LOCUS       V031213                 8133 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031213
VERSION     V031213
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..4280
                     /label="Tshz1(NM_001081300)"
                     /note="Tshz1(NM_001081300)"
                     /gene="Tshz1"
     CDS             4299..4328
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             4347..4370
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(4390..4406)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    4417..5039
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(5188..5776)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    6105..6152
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(6384..7178)
                     /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(7213..7570)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(7572..7676)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7703..8131
                     /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 gccgaggagg aagcagcagg ccccccggcg ctcggcagct tacgttcccg
     1081 aggaagaact gaaggcagcc gaaatagatg aggagcacgc ggaggatggt gggctgtctc
     1141 tggacattca ggagagcgaa tttgcgtgca atgaagagac agagatcaaa gaagcccaga
     1201 gctatcagaa ctcccctgtc agcactgcca ccaatcaaga cgctggctac ggatcgccct
     1261 tcagtgaggg cagtgaccag ctggcccatt tcaaaagctc ctcctccaga gaagagaagg
     1321 aggagtcgca gtgtccagac agtgtctcct acccccagga cagcttggca cagatcaaag
     1381 ccgtgtatgc gaacttattc tcagagtcct gctggtctag cctagcattg gatttaaaaa
     1441 agtccaattc tgccaccagc aacaacgatg ccagccagaa ggagagttcc acacctaccc
     1501 ccacgcctcc tactagtacc gccagcaccg cctgtaccac ggccaccacg gccatcacca
     1561 gctgcagcac cagcaccagc catagcagca ccaccagtaa cagcagcagc tcggggtatg
     1621 actggcacca ggccgccctg gccaagacgc tgcagcagac gtcgtcctac ggactgctgc
     1681 ctgagcccag cctgttcagc accgtgcagc tctaccgaca gaataacaag ctgtatggct
     1741 ccgtgttcac gggggccagc aagttcaggt gcaaagactg cagcgcggcc tacgacacgc
     1801 tggtggaact gacggtgcac atgaacgaga caggccacta tcgagatgac aacagagaca
     1861 aggactccga gaagaccaag cggtggtcca agcccaggaa gcgttccctg atggagatgg
     1921 aggggaagga ggacgcccag aaggtgctga agtgtatgta ctgcggacac tccttcgagt
     1981 ccttgcaaga cctcagtgtc cacatgatca aaacaaagca ttaccagaaa gtgcctctga
     2041 aggagcccgt gccagccatc accaaactgg tcccttccac caagaagcga gcactccagg
     2101 acctggcatc cccttgttcc ccagagccca cgggcgtggc cactgaggtt gcactgagtg
     2161 agtcagccaa ggaccagaaa acggccaacc cgtacgtgac tccgaacaac cgttacggct
     2221 accagaatgg tgctagttac acgtggcagt tcgaggcccg caaagcccag atactcaagt
     2281 gcatggaatg tggcagctcc catgatacct tgcagcagct cacagcccac atgatggtca
     2341 ctggccattt cttaaaggtg acaacctctg cctccaagaa aggaaagcag ctggtgttgg
     2401 accccgtggt ggaggagaag atccagtcca tcccgttgcc cccgaccact cacacccggc
     2461 tgcctgcctc tagcatcaag aagcaaccgg actctcctgc gggctctgta gcctcagaag
     2521 agaagaagga acctgagaag gagaaggaaa aggagaaagc cccaccggca gctggggatg
     2581 cagagagaaa aatcaaagag gagactgaag atgccactga gaagtttgag cccactgccc
     2641 tttatcagta cctgcgtgag gaagacctag atgacagccc caagggagga gtagatatcc
     2701 taaagtcact cgagaacact gtgtccacgg ccatcagtaa agcccagaat ggggcaccct
     2761 catggggtgg gtaccccagc atccatgcag cctaccagct gccaggcaca gtgaagccac
     2821 tgcagtctgc tgtgcagagc gtgcaaatac agccgtctta cgccagcagc gtgaagtcac
     2881 tgtcctccac agagcacaat gccctcctac actccccagg gagcctcacc ccgccacccc
     2941 ataagagcaa tgtgtctgcc atggaggagt tagtagagaa ggtcacgggc aaggtcagca
     3001 tcaagaagga ggagaggcct actgagaagg agaagagctc ccctgtcaaa gccatctctc
     3061 ctgtggcaaa ggagaataaa gatcttccta agacagagga gaccgggagc aagccacaga
     3121 agaagggctc ggattcagaa actgggaagg ccaaaaagga gagtacattg gacgctcaca
     3181 ccccaaatgg tacagaacct cttaaagcca aggtcaccaa tggctgcggc catctgggta
     3241 tcatcacgga ccactcgccg gagccttctt tcatcaaccc tctaagcgct ctgcagtcca
     3301 tcatgaacac ccacctagga aaggtgtcca agcccgtgag cccctctctg gatccactgg
     3361 cgatgctcta caagatcagc aacagcatgc tggacaagcc cgtctacccc acgacccctg
     3421 ccaagcaggc tgatgcaatc gaccgctact actatgagaa cagcgaccag cccattgatc
     3481 tgaccaagtc taagaacaag ccactggtct cgggtgtggc agatgctgtg tcctcgcctc
     3541 tgcgggagag cgccctcatg gatatctcgg acatggtgaa aaatctcaca ggtcgtctga
     3601 ctcccaagtc ctccacaccc tccacggtgt cagagaagtc agatgcagat gggagcagct
     3661 ttgaggaggc cctggacgag ctgtcacctg ttcacaagag aaaggggcgc cagtccaatt
     3721 ggaacccaca acacctgctc atcctgcagg ctcagtttgc ctccagcctt cgagagacag
     3781 cagaagggaa atacatcatg tcagacctgg gccctcagga gcgggtgcac atctccaagt
     3841 tcacgggcct atccatgacc accatcagtc actggctggc caacgtgaag taccagctgc
