LOCUS V047187 10106 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047187 VERSION V047187 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 951..4913 /label="KDM3A(NM_018433)" /note="KDM3A(NM_018433)" /gene="KDM3A" CDS 4944..5639 /codon_start=1 /note="" /product="promiscuous R40G mutant of a biotin protein ligase from Aquifex aeolicus (Kim et al., 2016)" /transl_table=1 /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI LSGEFSLRRS" /label="BioID2" CDS 5640..5666 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" polyA_signal 5702..5926 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 5972..6400 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 6414..6744 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 6811..7605 /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 7779..7900 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(7949..7965) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7973..7989 /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(7997..8027) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8042..8063 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8351..8939) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9110..9970) /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(9971..10075) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttaaggcc tgttaaccgg tcgtacgtct ccggattcga attcgccacc atggtgctca 961 cgctcggaga aagttggccg gtattggtgg ggaggaggtt tctcagtctg tccgcagccg 1021 acggcagcga tggcagccac gacagctggg acgtggagcg cgtcgccgag tggccctggc 1081 tctccgggac cattcgagct gtttcccaca ccgacgttac caagaaggat ctgaaggtgt 1141 gtgtggaatt tgatggggaa tcttggagga aaagaagatg gatagaagtc tacagccttc 1201 taaggagagc atttttagta gaacataatt tggttttagc tgaacgaaag tcacctgaaa 1261 tttctgaacg aattgtacag tggcctgcaa taacgtacaa acctctgttg gacaaagctg 1321 gtttgggatc cataacttct gttcgctttc tgggagatca acaaagagta tttctttcta 1381 aagacctttt gaagcctata caggatgtaa acagtcttcg actttctctt acggataatc 1441 agattgtcag taaagaattt caagctttga ttgtgaagca tttagatgaa agccatcttt 1501 taaaaggtga caaaaactta gttggttcag aagtaaaaat ttatagcttg gacccatcta 1561 ctcagtggtt ttcagcaacc gttataaatg gaaacccagc atcaaaaact cttcaagtca 1621 actgtgagga gattccagca ctgaaaattg ttgatccgtc actgattcat gttgaagttg 1681 tacacgataa ccttgtgaca tgtggtaatt ctgcaagaat tggagctgta aaacgcaagt 1741 cttctgagaa taatggaacc ctggtttcca aacaagcaaa atcttgctct gaggcctctc 1801 ccagtatgtg tcctgtgcag tctgtaccta caacagtttt taaggagata ctgcttggct 1861 gtactgcggc aactccacct agtaaggacc caagacagca aagtactccc caggctgcca 1921 actctccacc taaccttgga gcaaaaattc ctcaaggatg tcataaacaa agtttaccag 1981 aggaaatttc ttcctgtcta aatacaaagt ctgaagctct gagaacaaaa ccagatgtct 2041 gcaaagcagg gttgctctca aagtcctctc agattggaac tggagacttg aaaattctga 2101 ctgagccaaa aggcagctgt actcagccta agacaaacac tgatcaggaa aacagattgg 2161 agtctgttcc acaagcattg actggccttc ctaaggagtg cttacctaca aaggcttctt 2221 ctaaggcaga attggaaatt gccaatcctc ctgaactgca gaagcaccta gaacatgcac 2281 cttccccatc ggatgtttca aatgcaccag aagtgaaagc aggtgtcaat agtgatagcc 2341 ctaataactg ttcaggaaaa aaggtagaac cttcagcttt agcttgccga tcacagaatt 2401 taaaggaatc ttcagtaaaa gtagataatg aaagctgttg ttcaagaagc aacaataaaa 2461 tccagaatgc cccatccagg aagtcggttt tgacagaccc agctaaactc aaaaagctgc 2521 aacagagtgg cgaggccttc gtacaggatg attcttgtgt gaacatcgtg gcacagttgc 2581 ctaaatgccg agagtgtcgc ttggacagtc tccgcaagga taaggagcaa cagaaggact 2641 cacctgtgtt ttgccgcttc tttcacttca ggaggttaca attcaacaaa catggtgtgt 2701 tgcgggtaga aggcttctta acaccaaaca agtatgacaa tgaagcaatt ggcttgtggt 2761 tacctttaac caaaaacgtt gtggggattg atttggacac agcaaagtac atcttggcca 2821 acattggaga ccacttctgt caaatggtga tttctgaaaa ggaagctatg tcaactattg 2881 agccacacag acaggttgct tggaagcgag ctgtcaaagg tgttcgagaa atgtgtgatg 2941 tgtgcgacac caccatcttc aacctgcact gggtgtgtcc tcggtgtggg tttggagtat 3001 gtgtggactg ctaccggatg aagagaaaga attgccaaca gggtgctgct tacaagactt 3061 tctcttggct aaaatgtgtg aagagtcaga tacatgaacc agagaactta atgcccacac 3121 agatcattcc tggaaaagca ctctatgatg ttggagacat tgttcattct gtaagagcga 3181 aatggggaat aaaggcaaac tgcccttgtt caaacaggca attcaaactc ttttcaaagc 3241 cagcctcaaa ggaagaccta aaacagactt ctttagctgg agaaaaaccg actcttggtg 3301 cagtgctcca gcagaatccc tcagtgttgg agccagcagc tgtgggtggg gaagcagcct 3361 ccaagccagc cggcagcatg aagcctgcct gtccagccag cacatctcct ctaaactggc 3421 tggccgacct aaccagcggg aatgtcaaca aggaaaacaa ggaaaaacaa ccaacaatgc 3481 caattttaaa gaatgaaatc aaatgccttc cacccctccc acctttaagc aaatccagca 3541 cagtcctcca tacgtttaac agcacaattt tgacacccgt aagcaacaac aattctggtt 3601 tcctccggaa tctcttgaat tcttctacag gaaagacaga aaatggactc aagaatacac 3661 caaaaatcct tgatgacatc tttgcctctt tggtgcaaaa taagacgact tctgatttat 3721 ctaagaggcc tcaaggacta accatcaagc ccagcattct gggctttgac actcctcact 3781 attggctttg tgataatcgc ttgctgtgct tgcaagaccc caacaataag agcaactgga 3841 atgtgtttag ggagtgctgg aaacaagggc agccagtgat ggtgtctgga gtgcatcata 3901 aattgaactc tgaactttgg aaacctgaat ccttcaggaa agagtttggt gagcaggaag 3961 tagacctagt taattgtagg accaatgaaa tcatcacagg agccacagta ggagacttct 4021 gggatggatt tgaagatgtt ccaaatcgtt tgaaaaatga aaaagaacca atggtgttga 4081 aacttaagga ctggccacca ggagaagatt ttagagatat gatgccttcc aggtttgatg 4141 atctgatggc caacattcca ctgcccgagt acacaaggcg agatggcaaa ctgaatttgg 4201 cctctaggct gccaaactac tttgttcggc cagatctggg ccccaagatg tataatgctt 4261 atggattaat cactcctgaa gatcggaaat atggaacaac aaatcttcac ttagatgtat 4321 ctgatgcagc taatgtcatg gtctatgtgg gaattcccaa aggacagtgt gagcaagaag 4381 aagaagtcct taagaccatc caagatggag attctgacga actcacaata aagcgattta 4441 ttgaaggaaa agagaagcca ggagcactgt ggcacatata tgctgcaaag gacacggaga 4501 agataaggga atttcttaaa aaggtatcag aagagcaagg tcaagaaaac ccagcagacc 4561 acgatcctat tcatgatcaa agctggtatt