LOCUS V030955 5967 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030955 VERSION V030955 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..2114 /label="Msi1(NM_008629)" /note="Msi1(NM_008629)" /gene="Msi1" CDS 2133..2162 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 2181..2204 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" primer_bind complement(2224..2240) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 2251..2873 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(3022..3610) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 3939..3986 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(4218..5012) /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(5047..5404) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(5406..5510) /gene="bla" /label="AmpR promoter" rep_origin 5537..5965 /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 ggagactgac gcgccccagc ccggcctcgc ctccccggac tcgccgcacg 1081 acccctgcaa gatgttcatc ggaggactca gttggcagac cacgcaggaa gggctgcgcg 1141 aatacttcgg ccagttcggg gaggtgaaag agtgtctggt gatgcgggac cccctgacca 1201 aaagatccag gggtttcggc ttcgtcactt tcatggacca ggcgggggtg gataaagtgc 1261 tggcgcaatc gcggcacgag ctcgactcca aaacaattga ccccaaggtg gcctttcctc 1321 gaagagcaca gcctaagatg gtcactcgga cgaagaagat cttcgtgggg gggctgtctg 1381 tgaacaccac ggtggaagat gtgaaacact atttcgagca gttcggaaag gtggatgatg 1441 ccatgctgat gttcgacaaa accaccaaca ggcacagagg gtttggattt gtcacgtttg 1501 agagcgagga catcgtagag aaagtttgtg agatccactt ccatgaaatc aacaacaaaa 1561 tggtggaatg caagaaagcc cagccaaagg aggtgatgtc cccgacaggc tcagcccggg 1621 gcaggtctcg ggtcatgccc tacggaatgg atgccttcat gctgggtatt gggatgctgg 1681 gttacccagg gttccaagcc acgacctacg ccagccggag ttacacaggc cttgcccctg 1741 gttacaccta ccagttcccc gaattccgtg tagagcggag ccctctcccg agcgccccag 1801 tcctccccga gctcacagct atccctctca cggcttatgg gcccatggcg gcggcagcgg 1861 cggcggcagc tgtagttcga gggacaggct ctcacccctg gacgatggct ccccctccag 1921 gttccactcc cagccgcaca gggggcttcc tagggaccac aagccccggc cccatggctg 1981 agctctacgg ggcagccaac caggactccg gggtcagcag ttacatcagc gccgccagcc 2041 ccgcccccag cactggtttc ggccacagtc ttgggggtcc cttgattgcc acagccttca 2101 ccaatgggta ccacacgcgt acgcggccgc tcgagcagaa actcatctca gaagaggatc 2161 tggcagcaaa tgatatcctg gattacaagg atgacgacga taaggtttaa acggccggcc 2221 gcggtcatag ctgtttcctg aacagatccc gggtggcatc cctgtgaccc ctccccagtg 2281 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta 2341 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg 2401 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa 2461 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa 2521 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc 2581 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc 2641 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg 2701 tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac 2761 gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg 2821 cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg 2881 cggccagcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 2941 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 3001 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 3061 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 3121 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 3181 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 3241 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 3301 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 3361 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 3421 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 3481 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 3541 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 3601 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 3661 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 3721 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc 3781 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg 3841 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc 3901 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg 3961 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt 4021 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg 4081 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa 4141 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc 4201 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 4261 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 4321 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcgatccgc cacacccagc 4381 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 4441 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg 4501 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 4561 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 4621 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 4681 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 4741 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 4801 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg 4861 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 4921 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 4981 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga 5041 tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc 5101 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg 5161 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat 5221 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga 5281 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg 5341 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc 5401 tcaggactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 5461 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 5521 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 5581 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 5641 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 5701 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 5761 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 5821 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 5881 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 5941 atttaacaaa aatttaacgc gaatttt //