctcagac ctcttccgca gcaccctgga gccggtggag aaggccctgc gggatgccaa 1921 gctggacaag gcccagattc atgacgtcgt cctggtgggg ggctccacac gcatccccaa 1981 ggtgcagaag ttgctgcagg acttcttcaa cggcaaggag ctgaacaaga gcatcaaccc 2041 tgatgaggct gtggcctatg gggctgctgt gcaggcggcc gtgttgatgg gggacaaatg 2101 tgagaaagtg caggatctcc tgctgctgga tgtggctccc ctgtctctgg ggctggagac 2161 agcaggtggg gtgatgacca cgctgatcca gaggaacgcc actatcccca ccaagcagac 2221 ccagactttc accacctact cggacaacca gcctggggtc ttcatccagg tgtatgaggg 2281 tgagagggcc atgaccaagg acaacaacct gctggggcgt tttgaactca gtggcatccc 2341 tcctgcccca cgtggagtcc cccagataga ggtgactttt gacattgatg ctaatggcat 2401 cctgagcgtg acagccactg acaggagcac aggtaaggct aacaagatca ccatcaccaa 2461 tgacaagggc cggctgagca aggaggaggt ggagaggatg gttcatgaag ccgagcagta 2521 caaggctgag gatgaggccc agagggacag agtggctgcc aaaaactcgc tggaggccca 2581 tgtcttccat gtgaaaggtt ctttgcaaga ggaaagcctt agggacaaga ttcccgaaga 2641 ggacaggcgc aaaatgcaag acaagtgtcg ggaagtcctt gcctggctgg agcacaacca 2701 gctggcagag aaggaggagt atgagcatca gaagagggag ctggagcaaa tctgtcgccc 2761 catcttctcc aggctctatg gggggcctgg tgtccctggg ggcagcagtt gtggcactca 2821 agcccgccag ggggacccca gcaccggccc catcattgag gaggttgatc tcgaggatta 2881 caaggatgac gacgataagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt 2941 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 3001 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 3061 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 3121 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 3181 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 3241 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 3301 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 3361 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 3421 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 3481 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 3541 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 3601 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 3661 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3721 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3781 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 3841 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 3901 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 3961 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 4021 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 4081 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 4141 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 4201 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 4261 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 4321 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 4381 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 4441 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 4501 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 4561 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 4621 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 4681 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 4741 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 4801 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 4861 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 4921 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 4981 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 5041 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 5101 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 5161 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 5221 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 5281 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 5341 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5401 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 5461 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 5521 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 5581 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 5641 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 5701 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 5761 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 5821 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 5881 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 5941 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 6001 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 6061 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 6121 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 6181 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 6241 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 6301 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 6361 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 6421 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 6481 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 6541 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 6601 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 6661 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 6721 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 6781 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 6841 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 6901 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 6961 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 7021 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 7081 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 7141 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 7201 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 7261 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 7321 aagtgccacc tgacgtc //