LOCUS       V031029                 5979 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031029
VERSION     V031029
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..2126
                     /label="Usp46(NM_177561)"
                     /note="Usp46(NM_177561)"
                     /gene="Usp46"
     CDS             2145..2174
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2193..2216
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(2236..2252)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    2263..2885
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3034..3622)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    3951..3998
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(4230..5024)
                     /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(5059..5416)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(5418..5522)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5549..5977
                     /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 gactgtccga aacatcgcct ccatctgtaa tatgggcacc aatgcctctg
     1081 ctctggaaaa agacattgga ccagagcagt ttccgatcaa tgaacactac ttcggattgg
     1141 tcaattttgg aaacacatgc tactgtaact ccgtgcttca agcgctgtac ttctgccggc
     1201 cgttccggga gaatgtgttg gcatacaagg cccagcaaaa aaagaaggaa aacctgttga
     1261 catgcctggc agaccttttc cacagcattg ccacgcagaa gaagaaggtt ggcgtcatcc
     1321 caccaaagaa gttcatttca aggctgagga aagagaacga tctctttgat aactacatgc
     1381 agcaggatgc tcacgagttc ttaaattatt tgctaaacac tattgcggac atcctgcagg
     1441 aagagaagaa acaggaaaaa caaaacggga agttaaaaaa cggcaacatg aacgaaccag
     1501 ccgagaacaa taagccagaa ctcacctggg tccacgagat ttttcaggga acactcacta
     1561 acgaaactcg atgcttgaac tgtgaaactg tgagtagcaa agatgaagat tttcttgacc
     1621 tctctgtcga tgtggaacag aacacctcca tcacccactg tctgagagac ttcagcaaca
     1681 cagaaacgct gtgtagtgaa caaaaatatt attgtgaaac atgttgcagc aaacaggagg
     1741 ctcagaagag gatgagggta aaaaagctgc ccatgatttt ggccctgcac ctcaagaggt
     1801 tcaagtacat ggaacagctc cacaggtaca ccaagctgtc ttaccgggtg gtcttccctc
     1861 tggaactccg gctcttcaac acctccagcg atgctgtgaa cctggaccgc atgtatgacc
     1921 tggttgctgt ggtcgttcac tgtggaagtg gtcctaaccg tgggcattac atcaccatcg
     1981 taaaaagtca tggcttctgg ctgttgtttg atgacgacat tgtagagaaa atagacgctc
     2041 aagccattga ggaattctat ggtctaacgt cagatatatc aaaaaattca gaatctggat
     2101 atattttatt ctatcaatca agagaaacgc gtacgcggcc gctcgagcag aaactcatct
     2161 cagaagagga tctggcagca aatgatatcc tggattacaa ggatgacgac gataaggttt
     2221 aaacggccgg ccgcggtcat agctgtttcc tgaacagatc ccgggtggca tccctgtgac
     2281 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     2341 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     2401 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     2461 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     2521 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     2581 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     2641 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     2701 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttaaa ataactatac
     2761 cagcaggagg acgtccagac acagcatagg ctacctggcc atgcccaacc ggtgggacat
     2821 ttgagttgct tgcttggcac tgtcctctca tgcgttgggt ccactcagta gatgcctgtt
     2881 gaattgggta cgcggccagc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2941 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3001 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3061 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3121 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3181 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3241 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3301 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3361 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3421 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3481 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3541 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3601 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3661 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3721 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaacc
     3781 tgaggctatg gcagggcctg ccgccccgac gttggctgcg agccctgggc cttcacccga
     3841 acttgggggg tggggtgggg aaaaggaaga aacgcgggcg tattggcccc aatggggtct
     3901 cggtggggta tcgacagagt gccagccctg ggaccgaacc ccgcgtttat gaacaaacga
     3961 cccaacaccg tgcgttttat tctgtctttt tattgccgtc atagcgcggg ttccttccgg
     4021 tattgtctcc ttccgtgttt cagttagcct ccccctaggg tgggcgaaga actccagcat
     4081 gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa
     4141 cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg
     4201 gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag
     4261 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     4321 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcgatcc
     4381 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     4441 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgctcgcc
     4501 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     4561 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     4621 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     4681 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     4741 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     4801 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca
     4861 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     4921 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     4981 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     5041 cctgtctctt gatcgatctt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     5101 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     5161 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     5221 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5281 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
     5341 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg
     5401 gttctttccg cctcaggact cttccttttt caatattatt gaagcattta tcagggttat
     5461 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5521 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg
     5581 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5641 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5701 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5761 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5821 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5881 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5941 aaaaatgagc tgatttaaca aaaatttaac gcgaatttt
//
