LOCUS       V020523                 6318 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020523
VERSION     V020523
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             1005..2480
                     /label="Thra(NM_001313983)"
                     /note="Thra(NM_001313983)"
                     /gene="Thra"
     CDS             2484..2513
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2532..2555
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(2575..2591)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    2602..3224
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3373..3961)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4290..4337
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(4569..5363)
                     /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(5398..5755)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(5757..5861)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5888..6316
                     /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 tggatccagc caccatggaa cagaagccaa
     1021 gcaaggtgga gtgtgggtca gacccagagg agaacagtgc caggtcacca gatggaaagc
     1081 gaaaaagaaa gaacggccaa tgtcccctga aaagcagcat gtcagggtat atccctagtt
     1141 acctggacaa agacgagcag tgtgtcgtgt gtggggacaa ggccaccggt tatcactacc
     1201 gctgtatcac ttgtgagggc tgcaagggct tctttcgccg cacaatccag aagaatctcc
     1261 atcccaccta ttcctgcaag tatgacagct gctgtgtcat cgacaagatc acccggaatc
     1321 agtgccagct gtgccgcttc aagaagtgca ttgctgtggg catggccatg gacttggttc
     1381 tagatgattc gaagcgggtg gccaaacgca agctgattga gcagaaccgg gagaggaggc
     1441 gaaaggagga gatgattcgc tcactgcagc agcgaccaga gcccactcct gaagagtggg
     1501 acctgataca tgttgctaca gaggcccatc gcagcactaa cgcccagggc agccattgga
     1561 aacagaggcg aaaattcctg ccggatgaca ttggccagtc acctattgtc tccatgccgg
     1621 acggagacaa ggtagaccta gaggccttca gcgagtttac caagatcatc accccggcca
     1681 tcacgcgcgt ggtggacttt gccaaaaaac tgcccatgtt ctccgagctg ccttgcgaag
     1741 accagatcat cctcctgaag ggctgctgca tggagatcat gtccctgcgg gcagctgtcc
     1801 gctacgaccc tgagagtgac accctgaccc tgagtgggga gatggcggtt aagcgggaac
     1861 agctcaagaa tggtggcttg ggtgtggtct ctgacgccat ctttgaactg ggcaagtcac
     1921 tctctgcctt taacctggat gacacggaag tggctctgct gcaggctgtg ctgctaatgt
     1981 caacagaccg ctccggcctg ctgtgcgtgg acaagatcga gaagagtcag gaggcctacc
     2041 tgctggcgtt tgagcactac gtcaaccacc gcaaacacaa cattccgcac ttctggccca
     2101 agctgctgat gaaggagaga gaagtgcaga gttcgattct gtacaagggg gcagcggcag
     2161 aaggccggcc gggcgggtca ctgggcgtcc acccggaagg acagcagctt ctcggaatgc
     2221 atgttgttca gggtccgcag gtccggcagc ttgagcagca gcttggtgaa gcgggaagtc
     2281 tccgagggcc ggttcttcag caccagagcc cgaagagccc gcagcagcgt ctcctggagc
     2341 tgctccaccg aagcggaatt ctccattccc gagcggtctg tggggaagac gacagcagtg
     2401 aggcgagctc cctgagctcc tcctctgaca cggaggacac ggaggtctgc gaggaccagg
     2461 caggcaaggc agcctctccc ctcgagcaga aactcatctc agaagaggat ctggcagcaa
     2521 atgatatcct ggattacaag gatgacgacg ataaggttta aacggccggc cgcggtcata
     2581 gctgtttcct gaacagatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     2641 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     2701 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     2761 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     2821 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     2881 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     2941 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     3001 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     3061 gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca
     3121 cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact
     3181 gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagcg
     3241 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3301 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3361 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3421 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3481 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3541 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3601 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3661 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3721 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3781 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3841 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3901 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3961 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4021 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4081 atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc
     4141 cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga
     4201 aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg
     4261 ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt
     4321 ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc
     4381 agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat
     4441 catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt
     4501 ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag
     4561 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     4621 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     4681 atatcacggg tagccaacgc tatgtcctga tagcgatccg ccacacccag ccggccacag
     4741 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     4801 tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg
     4861 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
     4921 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
     4981 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     5041 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     5101 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     5161 gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca
     5221 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     5281 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     5341 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcgatcttt
     5401 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     5461 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     5521 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     5581 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     5641 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     5701 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc
     5761 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5821 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5881 ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5941 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6001 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6061 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     6121 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6181 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6241 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6301 aaatttaacg cgaatttt
//
