LOCUS       V031251                 8301 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031251
VERSION     V031251
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..4448
                     /label="Trim33(NM_053170)"
                     /note="Trim33(NM_053170)"
                     /gene="Trim33"
     CDS             4467..4496
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             4515..4538
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(4558..4574)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    4585..5207
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(5356..5944)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    6273..6320
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(6552..7346)
                     /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(7381..7738)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(7740..7844)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7871..8299
                     /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 ggcggaaaac aaaggcggcg gtgaggcaga gagtggcggc gggggcagcg
     1081 gtagcgcgcc ggtaaccgcc ggggccgccg ggcccacggc gcaggaggcg gagccgcctc
     1141 tcgccgcggt gctggtggag gaggaggagg aggaaggcgg cagggctggc gcggagggcg
     1201 gtgcggctgg gcccgacgac gggggggtgg ccgcggcctc ctcaagctcg gccccggccg
     1261 cctcggtccc cgcggcctca gtgggctccg cggtccccgg aggcgcggca tcgacgccgg
     1321 ctccagctgc tgccccagcc ccggccccgg ccccggcccc ggccccggcc cccgctcccg
     1381 ccccggctcc tggttcctcg tcggggccac ctctcggacc accagcctcg cttctggaca
     1441 cctgcgcggt gtgtcagcag agcttgcaga gccggcgcga ggcggagccc aagctgctcc
     1501 cctgcctgca ctccttctgc ctgcgctgtc tgccggagcc ggagcgccag ctcagcgtgc
     1561 ccatccctgg gggcagcaac ggtgacgtcc agcaagttgg tgtaatacgg tgtccagtat
     1621 gtcgtcaaga atgtagacag atagatcttg tggataatta ttttgtgaaa gacacatctg
     1681 aagctcctag cagctctgat gagaaatcag aacaggtatg tactagttgt gaagataatg
     1741 caagtgcagt tggtttttgt gtagaatgtg gagagtggct gtgtaagaca tgcattgaag
     1801 cacatcaaag agtaaaattc actaaagacc acctgatcag gaaaaaagaa gatgtctcag
     1861 agtctgttgg aacatctggt cagcgcccag tgttttgtcc tgtacataaa caagagcagt
     1921 tgaaactatt ctgtgaaacg tgtgatagat tgacgtgtag agactgtcaa cttttggagc
     1981 acaaagaaca taggtatcag tttttagaag aggcttttca aaatcagaaa ggtgcaattg
     2041 aaaatctgtt ggctaagctt cttgaaaaga agaattatgt tcattttgca gctacacaag
     2101 tgcagaatag gattaaagaa gtaaatgaga ctaacaaacg tgtagaacag gaaattaaag
     2161 tggccatctt cacccttatc aatgaaatta ataagaaagg aaaatctctc ttgcaacagc
     2221 tagagaatgt tacaaaagaa agacagatga agttactaca gcagcagaat gatatcacag
     2281 gcctctcccg gcaggtgaag catgttatga actttacaaa ctgggccatt gcaagtggca
     2341 gcagcacggc cctactttat agcaagcgtc tgattacttt tcagttacgt catattttga
     2401 aagcccgatg tgatcctgtt cctgctgcca atggagcaat tcgtttccat tgtgatccca
     2461 ccttctgggc caaaaatgta gtgaatttag gtaatctagt aatagagagt aagccagctc
     2521 ctggttatac tcctaatgtt gtagttgggc aagttcctcc agggacaaac cacattagta
     2581 aaacccctgg acagattaat ttagcacagc ttcgactcca gcacatgcag caacaagtgt
     2641 atgcacagaa acatcagcag ttacaacaga tgagactgca gcaaccacca gctcctatac
     2701 caacaacaac agcaacaacc cagcagcatc ctaggcaagc agccccacag atgttacaac
     2761 aacagcctcc aagattgatc agtgtgcaaa caatgcaaag aggcaacatg aactgtggag
     2821 catttcaagc ccatcaaatg agactggctc agaatgctgc cagaatacca gggatcccca
     2881 ggcacagtgc ccctcagtat tccatgatgc agccacatct ccaaagacaa cattcaaatc
     2941 caggacatgc tggaccgttt cctgttgtat cagcacacaa cccaatcaac ccaacaagtc
     3001 ccaccacagc aactatggca aatgcaaatc gaggtcccac cagcccgtct gttacagcaa
     3061 tagaattaat accctcagtc accaatccag aaaaccttcc atcgttgcca gatatcccac
     3121 ctatacagtt ggaagatgct ggctcaagta gtttagataa tctactgagt agatatatct
     3181 caggcagtca tctaccccct cagcctacaa gtaccatgaa tccttcccca ggaccctctg
     3241 cgttatcacc tggatcttca ggcttatcca attctcacac acctgtgaga ccccctagca
     3301 cgtctagtac tggcagtcga ggcagctgcg gatcatcagg aagaactgct gagaaaagtg
     3361 ctcatagttt caagtctgat caggtgaagg tcaagcagga acctgggact gaagaagaga
     3421 tatgcagctt ttcaggagct gtgaaacaag agaagacaga ggatggcagg aggagtgcct
     3481 gtatgttgag cagtcctgag agtagtttga caccacctct ttcaactaac cttcacctag
     3541 agagtgagct ggatacatta acaggcctgg agaaccacgt gaagactgaa cctacagata
     3601 tcagtgaaag ctgcaaacag tcaggactca gtaacctagt caatggaaag tctccaattc
     3661 gaaacctcat gcacaggtcg gcaaggattg gaggggatgg caatagcaaa gatgatgacc
     3721 caaatgaaga ctggtgtgcc gtctgccaga acggagggga tcttttgtgt tgtgagaaat
     3781 gtccgaaggt ctttcatctt acctgtcatg ttccaacact tctcagcttt ccaagtggag
     3841 actggatatg cacattttgc agagatattg ggaagccaga agttgaatat gattgtgata
     3901 atatgcagca tagtaagaag gggaaaactg cacaaggctt aagccctgtg gaccaaagga
