LOCUS       V030902                 5799 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030902
VERSION     V030902
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..1946
                     /label="Btla(NM_001037719)"
                     /note="Btla(NM_001037719)"
                     /gene="Btla"
     CDS             1965..1994
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2013..2036
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(2056..2072)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    2083..2705
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(2854..3442)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    3771..3818
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(4050..4844)
                     /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(4879..5236)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(5238..5342)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5369..5797
                     /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 gaagacagtg cctgccatgc ttgggactcc tcggttattt agggaattct
     1081 tcatcctcca tctgggcctc tggagcatcc tttgtgagaa agctactaag aggaatgatg
     1141 aagagtgtcc agtgcaactt actattacga ggaattccaa acagtctgcc aggacaggag
     1201 agttatttaa aattcaatgt cctgtgaaat actgtgttca tagacctaat gtgacttggt
     1261 gtaagcacaa tggaacaatc tgtgtacccc ttgaggttag ccctcagcta tacactagtt
     1321 gggaagaaaa tcaatcagtt ccggtttttg ttctccactt taaaccaata catctcagtg
     1381 ataatgggtc gtatagctgt tctacaaact tcaattctca agttattaat agccattcag
     1441 taaccatcca tgtgacagaa aggactcaaa actcttcaga acacccacta ataacagtat
     1501 ctgacatccc agatgccacc aatgcctcag gaccatccac catggaagag aggccaggca
     1561 ggacttggct gctttacacc ttgcttcctt tgggggcatt gcttctgctc cttgcctgtg
     1621 tctgcctgct ctgctttctg aaaaggatcc aagggaaaga aaagaagcct tctgacttgg
     1681 caggaaggga cactaacctg gttgatattc cagccagttc caggacaaat caccaagcac
     1741 tgccatcagg aactggaatt tatgataatg atccctggtc tagcatgcag gatgaatctg
     1801 aattgacaat tagcttgcaa tcagagagaa acaaccaggg cattgtttat gcttctttga
     1861 accattgtgt tattggaagg aatccaagac aggaaaacaa catgcaggag gcacccacag
     1921 aatatgcatc catttgtgtg agaagtacgc gtacgcggcc gctcgagcag aaactcatct
     1981 cagaagagga tctggcagca aatgatatcc tggattacaa ggatgacgac gataaggttt
     2041 aaacggccgg ccgcggtcat agctgtttcc tgaacagatc ccgggtggca tccctgtgac
     2101 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     2161 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     2221 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     2281 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     2341 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     2401 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     2461 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     2521 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttaaa ataactatac
     2581 cagcaggagg acgtccagac acagcatagg ctacctggcc atgcccaacc ggtgggacat
     2641 ttgagttgct tgcttggcac tgtcctctca tgcgttgggt ccactcagta gatgcctgtt
     2701 gaattgggta cgcggccagc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2761 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2821 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2881 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2941 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3001 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3061 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3121 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3181 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3241 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3301 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3361 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3421 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3481 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3541 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaacc
     3601 tgaggctatg gcagggcctg ccgccccgac gttggctgcg agccctgggc cttcacccga
     3661 acttgggggg tggggtgggg aaaaggaaga aacgcgggcg tattggcccc aatggggtct
     3721 cggtggggta tcgacagagt gccagccctg ggaccgaacc ccgcgtttat gaacaaacga
     3781 cccaacaccg tgcgttttat tctgtctttt tattgccgtc atagcgcggg ttccttccgg
     3841 tattgtctcc ttccgtgttt cagttagcct ccccctaggg tgggcgaaga actccagcat
     3901 gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa
     3961 cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg
     4021 gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag
     4081 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     4141 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcgatcc
     4201 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     4261 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgctcgcc
     4321 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     4381 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     4441 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     4501 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     4561 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     4621 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca
     4681 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     4741 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     4801 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     4861 cctgtctctt gatcgatctt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     4921 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     4981 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     5041 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5101 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
     5161 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg
     5221 gttctttccg cctcaggact cttccttttt caatattatt gaagcattta tcagggttat
     5281 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5341 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg
     5401 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5461 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5521 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5581 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5641 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5701 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5761 aaaaatgagc tgatttaaca aaaatttaac gcgaatttt
//
