LOCUS       V024386                10190 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024386
VERSION     V024386
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             779..4870
                     /label="Atad2(NM_027435)"
                     /note="Atad2(NM_027435)"
                     /gene="Atad2"
     CDS             4877..4942
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4966..5554
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5620..5668
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5870..6298
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6312..6641
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6708..7502
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    7678..7799
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7848..7864)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7872..7888
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(7896..7926)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7941..7962
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8250..8838)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9009..9869)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(9870..9974)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 ggtggtcctc cgcagcagcc tggagctgca cagtcgctcc accacctcct cggccaccga
      841 ctctctggac ctgtccagtg agttcctcag cctgcaggac agcggcggga ggcggcccag
      901 attgcgccga gccgggagga aatcgacggc gtccaaggcg gcgggcgatg gattcacagt
      961 taagggagct gaaacctacc acaacacgcg ctctttaagg tctctgagaa accaagcaca
     1021 aacttcttca gagtctagtt ttgacaagaa cgtggacaga acagagaaat actccaatgg
     1081 cagacatttc acaaggcaat ttgctcgaca gcaggctgat aaaaaagaag agtgcaaaga
     1141 agacaaagtt attccagtta ttcgctcatt gaggaataga aacattgctc caactgcaga
     1201 acatttgcac gaacaaagtg gagatgttga agttcgaaga agttgcagga ttagaagtcg
     1261 ttacagtacc atgaaccagt ctgtgctatt tgacaagctt ataacaaaca ctgctgaagc
     1321 tgtacttcaa aagatggatg acatgaaaaa gatgcgcaga cagcggatga agaagcttga
     1381 agacttggga gtattcaatg aaacagaaga gagtaatctc actatgtaca caagaggaaa
     1441 actaaaggct atccagcgag ctgatgaaga aacaactgat aaccaggacg gcagtgtgga
     1501 atcatctgaa gagggggaag agcaagagga tgatgatggc gaagatgaag atgatgaaga
     1561 tgaagaagag ggagaagaag ataatcaaaa aagatactat cttaggcaga gaaaaacaac
     1621 ggtttactac cagtctccac tggaaagtaa acctcgtcac cagagaaagc ccaacatgtt
     1681 ttatagtggc ccagcttctc ctgcaagacc aagattccgc ttgtcttcta cgggaccaag
     1741 aagtccctat tgtaaacgaa tgagcaggcg aaggcatgcc atccacagca gtgactctac
     1801 ttcctcctcc tcctctgaag acgactgttt tgagaggaga acaaaaagga accgaaatag
     1861 agctattaac aggtgcctcc cactaaattt tcggaaagat gaaataagag ggatttataa
     1921 agatcgaatg aaaattggag caagccttgc tgatgttgac ccaatgcaac tagatacttc
     1981 agtgcgcttt gacagtgttg gcggcctgtc cagtcacatt gcagctctaa aagagatggt
     2041 ggtgtttccc ttactttatc cagaagtctt cgaaaagttt aaaattcaac ccccaagagg
     2101 ttgtttgttt tatggaccac ctggaactgg aaaaaccctg gttgctagag cacttgccaa
     2161 tgagtgcagc cgaggggata agagagtggc gttcttcatg aggaaaggtg ctgactgcct
     2221 gagtaaatgg gtaggagaat ctgagagaca gcttcggttg ctatttgacc aggcctatca
