LOCUS V025703 10350 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025703 VERSION V025703 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 141..157 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 318..697 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 698..901 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 931..996 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 1000..5055 /label="USP31(NM_020718)" /note="USP31(NM_020718)" /gene="USP31" polyA_signal 5109..5731 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" promoter 5773..6070 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 6124..6918 /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 7570..7704 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" promoter complement(7710..7728) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(7742..7758) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7766..7782 /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(7790..7820) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7835..7856 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8144..8732) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8903..9763) /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(9764..9868) /gene="bla" /label="AmpR promoter" rep_origin 9895..10323 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa 181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta 241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg 361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa 961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tgtccaaggt aacggcgcct 1021 gggtccgggc cgccggcggc ggcgagcggg aaggagaagc gctccttcag caagcggctg 1081 tttcggagcg gccgcgctgg cggcggcggc gcggggggcc ccggggcgtc cgggccggcc 1141 gcgccttcct cgccctcctc gccctcctct gcacgctcgg tgggcagctt catgagccgc 1201 gttctcaaga cgctgtccac gctctcgcac cttagctctg agggcgccgc cccagaccgc 1261 ggcggcctcc gcagctgctt cccgcccggg ccggccgccg cgcccacgcc gccgccgtgc 1321 ccgccgccgc ccgcctctcc cgcgccgccc gcttgcgccg ccgagccggt gcccggcgtg 1381 gcggggctcc gcaaccacgg caacacgtgc ttcatgaacg ccacgctgca gtgcctcagc 1441 aacaccgagc tcttcgccga gtacctggcg ctgggccagt accgggcggg gcggcccgag 1501 ccctcgcctg acccggagca gcctgcgggc cgcggcgcgc agggccaggg cgaggtcact 1561 gagcagctgg cgcacctggt gcgggccctc tggaccctgg agtacacccc gcagcacagc 1621 cgcgacttca agactattgt gtcaaagaat gcactgcagt accggggaaa ttcccaacat 1681 gatgcccagg agtttctgct gtggcttttg gaccgagttc atgaagacct caaccattca 1741 gtgaagcaga gtggccagcc tcctctgaag ccaccatcag agactgatat gatgcctgag 1801 ggaccatctt tccctgtctg tagcactttt gtacaagaac tctttcaagc gcaatacaga 1861 tcttctttga cgtgtcctca ttgtcagaaa cagagcaaca cttttgatcc tttcctttgc 1921 atttctttgc caattcctct gccccacaca aggcctctct atgtcactgt agtgtatcaa 1981 ggcaaatgtt ctcactgcat gaggattggt gtggccgtac ctctgtctgg gactgtcgcc 2041 agacttcggg aagcagtgtc tatggaaaca aagatcccca ctgatcagat tgtgttaaca 2101 gaaatgtact atgatgggtt ccatcgttcc ttttgtgata cagacgacct ggaaacagtc 2161 catgaaagcg actgcatttt tgcctttgag actcccgaaa tatttaggcc tgaaggaatt 2221 ctcagtcaaa gaggaattca tttaaacaac aacctaaacc acttgaaatt tggcttggat 2281 tatcatagac tgtcttctcc tacacaaaca gcagcaaagc aggggaaaat ggattctccc 2341 acatcaagag caggcagcga caagattgtc ctgttggtgt gtaaccgagc ctgcactggg 2401 caacaaggga aaagatttgg actgcctttt gtgctgcact tagagaagac aatagcttgg 2461 gaccttctgc agaaggaaat cttggagaag atgaagtatt tcttgaggcc cacggtttgc 2521 attcaggtgt gtccattcag cttgcgtgtg gtcagtgttg ttggaataac atatttgctg 2581 ccccaggagg agcagccctt gtgccaccca atagtagaaa gggcattaaa atcttgtgga 2641 ccaggtggca ctgctcatgt gaaattagta gtcgagtggg acaaggagac aagagatttc 2701 ttatttgtaa atactgagga tgagtatatt cctgatgcag aaagtgttcg tctgcaaagg 2761 gagcgtcatc atcagcctca aacctgcact ttatcccagt gtttccaact gtacaccaaa 2821 gaggagcggc ttgcccccga tgatgcctgg cgttgcccac actgtaagca gctgcagcag 2881 ggaagcatta cgttaagcct ctggactctg cctgatgtgc ttattataca tctaaagaga 2941 tttcggcagg aaggagacag gcgcatgaaa cttcagaaca tggtcaaatt ccccttgact 3001 ggcctggaca tgacacctca cgtggttaag aggagccaga gcagctggag tttgccatcg 3061 cattggtccc cgtggagacg gccctatgga ctcgggaggg accctgagga ctacatctat 3121 gacctgtatg ctgtgtgcaa tcaccatggc accatgcaag gggggcacta cacagcgtac 3181 tgtaagaact ctgtggacgg cctctggtac tgcttcgatg acagcgatgt gcagcagctg 3241 tcagaagatg aggtctgcac gcagacagca tacatcctct tctaccagag gcggacagcc 3301 atcccgtcat ggtcagccaa cagctcggtg gcaggctcca caagttcttc cctgtgtgaa 3361 cactgggtga gccggctccc gggcagcaag ccagccagcg tgacctctgc agcttcctcc 3421 agacgcacct ccctggcgtc gctctctgag tccgtggaga tgactggaga aaggagtgaa 3481 gatgatggag gcttttcaac tcgaccattt gtgagaagtg tccagcgtca gagtttgtca 3541 tccagatctt ctgtcaccag ccccttggcc gtcaatgaaa attgcatgag accttcatgg 3601 tccctgtctg ctaagctgca gatgcgctcc aattctccat cccgattttc aggggattcg 3661 ccaattcaca gctctgcttc caccttggag aagattgggg aggcagcaga tgacaaggtc 3721 tccatctctt gctttggtag cttgcggaac ctttctagca gttaccagga accaagcgac 3781 agtcatagtc gccgtgagca caaggctgtg ggccgggccc ctctggctgt catggaaggc 3841 gtgttcaaag acgaatcgga cacccgcaga ttgaactcca gtgtcgtaga tacacagagc 3901 aaacattcag cacaagggga ccgcctgccc ccgctctctg gtccatttga taacaataat 3961 cagatcgctt atgtggatca gagcgactcc gtagacagct ctccagtcaa agaggtgaaa 4021 gcccccagcc acccaggctc actcgcaaag aaaccagaga gcacaactaa gagatccccc 4081 agttccaaag gcacttctga gccagagaaa agcttgcgga aggggagacc agccttggca 4141 agccaggagt catccctttc aagtacatcc ccttcttctc ctcttcctgt aaaagtctct 4201 ctaaagccct cccgctcccg cagcaaagca gattcttctt ccaggggcag tggacggcat 4261 tcatcccctg cccctgccca acccaaaaag gagtcatccc cgaaatctca ggactccgtg 4321 tcatctcctt cgccacagaa gcagaagtca gcctcggccc tcacctacac tgcttcctcc 4381 acatctgcca aaaaggcctc gggccctgcc acaaggagcc ctttcccacc tgggaagagc 4441 aggacttcag accacagctt gagtagagag ggctccagac aaagcttggg ttctgacaga 4501 gccagcgcca cctccacctc caaacccaat tcccctcggg tgagccaggc ccgagcaggg 4561 gagggcaggg gggccgggaa gcacgtgcgg agctcctcca tggccagcct gcgctccccc 4621 