LOCUS       V031113                10137 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031113
VERSION     V031113
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..6284
                     /label="Itgb4(NM_001405126)"
                     /note="Itgb4(NM_001405126)"
                     /gene="Itgb4"
     CDS             6303..6332
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             6351..6374
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(6394..6410)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    6421..7043
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(7192..7780)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    8109..8156
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(8388..9182)
                     /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(9217..9574)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(9576..9680)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      9707..10135
                     /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 ggcagggccc tgttgcagcc catgggtgaa gctgctgctg ctggcagcaa
     1081 tgctgagtgc cagcctccct ggagacctgg ccaaccgctg caagaaggct caggtgaaga
     1141 gctgtaccga gtgcatccgg gtggacaaga gctgtgccta ctgcacagac gagctgttca
     1201 aggagaggcg ctgcaacacc caggcggagc ttctggctgc aggctgcagg ggagagagca
     1261 tcctggtcat ggagagcagc cttgaaatca cagagaacac ccagatcgac accagcctgc
     1321 accgcagcca ggtatctccc caaggcctgc aagtccggct gcggccgggt gaggagcgca
     1381 gctttgtgtt ccaggtgttt gagcccctgg agagccccgt ggatctgtat atcctcatgg
     1441 acttctccaa ctccatgtct gacgatctgg acaacctcaa gcagatgggg cagaacctgg
     1501 ccaagatcct gcgccagctc accagcgact acaccattgg atttggaaag tttgtggaca
     1561 aagtcagcgt cccacagaca gacatgaggc ccgagaaact gaaggagccc tggcccaaca
     1621 gtgatccccc gttctccttc aagaacgtta tcagcttaac ggagaatgtg gaagaattct
     1681 ggaacaaact gcaaggagaa cgcatctcag gcaacctgga cgctcctgaa gggggctttg
     1741 atgccatcct gcagacagct gtgtgcacaa gggacattgg ctggagggct gacagcaccc
     1801 acctgctggt gttctccacc gagtctgcct tccactacga ggctgatggt gccaacgtgc
     1861 tggccggcat catgaaccgc aatgatgaga aatgccacct ggacgcctcg ggcgcctaca
     1921 cccaatacaa gacacaggac tacccatcag tgcccacgct ggttcgcctg cttgccaagc
     1981 ataacatcat ccccatcttt gctgtcacca actactctta cagctactat gagaagctcc
     2041 ataagtattt ccccgtctcc tctctgggcg tcctgcagga ggattcatcc aacatcgtgg
     2101 agctgctgga ggaggccttc tatcgaattc gctccaacct ggacatccgg gctctggaca
     2161 gccccagagg cctgagaaca gaggtcacct ccgatactct ccagaagacg gagactgggt
     2221 cctttcacat caagcggggg gaagtgggca catacaatgt gcatctccgg gcagtggagg
     2281 acatagatgg gacacatgtg tgccagctgg ctaaagaaga ccaagggggc aacatccacc
     2341 tgaaaccctc cttctctgat ggcctccgga tggacgcgag tgtgatctgt gacgtgtgcc
     2401 cctgtgagct gcaaaaggaa gttcgatcag ctcgctgtca cttcagagga gacttcatgt
     2461 gtggacactg tgtgtgcaat gagggctgga gtggcaaaac ctgcaactgc tccaccggct
     2521 ctctgagtga cacacagccc tgcctgcgtg agggtgagga caaaccgtgc tcgggccacg
     2581 gcgagtgcca gtgcggacgc tgtgtgtgct atggtgaagg ccgctacgag ggtcacttct
     2641 gcgagtatga caacttccag tgtccccgga cctctggatt cctgtgcaat gaccggggtc
     2701 gctgttctat gggagagtgt gtgtgtgagc ctggttggac aggccgcagc tgtgactgtc
     2761 ccctcagcaa tgccacctgc atcgatagca acgggggcat ctgcaacggc cgaggctact
     2821 gtgagtgtgg ccgttgtcac tgcaaccagc agtcgctcta cacggacacc acctgtgaga
     2881 tcaactactc tgcgatacgc ctgggtctct gtgaggatct ccgctcctgc gtacagtgcc
     2941 aggcctgggg caccggggag aagaaagggc gcgcgtgtga cgattgcccc tttaaagtca
     3001 agatggtaga cgagcttaag aaagaagagg tggtggagta ctgctccttc cgggatgagg
     3061 atgacgactg cacttacagc tacaacgtgg agggcgatgg cagccctggg cccaacagca
     3121 cagtcctggt ccacaaaaag aaagactgcc ctcccggctc cttctggtgg ctcatccccc
     3181 tgctcatctt cctcctgttg ctcctggcgt tgcttctgct gctctgctgg aaatactgtg
     3241 cctgctgcaa agcctgcctg gggcttcttc cttgctgcaa ccgaggtcac atggtgggct