     3961 aatgtgaacg tcttctgctt tacctctatt gtcacgaatt aagtattgaa ttccaggagc
     4021 ctgttcctgt ttcgatacca aactactata aaattataaa gaaaccgatg gatttatcaa
     4081 cggtgaagaa gaagcttcag aaaaaacatt cccagcatta ccaaatccca gatgactttg
     4141 tggctgatgt ccgtttgatc ttcaagaact gtgaaaggtt taatgaaatg atgaaagttg
     4201 ttcaagttta tgcagacaca caagagatta atttgaaggg tgattcagaa gtagctaagg
     4261 cagggaaagc agttgcattg tactttgaag ataaactctc agagatctac tcagacagga
     4321 ccttcactcc tttgccagag tttgagcagg acgaagatga tggtgaagta acagaagact
     4381 ctgacgaaga ttttatacag cctcgtagaa aacgcctaaa gtcagacgag agaccagtac
     4441 atataaagac gcgtacgcgg ccgctcgagc agaaactcat ctcagaagag gatctggcag
     4501 caaatgatat cctggattac aaggatgacg acgataaggt ttaaacggcc ggccgcggtc
     4561 atagctgttt cctgaacaga tcccgggtgg catccctgtg acccctcccc agtgcctctc
     4621 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     4681 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     4741 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     4801 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     4861 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     4921 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     4981 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     5041 actgctccct tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag
     5101 acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc
     5161 actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca
     5221 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5281 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5341 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5401 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5461 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5521 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5581 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5641 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5701 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5761 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5821 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5881 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5941 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6001 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6061 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc
     6121 tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg
     6181 ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga
     6241 gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt
     6301 attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt
     6361 ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     6421 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     6481 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     6541 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     6601 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     6661 gcaatatcac gggtagccaa cgctatgtcc tgatagcgat ccgccacacc cagccggcca
     6721 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     6781 ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt
     6841 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     6901 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     6961 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     7021 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     7081 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     7141 cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg
     7201 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     7261 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     7321 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc
     7381 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     7441 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     7501 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     7561 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     7621 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     7681 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagga
     7741 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7801 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7861 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     7921 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     7981 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     8041 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     8101 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     8161 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     8221 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     8281 caaaaattta acgcgaattt t
//