     3901 ggaggacagg gggaaccaag ttccttaaga acctcgacac agggcatcct gttttctttt
     3961 gtaatgattg tgcctctcag ttcagaactg cctccacgta cgtcagtcac ctggagacgc
     4021 acctaggctt cagcttgaag gacctctcca agctgccact cagtcagata caagagcagc
     4081 agagtgtcgt ctccaaagcc ctcaccaaca agactctggg cccgctgggc tcctccgagg
     4141 aagacctggg ctccacattc caatgcaagc tctgcaaccg gacttttgcg agcaagcacg
     4201 cagtaaaact gcaccttagt aagacgcacg gcaagtcgcc agaggaccac ctgatctacg
     4261 tgacagagct ggagaagcaa acgcgtacgc ggccgctcga gcagaaactc atctcagaag
     4321 aggatctggc agcaaatgat atcctggatt acaaggatga cgacgataag gtttaaacgg
     4381 ccggccgcgg tcatagctgt ttcctgaaca gatcccgggt ggcatccctg tgacccctcc
     4441 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     4501 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     4561 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     4621 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     4681 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     4741 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     4801 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     4861 caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact ataccagcag
     4921 gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg acatttgagt
     4981 tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc tgttgaattg
     5041 ggtacgcggc cagcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5101 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5161 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5221 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5281 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5341 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5401 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5461 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5521 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5581 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5641 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5701 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5761 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5821 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5881 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aacctgaggc
     5941 tatggcaggg cctgccgccc cgacgttggc tgcgagccct gggccttcac ccgaacttgg
     6001 ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg
     6061 ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa acgacccaac
     6121 accgtgcgtt ttattctgtc tttttattgc cgtcatagcg cgggttcctt ccggtattgt
     6181 ctccttccgt gtttcagtta gcctccccct agggtgggcg aagaactcca gcatgagatc
     6241 cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc
     6301 atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt
     6361 catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg
     6421 cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg
     6481 ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg atccgccaca
     6541 cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc
     6601 aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgct cgccttgagc
     6661 ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg
     6721 acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg
     6781 aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat
     6841 actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat
     6901 agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc
     6961 gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac
     7021 aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca
     7081 tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg
     7141 gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc
     7201 tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     7261 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     7321 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     7381 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     7441 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     7501 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
     7561 tccgcctcag gactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7621 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7681 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     7741 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     7801 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     7861 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     7921 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     7981 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     8041 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     8101 gagctgattt aacaaaaatt taacgcgaat ttt
//
