LOCUS V023786 6509 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023786 VERSION V023786 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..1999 /label="PNMA1(NM_006029)" /note="PNMA1(NM_006029)" /gene="PNMA1" CDS 2006..2071 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2109..2333 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2379..2807 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2821..3150 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3217..4011 /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 4185..4306 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4355..4371) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4379..4395 /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(4403..4433) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4448..4469 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4757..5342) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5513..6373) /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(6374..6478) /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 atggcgatga cactgttgga 961 agactggtgc cgggggatgg atgtgaactc ccagagagct ctgttagtct ggggcatccc 1021 agtgaactgt gatgaggctg aaatcgaaga gaccctccag gctgcgatgc cccaggtctc 1081 ctaccgaatg cttgggagaa tgttctggag ggaagaaaat gcgaaagcag ccttattaga 1141 gctcactggc gctgtagatt acgccgcgat ccccagggag atgccgggca aaggaggggt 1201 ctggaaagtg ttatttaagc ccccaacttc tgatgctgaa tttttagaaa gattgcacct 1261 cttcctagct agagaggggt ggaccgtgca agatgttgcc cgtgtccttg ggtttcagaa 1321 ccctactccg accccgggcc cagagatgcc agcagagatg ctaaactata ttttggataa 1381 tgttattcag cctcttgttg agtccatatg gtacaagagg ctgacacttt tctcggggag 1441 ggacatccca gggcctggag aggaaacctt tgatccctgg ctggagcaca ctaatgaggt 1501 cctagaggag tggcaggtgt ccgatgtaga aaagaggcgg cggttgatgg agagtcttag 1561 aggccccgcc gctgatgtta ttcgcatcct taagtccaac aaccccgcga taaccactgc 1621 cgaatgcctg aaggcgcttg agcaggtgtt tgggagcgtt gagagctcta gggatgccca 1681 gatcaaattt ctgaacactt atcagaaccc gggagaaaaa ttgtctgctt atgtcattcg 1741 tctggagcct ctgctacaga aggtggtaga gaagggggcc attgataaag ataatgtgaa 1801 ccaggcccgc ctagagcagg tcattgccgg ggccaaccac agcggggcca tccgaaggca 1861 gctgtggctt accggggctg gggaagggcc agccccaaac ctctttcagt tgctggtgca 1921 gatccgtgag gaggaagcca aggaggagga ggaggaggct gaggccaccc ttctgcagtt 1981 aggcctggaa gggcacttcc tcgaggacta caaagaccat gacggtgatt ataaagatca 2041 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc 2101 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc 2161 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc 2221 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg 2281 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga 2341 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 2401 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 2461 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 2521 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 2581 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 2641 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 2701 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 2761 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 2821 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 2881 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 2941 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 3001 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 3061 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 3121 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc 3181 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 3241 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 3301 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 3361 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 3421 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 3481 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 3541 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 3601 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 3661 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 3721 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 3781 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 3841 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 3901 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 3961 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 4021 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 4081 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 4141 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag 4201 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 4261 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac 4321 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 4381 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 4441 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 4501 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 4561 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 4621 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 4681 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 4741 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 4801 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 4861 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 4921 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 4981 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 5041 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 5101 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 5161 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 5221 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 5281 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5341 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 5401 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 5461 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 5521 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 5581 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 5641 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 5701 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 5761 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 5821 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 5881 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 5941 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 6001 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 6061 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 6121 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 6181 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 6241 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 6301 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 6361 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 6421 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 6481 catttccccg aaaagtgcca cctgacgtc //