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&nbsp;(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
//
