LOCUS V025677 7488 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025677 VERSION V025677 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..2193 /label="OTUD3(NM_015207)" /note="OTUD3(NM_015207)" /gene="OTUD3" polyA_signal 2247..2869 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" promoter 2911..3208 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3262..4056 /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 4708..4842 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" promoter complement(4848..4866) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(4880..4896) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4904..4920 /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(4928..4958) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4973..4994 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5282..5870) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6041..6901) /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(6902..7006) /gene="bla" /label="AmpR promoter" rep_origin 7033..7461 /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 tgtcccgaaa gcaggcggcg 1021 aagagccggc cgggcagcgg cagccggaaa gccgaggccg agcgcaagcg ggacgagcgg 1081 gcggcgcgcc gggccctggc caaggagcgg cggaatcggc cggagtctgg cggcggcggc 1141 ggctgcgagg aggagttcgt cagcttcgcc aaccagctgc aggccctggg gctgaagctg 1201 cgggaggtgc cgggggacgg caattgcttg ttcagagctc ttggtgatca attggaggga 1261 cactcacgaa atcatctcaa gcacagacag gagacagtgg actacatgat aaagcagcgg 1321 gaagattttg aaccctttgt agaagatgac attccttttg agaagcatgt ggccagtttg 1381 gcaaagcctg gtacttttgc tggcaatgat gcaattgtag cctttgcaag aaatcatcag 1441 ttgaatgtag tgattcatca acttaatgcc cctttgtggc agattcgtgg tacagagaaa 1501 agcagcgtga gggagttaca catcgcatat cggtatggag agcactacga cagtgttcgg 1561 aggatcaatg acaactcaga ggcacctgca catctccaga cggattttca gatgcttcat 1621 caagatgaat caaataaaag agaaaagatc aagacaaagg gaatggactc tgaagacgac 1681 ctgagagatg aagtagagga tgctgtccag aaagtttgta atgcaactgg atgttcagat 1741 tttaatttaa tagtccagaa cctggaagct gaaaattata atattgaatc tgcaataatt 1801 gccgtgcttc ggatgaacca agggaagaga aataatgcag aagagaatct tgagcccagt 1861 ggtcgagtgc tgaagcagtg tggccctttg tgggaggagg gtggcagtgg tgccagaatc 1921 tttggaaatc agggcttaaa tgaaggcagg accgaaaaca ataaggcaca ggccagccct 1981 agtgaagaaa acaaagcaaa taaaaaccag ctcgcaaagg tcacaaacaa acagaggcga 2041 gaacagcagt ggatggagaa gaagaagcgg caggaggaga ggcaccgcca caaagccctg 2101 gagagcagag gtagccacag ggacaataac agaagcgaag cagaggcgaa cacgcaggtc 2161 accttggtga agaccttcgc cgctctcaac atctgagaat tcatcgatag atctgatatc 2221 ggtaccagtc gactctagag gatcccgggt ggcatccctg tgacccctcc ccagtgcctc 2281 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt 2341 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat 2401 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa 2461 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga 2521 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct 2581 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact 2641 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa 2701 ccactgctcc cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc 2761 agacacagca taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg 2821 gcactgtcct ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc 2881 cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 2941 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 3001 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 3061 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3121 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat 3181 tccagaagta gtgaggaggc ttttttggag gaattgatca gcttgggatc tgatcaagag 3241 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 3301 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 3361 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 3421 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 3481 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 3541 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 3601 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 3661 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 3721 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 3781 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 3841 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 3901 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 3961 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 4021 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 4081 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 4141 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 4201 atctcatgct ggagttcttc gcccaccccg ggctcgatcc cctcgcgagt tggttcagct 4261 gctgcctgag gctggacgac ctcgcggagt tctaccggca gtgcaaatcc gtcggcatcc 4321 aggaaaccag cagcggctat ccgcgcatcc atgcccccga actgcaggag tggggaggca 4381 cgatggccgc tttggtcgac ccggacggga cgctcctgcg cctgatacag aacgaattgc 4441 ttgcaggcat ctcatgagtg tgtcttcccg ttttccgcct gaggtcactg cgtggatgga 4501 gcgctggcgc ctgctgcgcg acggcgagct gctcaccacc cactcgccaa gctggaaccg 4561 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc cgatcataat cagccatacc 4621 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa 4681 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa 4741 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 4801 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaattccct atagtgagtc 4861 gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 4921 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 4981 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 5041 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 5101 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 5161 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 5221 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 5281 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 5341 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 5401 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 5461 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 5521 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 5581 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 5641 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 5701 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 5761 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 5821 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 5881 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 5941 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 6001 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 6061 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 6121 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 6181 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 6241 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 6301 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 6361 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 6421 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 6481 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 6541 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 6601 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 6661 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 6721 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 6781 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 6841 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 6901 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 6961 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 7021 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 7081 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt 7141 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag 7201 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt 7261 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 7321 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt 7381 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt 7441 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaattt //