LOCUS V029953 6171 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029953 VERSION V029953 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 166..182 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 343..722 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 723..926 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 952..970 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 1029..2318 /label="Smad9(NM_019483)" /note="Smad9(NM_019483)" /gene="Smad9" CDS 2337..2366 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 2385..2408 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" primer_bind complement(2428..2444) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 2455..3077 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(3226..3814) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 4143..4190 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(4422..5216) /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" promoter complement(5251..5608) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(5610..5714) /gene="bla" /label="AmpR promoter" rep_origin 5741..6169 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag 61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt 241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata 301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta 361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat 781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac 901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac 961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg 1021 cgatcgccat gcaccccagc acccccatca gctccctctt ctccttcacc agccccgcag 1081 tgaagcggct gctgggctgg aagcagggag atgaagagga gaagtgggca gagaaggcgg 1141 tggactcttt ggtgaagaag ttaaagaaga agaaaggcgc catggatgaa ctggagaggg 1201 cgctgagctg cccgggtcag cctagcaagt gtgtcaccat cccacggtcc ctcgatggac 1261 gcctccaggt gtcccaccga aaggggctgc cccacgtcat ctactgccgc gtgtggcgct 1321 ggccagacct gcagtcccat catgagctga agcccttgga gtgctgtgag ttcccgttcg 1381 gctccaagca gaaggaggtc tgcatcaacc cataccatta ccgcagagtg gagaccccag 1441 ttctgcctcc agtgctggta ccaagacaca gcgagtacaa ccctcagctc agcctcctgg 1501 ccaagttccg aagtgcctcg ctgcacagcg aacccctcat gccgcacaac gccacctacc 1561 ctgactcttt ccagcagtct ctctgtccgg caccgccctc ctcgccaggc catgtgtttc 1621 cgcagtctcc atgccccacc agctacccgc actcccccgg aagtccttcc gagtcagaca 1681 gtccctatca acactcagac ttccggccag tttgctacga ggaaccccag cactggtgtt 1741 ctgttgccta ctacgaacta aacaaccggg tcggagagac tttccaggcg tcctcgcgga 1801 gcgtgctcat agacggcttc accgaccctt ccaataacag gaataggttc tgccttgggc 1861 ttctctcaaa tgtaaacaga aactcgacca tagaaaacac caggaggcac attggaaagg 1921 gtgtgcattt gtactacgtt gggggcgagg tgtatgcgga gtgcgtgagc gacagcagca 1981 tctttgtcca gagccggaac tgcaactacc agcacggctt ccacccggcc accgtctgca 2041 agatccccag cggctgcagc ctcaaggtct tcaacaacca gctcttcgcc cagctgctcg 2101 cccagtccgt gcaccacggc tttgaagtgg tgtatgagct gacgaagatg tgcacgattc 2161 ggatgagctt tgtgaagggc tggggagcag agtatcatcg ccaggatgtc acgagcaccc 2221 cctgctggat cgagatccat cttcatggac cgctgcagtg gttggataag gtgctcactc 2281 agatgggctc cccacacaac cctatctctt cagtgtctac gcgtacgcgg ccgctcgagc 2341 agaaactcat ctcagaagag gatctggcag caaatgatat cctggattac aaggatgacg 2401 acgataaggt ttaaacggcc ggccgcggtc atagctgttt cctgaacaga tcccgggtgg 2461 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc 2521 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat 2581 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt 2641 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact 2701 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga 2761 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc 2821 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct 2881 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta 2941 aaataactat accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa 3001 ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag 3061 tagatgcctg ttgaattggg tacgcggcca gcggcgagcg gtatcagctc actcaaaggc 3121 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 3181 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 3241 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 3301 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 3361 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 3421 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 3481 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 3541 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 3601 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 3661 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 3721 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 3781 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 3841 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 3901 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 3961 atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg 4021 gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc 4081 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt 4141 atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg 4201 ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa 4261 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat 4321 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt 4381 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga 4441 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag 4501 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc 4561 tgatagcgat ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt 4621 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg 4681 ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg 4741 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga 4801 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt 4861 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc 4921 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca 4981 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt 5041 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac 5101 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat 5161 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga 5221 aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc 5281 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa 5341 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat 5401 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact 5461 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat 5521 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac 5581 attccacagc tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt 5641 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 5701 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca 5761 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 5821 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 5881 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 5941 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 6001 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 6061 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 6121 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt t //