LOCUS       V031198                 7767 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031198
VERSION     V031198
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..3914
                     /label="Usp4(NM_011678)"
                     /note="Usp4(NM_011678)"
                     /gene="Usp4"
     CDS             3933..3962
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3981..4004
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(4024..4040)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    4051..4673
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(4822..5410)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    5739..5786
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(6018..6812)
                     /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(6847..7204)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(7206..7310)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7337..7765
                     /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 ggcggaaggc cggggcagcc gtgagcgacc ggatgtggag acccagaaga
     1081 cggagctcgg agccttgatg gggaccacgc tccaacgtgg ggcgcagtgg tatcttattg
     1141 acagccggtg gttcaagcag tggaagaagt atgtgggctt tgacagctgg gacatgtaca
     1201 atgtcgggga gcataacctg tttcctggac ctattgacaa ctccggactc ttctcagatc
     1261 ctgagagtca gaccttgaag gagcacttaa tcgatgagct ggactatgtg ctggtcccag
     1321 ccgaagcctg gaataaattg ctgaattggt atggctgtgt ggagggccag cagcctattg
     1381 tcagaaaagt tgtggagcat ggcctgtttg tcaagcactg caaagtggaa gtgtatttgc
     1441 tggagctgaa gctctgtgag aacagtgacc ccaccaatgt gctaagttgc cattttagca
     1501 aagcagacac cattgcaact attgagaagg agatgaggaa gctcttcaac atccctgcag
     1561 aacgtgaaac acggctttgg aacaaataca tgagcaacac ctatgagcag ttgagcaagc
     1621 tagacaacac tatccaggat gctgggctgt accagggtca ggtgctagta attgagcccc
     1681 aaaatgaaga tggcacatgg ccccggcaga gcctgcagtc aaaatcaagc actgcaccta
     1741 gcagaaattt cactacctct tcaaaaccat ccgcaagtcc ctattgctca gtgtctgcct
     1801 ctctcattgc aaatggtgat agcactaaca gctctgggat gcacagctcc ggtgtcagca
     1861 ggggtggatc tggcttctct gcttcgtata attgccagga gcccccatca cctcatatac
     1921 agcctggcct ctgtggactt ggaaacctgg ggaacacttg cttcatgaac tctgctctgc
     1981 agtgcctgag caacactgcc ccactgactg agtactttct caaagatgag tatgaggccg
     2041 agatcaaccg agacaaccct ctggggatga aaggggagat tgcagaggcc tatgcagagc
     2101 tcatcaagca gatgtggtct ggaagggaca ctcacgtggc accacggatg ttcaagacgc
     2161 aagtgggacg ttttgcccct cagttttctg gctaccaaca gcaagactcc caggagctgc
     2221 tagcctttat tctagacgga ctgcacgagg acctgaaccg cgtaaagaag aagccttacc
     2281 tggagcccaa ggacgccaat gggcgaccag atgcggtggt agcaaaggaa gcctgggaaa
     2341 accacaggct gaggaatgat tctgtgattg tggatacttt ccatggcctt ttcaaatcga
     2401 ctttggtttg cccagaatgt gctaaagttt ctgtgacctt tgacccattt tgctatctaa
     2461 ctctcccact gcctttgaag aaggatcgga ttatggaggt cttcctggtt cctgctgacc
     2521 ctcagtgcag acctatccag taccgtgtga ctgtgccatt gatgggggcc atttctgacc
     2581 tgtgtgaagc actctccaag ctgtctggca ttgctgcaga aaatatggtg gtcactgatg
     2641 tatataatca ccgcttccac aaaattttcc aaatggatga aggtttaagc cacatcacgc
     2701 ctcgagatga catttttgtg tatgaggtct gcaacacgtc catggatggc tcagagtgta
     2761 tcactcttcc agtctacttc agagagaaga agtccaggcc gtcgagtgct tcctcggggg
     2821 ctgtgctcta tggacagccc cttcttgtct ctgtccctaa gcataagcta accctagagt
     2881 ctttgtacca ggctgtttgt gatcgtatca gccgctacat aaaacagcct ttgcctgatg
     2941 agtttctcag ctcaccctta gagcctgggg cctgcaatgg ctctaggagt agctatgaag
     3001 gagatgaaga ggaagaaatg gatcatcaag aggaaggaaa agagcagctt tccgaagtgg
     3061 aaggcagtgg tgaggacgat cagggagatg accatagcga gagcgcccaa aaggtgaaag
     3121 gccagccgag gcacaagagg ctctttacct tcagcctcgt gaactcctgt ggaactgctg
     3181 atatcaattc actggcaact gacgggaaac tcctcaaact caactctcga tccacactgg
     3241 ccattgactg ggacagtgag acccgaagcc tttactttga tgagcaagaa tctgaggcct
     3301 gtgagaagca cctgagcatg tcacagccgc agaagaagaa gaaggcggca gtagccctgc
     3361 gagagtgcat cgagctcttc accaccatgg agacccttgg ggagcatgac ccctggtact
     3421 gtcccacctg taagaagcac cagcaggcaa caaagaagtt tgacttgtgg tctttgccca
     3481 agatcctggt ggttcacctc aagcgtttct cctataacag atactggcgg gataaacttg
     3541 acaccgtggt ggagttccca gtcagagctc tgaacatgtc cgagtttgtc tgtgaccggt
     3601 cagcaaggcc ttatgtttat gacctaattg ctgtgtccaa tcactatgga gccatggggg
     3661 ttggtcacta cactgcatat gcgaagaaca gactgaacgg gaaatggtat tactttgatg
     3721 atagcagtgt gtccctggcc tctgaggacc agatagtgac gaaagccgcc tacgtgttgt
     3781 tctatcagcg tcgggatgac gaatgctcca gcacatcttc gctcggcagt ttccccggtt
     3841 ctgatggagg ggtgaagctg agcagctcac accagggcat gggggatgag gaggcttaca
     3901 acatggacac caacacgcgt acgcggccgc tcgagcagaa actcatctca gaagaggatc
     3961 tggcagcaaa tgatatcctg gattacaagg atgacgacga taaggtttaa acggccggcc
     4021 gcggtcatag ctgtttcctg aacagatccc gggtggcatc cctgtgaccc ctccccagtg
     4081 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     4141 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     4201 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     4261 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     4321 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     4381 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     4441 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     4501 tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac
     4561 gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg
     4621 cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg
     4681 cggccagcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4741 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4801 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4861 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4921 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4981 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5041 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5101 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5161 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5221 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5281 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5341 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5401 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5461 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5521 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc
     5581 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg
     5641 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc
     5701 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg
     5761 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt
     5821 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg
     5881 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     5941 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     6001 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     6061 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     6121 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcgatccgc cacacccagc
     6181 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     6241 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
     6301 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     6361 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     6421 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     6481 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     6541 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     6601 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg
     6661 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     6721 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     6781 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     6841 tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     6901 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     6961 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     7021 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     7081 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     7141 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc
     7201 tcaggactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7261 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7321 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     7381 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     7441 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     7501 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     7561 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     7621 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     7681 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     7741 atttaacaaa aatttaacgc gaatttt
//
