LOCUS V029151 5574 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029151 VERSION V029151 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..1721 /label="Ascl1(NM_008553)" /note="Ascl1(NM_008553)" /gene="Ascl1" CDS 1740..1769 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 1788..1811 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" primer_bind complement(1831..1847) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 1858..2480 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(2629..3217) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 3546..3593 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(3825..4619) /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(4654..5011) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(5013..5117) /gene="bla" /label="AmpR promoter" rep_origin 5144..5572 /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 ggagagctct ggcaagatgg agagtggagc cggccagcag ccgcagcccc 1081 cgcagccctt cctgcctccc gcagcctgct tctttgcgac cgcggcggcg gcggcagcgg 1141 cggcggccgc ggcagctcag agcgcgcagc agcaacagcc gcaggcgccg ccgcagcagg 1201 cgccgcagct gagcccggtg gccgacagcc agccctcagg gggcggtcac aagtcagcgg 1261 ccaagcaggt caagcgccag cgctcgtcct ctccggaact gatgcgctgc aaacgccggc 1321 tcaacttcag cggcttcggc tacagcctgc cacagcagca gccggccgcc gtggcgcgcc 1381 gcaacgagcg cgagcgcaac cgggtcaagt tggtcaacct gggttttgcc accctccggg 1441 agcatgtccc caacggcgcg gccaacaaga agatgagcaa ggtggagacg ctgcgctcgg 1501 cggtcgagta catccgcgcg ctgcagcagc tgctggacga gcacgacgcg gtgagcgctg 1561 cctttcaggc gggcgtcctg tcgcccacca tctcccccaa ctactccaac gacttgaact 1621 ctatggcggg ttctccggtc tcgtcctact cctccgacga gggatcctac gaccctctta 1681 gcccagagga acaagagctg ctggacttta ccaactggtt cacgcgtacg cggccgctcg 1741 agcagaaact catctcagaa gaggatctgg cagcaaatga tatcctggat tacaaggatg 1801 acgacgataa ggtttaaacg gccggccgcg gtcatagctg tttcctgaac agatcccggg 1861 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg 1921 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc 1981 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc 2041 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc 2101 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg 2161 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt 2221 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg 2281 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt 2341 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc 2401 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact 2461 cagtagatgc ctgttgaatt gggtacgcgg ccagcggcga gcggtatcag ctcactcaaa 2521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 2581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 2641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 2701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 2761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 2821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 2881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 2941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 3001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 3061 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 3121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 3181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 3241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 3301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 3361 tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc 3421 tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg 3481 gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg 3541 tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc 3601 gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc 3661 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac 3721 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag 3781 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca 3841 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg 3901 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg 3961 tcctgatagc gatccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca 4021 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg 4081 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct 4141 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg 4201 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc 4261 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc 4321 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc 4381 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc 4441 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct 4501 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg 4561 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg 4621 cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa 4681 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat 4741 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg 4801 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat 4861 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg 4921 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca 4981 cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc 5041 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 5101 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc 5161 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 5221 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 5281 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 5341 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 5401 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 5461 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 5521 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa tttt //