     3301 ttaaggaaga tcactatatg cttcgggaga acctgatggc ctctgaccac ctggacacgc
     3361 ccatgctacg aagcgggaac ctcaagggac gagacacagt ccgctggaag atcaccaaca
     3421 atgtgcagcg ccctggcttt gccacccatg ccgccagcac cagccccacg gagctcgtac
     3481 cctacgggct gtccctgcgc cttggccgcc tctgcactga gaaccttatg aagccgggca
     3541 cccgagagtg tgaccagcta cgccaggagg tggaggaaaa tctgaatgag gtgtatagac
     3601 aggtcagcgg cgcacacaag ctccagcaga cgaagttccg acagcagccc aacaccggga
     3661 aaaagcaaga ccacaccatt gtggacacag tgttgctggc gccccgctcc gccaagcaga
     3721 tgctgctgaa gctgacagag aagcaggtgg agcaggggtc cttccatgaa ctgaaggtgg
     3781 cccctggcta ctacactgtc acggcagagc aggatgcccg gggcatggtg gagttccagg
     3841 agggcgtgga gctggtggat gtgcgagtgc ccctcttcat ccggcctgag gatgatgatg
     3901 agaagcagct gctggtggag gccattgatg tccctgtggg cactgccacc cttggtcgcc
     3961 gtctggtaaa catcaccatt atcaaggaac aagctagtgg ggtagtgtcc ttcgagcagc
     4021 ctgaatactc ggtgagtcgt ggagaccagg tggcccgcat ccctgtcatc cggcacatcc
     4081 tggacaatgg caagtcccag gtctcctata gcacacagga taatacagca cacggacacc
     4141 gggattatgt ccctgtggag ggagagctgc tgttccatcc tggggagacc tggaaggagt
     4201 tgcaggtgaa gctactggag ctgcaggagg tcgactccct cctgcgtggc cgccaggtcc
     4261 gccgcttcca agtccaactc agcaacccca agttcggagc ccgcctgggc cagcccagca
     4321 caaccaccgt tattctcggt gagcacgatg aaacggacag gagtctcata aatcaaacac
     4381 tttcatcgcc tccgccaccc catggagacc tgggcgcccc acagaacccc aatgccaagg
     4441 ctgccgggtc caggaagatc catttcaact ggctgccccc tcctggcaag ccaatggggt
     4501 acagggtgaa gtactggatc cagggcgact ctgagtctga agcccacctt ctagacagca
     4561 aggtgccctc agtggaactc accaacctgt atccctattg cgactacgaa atgaaggtgt
     4621 gtgcctatgg ggcacagggt gaggggccct atagctcact ggtgtcctgc cgcacccacc
     4681 aggaagtacc cagcgagcca gggaggctgg ctttcaatgt agtctcttct acggtgacgc
     4741 agctgagctg ggcagagcca gctgagacca atggcgagat cacagcctac gaggtctgct
     4801 atggactggt caatgaggac aaccgaccca ttggacctat gaagaaggtg ctcgtggaca
     4861 accccaagaa ccggatgctg ctcattgaga atctgcgaga atcccagcca taccgataca
     4921 cggttaaggc gcgcaatggg gcaggatggg gacccgagag agaggctatc atcaacctgg
     4981 ctacacagcc caagcggccc atgtccatcc ctatcatccc agacatcccc atagtggacg
     5041 cccagggtgg agaagactac gaaaacttcc ttatgtacag tgatgacgtc ctgcggtccc
     5101 cagccagcag ccagaggccc agcgtttctg atgacactga gcacctggtg aatggccgga
     5161 tggactttgc ctatccaggc agcgccaact ccctgcacag aatgactgca gccaatgtgg
     5221 cctatggcac gcatctgagc ccacacctgt cccaccgagt gctgagcacg tcctccaccc
     5281 ttacccggga ctaccactct ctgacacgca cagagcactc ccactcaggc acacttccca
     5341 gggactactc caccctcact tccctttcct cccaagattc ccgtggggct gtgggtgtgc
     5401 ctgacacacc cactcggctg gtgttctctg ccctggggcc cacgtctttg aaggtgagct
     5461 ggcaggagcc acagtgtgat cggatgctgc tgggctacag tgtggaatac cagctactaa
     5521 acggcggcga gatgcatcgg ctcaacatcc ctaaccctgg ccaaacctcg gtggtggtag
     5581 aggatctcct gcctaaccac tcctatgtgt tccgggtacg ggcccagagc caggagggct
     5641 ggggccgaga gcgagagggt gtcatcacca tcgagtccca ggtgcacccg cagagccctc
     5701 tctgccccct gccaggctca gccttcactc tgagcacccc cagcgccccg ggaccgctgg
     5761 tgttcactgc cctaagccca gactccctgc agctcagctg ggagcggccg aggaggccca
     5821 atggagatat ccttggctac ctggtgacct gtgagatggc ccaaggagga gcaccagcca
     5881 ggaccttccg ggtggacgga gacaaccctg agagccggtt gactgtacct ggcctcagtg
     5941 agaacgttcc ttacaagttc aaggttcagg ccaggacgac cgagggcttc gggccagagc
     6001 gtgagggtat catcaccatc gagtctcagg ttggaggccc cttcccacag ctgggcagcc
     6061 attctgggct cttccagaac ccagtgcaaa gcgagttcag cagcgtgacc agcacgcaca
     6121 gcaccacgac tgagcccttc ctcatggatg gtctaaccct ggggacccag cgcctggaag
     6181 caggaggctc cctcacccgg catgtgaccc aggaattcgt gacccggacc ttaacggcca
     6241 gtggctctct cagcactcat atggaccaac agttcttcca aaccacgcgt acgcggccgc
     6301 tcgagcagaa actcatctca gaagaggatc tggcagcaaa tgatatcctg gattacaagg
     6361 atgacgacga taaggtttaa acggccggcc gcggtcatag ctgtttcctg aacagatccc
     6421 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     6481 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     6541 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     6601 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     6661 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     6721 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     6781 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     6841 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     6901 attttaaaat aactatacca gcaggaggac gtccagacac agcataggct acctggccat
     6961 gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc
     7021 actcagtaga tgcctgttga attgggtacg cggccagcgg cgagcggtat cagctcactc
     7081 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7141 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7201 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7261 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7321 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7381 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7441 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7501 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7561 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7621 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7681 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7741 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7801 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7861 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7921 aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag
     7981 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
     8041 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     8101 gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat
     8161 agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg
     8221 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     8281 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     8341 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     8401 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     8461 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     8521 atgtcctgat agcgatccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     8581 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     8641 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     8701 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     8761 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     8821 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     8881 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     8941 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     9001 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     9061 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     9121 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     9181 atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa
     9241 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     9301 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     9361 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     9421 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     9481 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     9541 acacacattc cacagctggt tctttccgcc tcaggactct tcctttttca atattattga
     9601 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9661 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     9721 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     9781 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     9841 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     9901 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     9961 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
    10021 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
    10081 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaatttt
//