tagaccgatc attaagaaaa cgtcttcatc 4621 aagagtatgg agttcaaggc tgggctattg tacagtttct tggggatgtg gtgtttatcc 4681 cggcaggagc tccacatcag gttcataact tatatagctg catcaaagtg gctgaagatt 4741 ttgtttctcc agagcatgtt aaacactgct tctggcttac tcaggaattc cgatatctgt 4801 cacagactca taccaatcac gaagataaat tacaggtgaa gaatgttatc taccatgcag 4861 tgaaagatgc agttgctatg ctgaaagcca gtgaatccag ttttggcaaa cctggtggag 4921 gcggatcagg aggcggtgga tccttcaaga acctgatctg gctgaaggag gtggacagca 4981 cccaggagag actgaaggag tggaacgtga gctacggcac cgccctggtg gccgacagac 5041 agaccaaggg cagaggcggc ctgggcagaa agtggctgag ccaggagggc ggcctgtact 5101 tcagcttcct gctgaacccc aaggagttcg agaacctgct gcagctgccc ctggtgctgg 5161 gcctgagcgt gagcgaggcc ctggaggaga tcaccgagat ccccttcagc ctgaagtggc 5221 ccaacgacgt gtacttccag gagaagaagg tgagcggcgt gctgtgcgag ctgagcaagg 5281 acaagctgat cgtgggcatc ggcatcaacg tgaaccagag agagatcccc gaggagatca 5341 aggacagagc caccaccctg tacgagatca ccggcaagga ctgggacaga aaggaggtgc 5401 tgctgaaggt gctgaagaga atcagcgaga acctgaagaa gttcaaggag aagagcttca 5461 aggagttcaa gggcaagatc gagagcaaga tgctgtacct gggcgaggag gtgaagctgc 5521 tgggcgaggg caagatcacc ggcaagctgg tgggcctgag cgagaagggc ggcgccctga 5581 tcctgaccga ggagggcatc aaggagatcc tgagcggcga gttcagcctg agaagaagct 5641 acccttacga tgtaccggat tacgcataga agcttaagtt taaaccgctg atcagcctcg 5701 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 5761 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 5821 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 5881 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 5941 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 6001 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 6061 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 6121 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 6181 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 6241 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 6301 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg 6361 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 6421 agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat 6481 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg 6541 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg 6601 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 6661 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt 6721 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct 6781 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt 6841 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc 6901 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag 6961 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg 7021 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac 7081 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc 7141 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc 7201 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc 7261 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg 7321 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat 7381 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc 7441 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa 7501 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat 7561 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt 7621 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg 7681 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc 7741 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata 7801 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 7861 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga 7921 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 7981 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 8041 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 8101 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 8161 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 8221 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 8281 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 8341 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 8401 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 8461 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 8521 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg 8581 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 8641 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 8701 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 8761 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 8821 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 8881 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 8941 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 9001 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 9061 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 9121 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 9181 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 9241 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 9301 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 9361 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 9421 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 9481 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 9541 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 9601 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 9661 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 9721 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 9781 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 9841 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 9901 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 9961 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 10021 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 10081 ttccccgaaa agtgccacct gacgtc //