LOCUS       V031044                 7632 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031044
VERSION     V031044
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..3779
                     /label="Icam5(NM_008319)"
                     /note="Icam5(NM_008319)"
                     /gene="Icam5"
     CDS             3798..3827
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3846..3869
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(3889..3905)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3916..4538
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(4687..5275)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    5604..5651
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5883..6677)
                     /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(6712..7069)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(7071..7175)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7202..7630
                     /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 gccggggcct tcgccagggc tgcgccgagc gctcctcggc ctctgggctg
     1081 ccctgggcct ggggatccta ggcatctcag cggtcgcgct agaacctttc tgggcggacc
     1141 ttcagccccg cgtggcgctc gtggagcgcg ggggctcgct gtggctcaac tgcagcacta
     1201 actgtccgcg gccggagcgc ggtggcctgg agacctcgct acgccgaaac gggacccaga
     1261 ggggtctgcg ctggctggct cggcagctgg tggacatccg agagcctgag acccagccgg
     1321 tctgcttctt ccgctgcgcg cgccgcacac tccaagcgcg tggactcatc cgaactttcc
     1381 agcgaccgga tcgggtagag ctggtgccgc tcccttcttg gcagccagtg ggcgagaact
     1441 tcaccttgag ctgcagggtc ccgggagcag gaccccgagc gagcctcaca ttgaccctgc
     1501 tgcggggcgg ccaggagctg attcgccgca gtttcgtagg cgagccaccc cgagcgcggg
     1561 gtgcgatgct caccgccagg gtcctggcac gcagggagga ccacagggtc aatttctcat
     1621 gcctcgccga gcttgacctg cggccacacg gcttgggact gtttgcaaat agctcggccc
     1681 ccagacagct ccgtaccttt gccatgcctc cacattcccc gagtcttatt gctccccgag
     1741 tcttagaagt ggactcagaa aggccggtga cttgcacgtt ggatggactg tttcctgccc
     1801 cagaagccgg ggtttacctc tctctgggag atcagaggct caatcctaat gtgaccctcg
     1861 atggggacag ccttgtggcc actgccacag ctacagcaag cgcagaacag gaaggcacca
     1921 aacagctgat gtgcgtcgtg accctcgggg gcgaaagcag ggagacccag gaaaacctga
     1981 ctgtttacag cttcccgact cctcttctga ctttgagtga gccagaagcc cccgagggaa
     2041 aaatggtgac cataagctgc tgggcagggg ctcgagccct tgtcaccctg gagggaattc
     2101 cagctgcggt cccggggcag cccgctgagc tccagttaaa tgttacaaag aacgatgaca
     2161 agcggggctt cttctgcgac gccgccctcg atgtggacgg ggaaactcta agaaaaaacc
     2221 agagctctga gcttcgtgtc ctgtatgcac ctcgcctgga tgacttggat tgtcccagaa
     2281 gctggacgtg gccagagggt ccagagcaga ccctccactg cgaggcccgc ggaaaccctg
     2341 agccctcggt gcactgtgca aggcctgagg gcggggcggt gctagcgctg ggcctattgg
     2401 gtccagtgac ccgtgccctc gcaggcactt accgatgcac agcagtcaat gggcaaggcc
     2461 aggcggtcaa agatgtgacc ctgaccgtgg aatatgcccc agcgctggac agtgtaggct
     2521 gcccagaaca tattacttgg ctggagggaa cggaggcatc gcttagctgt gtggcacacg
     2581 gggtcccacc acctagcgtg agttgtgtgc gctctggaaa ggaggaagtc atggaagggc
     2641 ctctgcgcgt ggcccgggag cacgccggca cttaccgatg cgaagccatc aacgccagag
     2701 gatcagcggc caaaaatgtg gccgtcacgg tggaatatgg tcccagtttt gaggagttgg
     2761 gctgccccag caactggacg tgggtagagg gatctggaaa gctgttttcc tgtgaagttg
     2821 atgggaagcc agaaccacgt gtggagtgcg taggctcgga gggtgcaagc gaagggatag
     2881 tgttgccctt ggtgtcctca aactctggtc ctagaaactc tatgacccct ggtaacctgt
     2941 caccgggcat ttacctctgc aacgccacca accggcacgg ctccacagtc aaaacagtcg
     3001 tcgtgagcgc ggagtcgccg ccacagatgg atgaatccag ttgcccgagt caccagacct
     3061 ggctggaagg agctgaggct accgcgctgg cctgcagtgc caggggccgc ccctctccac
     3121 gggtgcattg ctccagggag ggtgccgcca ggctggagag gctacaggtg tcccgagagg
     3181 atgcggggac ctaccgctgt gtggccacca atgcgcatgg cacggattca cggacggtca
     3241 ctgtgggtgt ggaataccgg cctgtggtgg ctgagctggc agcctcgccc ccaagcgtgc
     3301 ggccgggcgg taacttcact ctgacctgcc gcgcagaggc ttggcctcca gcccagatca
     3361 gctggcgcgc gcccccgggg gctctcaacc tcggcctctc cagcaacaat agcactctga
     3421 gcgtggcggg agccatgggc agccatggcg gcgagtatga gtgtgcagcg accaatgcgc
     3481 atgggcgcca cgcacggcgc atcacagtgc gcgtggccgg tccatggctg tgggtcgctg
     3541 tgggcggtgc ggcagggggc gcggcgctgc tggccgctgg ggccggcctg gccttctacg
     3601 tgcaatccac cgcttgcaag aagggcgagt acaacgtcca ggaggctgag agctcaggcg
     3661 aggccgtgtg tctcaatggc gcgggcggga caccgggtgc agaaggcgga gcagagaccc
     3721 caggcactgc agagtcacct gcagatggcg aggttttcgc tatccagctg acatcttcca
     3781 cgcgtacgcg gccgctcgag cagaaactca tctcagaaga ggatctggca gcaaatgata
     3841 tcctggatta caaggatgac gacgataagg tttaaacggc cggccgcggt catagctgtt
     3901 tcctgaacag atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     3961 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     4021 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     4081 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     4141 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     4201 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     4261 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     4321 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     4381 ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat
     4441 aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc
     4501 tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcggcgagc
     4561 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4621 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4681 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4741 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4801 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4861 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4921 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4981 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5041 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5101 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5161 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5221 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5281 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5341 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5401 ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc
     5461 gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga
     5521 agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc
     5581 ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct
     5641 ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag
     5701 cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca
     5761 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc
     5821 gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc
     5881 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     5941 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     6001 cgggtagcca acgctatgtc ctgatagcga tccgccacac ccagccggcc acagtcgatg
     6061 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     6121 acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc
     6181 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     6241 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     6301 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     6361 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     6421 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     6481 cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     6541 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     6601 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     6661 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat ctttgcaaaa
     6721 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     6781 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     6841 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     6901 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     6961 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     7021 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt
     7081 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7141 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7201 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     7261 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     7321 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     7381 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     7441 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     7501 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     7561 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     7621 aacgcgaatt tt
//