agcacaagca tcaagtctgg tttgaagagg gacagcaagt ctgaggacaa ggggctgtcc 4681 ttcttcaaat cagccttgag acagaaggaa acccggcgct cgacggatct tggcaagaca 4741 gccttgctct ctaaaaaggc tggtgggagc tctgttaagt ctgtctgtaa gaacaccggg 4801 gacgacgagg cagagagagg ccaccagcct ccagcttccc agcagccaaa tgcaaataca 4861 acgggaaaag agcagcttgt caccaaggac cctgcttctg ccaaacattc cctgctgtcc 4921 gctcgcaaat ccaagtcttc ccaactagac tctggagttc cctcgtctcc gggtggcagg 4981 cagtctgcag agaaatcctc aaaaaagtta tcttctagca tgcaaacctc tgcacggcct 5041 tctcaaaaac ctcagtgaga attcatcgat agatctgata tcggtaccag tcgactctag 5101 aggatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg 5161 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact 5221 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg 5281 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac 5341 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc 5401 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt 5461 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct 5521 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg 5581 tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac 5641 ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg 5701 ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg 5761 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 5821 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 5881 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 5941 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 6001 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 6061 gcttttttgg aggaattgat cagcttggga tctgatcaag agacaggatg aggatcgttt 6121 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 6181 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 6241 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 6301 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 6361 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 6421 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 6481 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 6541 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 6601 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 6661 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 6721 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 6781 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 6841 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 6901 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 6961 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 7021 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 7081 tcgcccaccc cgggctcgat cccctcgcga gttggttcag ctgctgcctg aggctggacg 7141 acctcgcgga gttctaccgg cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct 7201 atccgcgcat ccatgccccc gaactgcagg agtggggagg cacgatggcc gctttggtcg 7261 acccggacgg gacgctcctg cgcctgatac agaacgaatt gcttgcaggc atctcatgag 7321 tgtgtcttcc cgttttccgc ctgaggtcac tgcgtggatg gagcgctggc gcctgctgcg 7381 cgacggcgag ctgctcacca cccactcgcc aagctggaac cgtaaaaagg ccgcgttgct 7441 ggcgtttttc cataggctcc gccgatcata atcagccata ccacatttgt agaggtttta 7501 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt 7561 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 7621 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 7681 aatgtatctt atcatgtctg gatcaattcc ctatagtgag tcgtattaaa ttcgtaatca 7741 tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 7801 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 7861 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 7921 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 7981 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 8041 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 8101 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 8161 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 8221 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 8281 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 8341 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 8401 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 8461 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 8521 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 8581 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 8641 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 8701 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 8761 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 8821 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 8881 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 8941 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 9001 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 9061 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 9121 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 9181 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 9241 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 9301 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 9361 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 9421 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 9481 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 9541 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 9601 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 9661 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 9721 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 9781 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 9841 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc 9901 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 9961 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 10021 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 10081 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 10141 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 10201 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 10261 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 10321 attttaacaa aatattaacg cttacaattt //