     2281 gatgcgccca gcaattatat tctttgatga aattgatggt ttggctccag tacgatctag
     2341 caggcaagat cagattcaca gttctattgt ttcaaccctg ttagctctta tggatggttt
     2401 ggacagcaga ggagaaattg tggtcattgg agctaccaac cggctagatt ccatagaccc
     2461 tgctttacgg aggcccggtc ggtttgacag agaattcctt tttagtctac ctgataaaaa
     2521 tgctcgaaaa gagattctga agattcatac aagagattgg aatccaaagc cagtggacat
     2581 gtttctagaa gaactagcag aacactgtgt tggctactgt ggtgcggata ttaagtcaat
     2641 ctgtgctgaa gctgctttgt gcgccctgcg tcgacggtac ccgcagatct acaccaccag
     2701 cgagaagctt cagctggatc tctcttcaat taccatctca gctaaggatt ttgaggcagc
     2761 tctgcagaaa ataagaccag cttcccagag agctgtgaca tcacccggac aggcactgtc
     2821 tgccattgtg aaaccacttc tgcaaaatac tgttcatagg atcttagacg ccctacagaa
     2881 agtatttcca catgtggaag ttggaacaaa caaatcttta aattcagatg tttcttgccc
     2941 ttttctagaa agtgatttgg catacagtga tgatgataca ccatctgtgt atgaaaatgg
     3001 actttctcaa aaagaaaatt taaattttct tcatttaaat agaaatgcct gttaccagcc
     3061 tatgtctttt cgaccaagac tattgatagt gggagaacca ggatttggac agagttctca
     3121 cttggcacca gctgtcatcc atgctttgga aaaatttact gtatatacat tagacattcc
     3181 tgttcttttt ggcatcagta caacatctcc tgaagaggca tgttcccaaa tgattcgtga
     3241 agctaagaga acagcaccca gcatagtcta tgtcccacat atccatttgt ggtgggaaat
     3301 agttggaccg acactcaaag ccacatttac aacattatta cagactatac cttcatttgc
     3361 tccagtttta ttacttgcaa cttctgagaa accatactca gctttgccgg aagaggtaca
     3421 agaattgttt acccatgatt atggagaaat ttttaatgta cagttaccag ataaagaaga
     3481 acgaaccaaa ttttttgaag acttaatttt aaaacaagct tcaaagcctc ctgtgtcaca
     3541 aaagaaagca gttctgcagg ccttggaggt gctcccagtc gcaccaccgc ctgaaccaag
     3601 accactgaca gcagaggaag ttaaacggct tgaagaacaa gaagaagata cattcagaga
     3661 actcaggatt ttcttaagaa atgtcacaca tagacttgct attgataagc gattccgagt
     3721 ctttactaag cctgttgacc ctgatgaggt tcctgattat gtcactgtca taaagcaacc
     3781 aatggacctt tcatctgtaa tcagtaaaat tgatttacat aaatatctca ctgtgaaaga
     3841 ctatttgaag gatattgatc taatctgtag taatgcttta gaatacaatc cagatagaga
     3901 tcctggagat cgtcttatta ggcatagagc ttgtgctttg agagacactg cttatgcaat
     3961 aattaaagag gagcttgatg aagactttga gcagctctgt gaagaaattc aggaatctcg
     4021 gaagaagaga ggttgtagct cctccaaata cgcaccatct tattaccacg tcatgccaaa
     4081 gcaaaattct cctcctgttg gtgataaaaa gccagatcaa gagcagaatg agaagctaaa
     4141 ggtaccgtgc actcctgtgg cgtgcagcac acctgctcag ttgaaaagaa aattccataa
     4201 aaagtcaaag tggcatgtag gcaccaaaat aaagcgaagg aaaatttcac aagctaaaga
     4261 caacagcttg aatgctatga acagttcaag caggagcgac acagaagaca gccagcacac
     4321 acatgcagag cacacggagc ctggaaacac ggacgagtcc tcggtagaag aaagtgacaa
     4381 gcaaaataga cttgaaagca acatagactt gaaaaataac tcaagttcct ctaatattga
     4441 gaatgaactt gaagagccta aggaaactac ggagggcaca gaactgagaa aagacaggat
     4501 tgtttgcagg ggtgatgcct ctgcctccca ggtcacagac attcccgaag acagtgaatc
     4561 aaaagaaatg gattttctgc ggatgactct agctagaggt tcccaggtag agcagcagga
     4621 gctcatcagt atggagcagg ctctggccat cctctctcag ccgacaccct cacttgtgct
     4681 ggaccacaag cagttaacaa atattttgaa gacggttgtt aaaaaaagtc agaaatataa
     4741 tatattccaa ctggagaatt tgtatgcagt aatcagccag tgtatttatg agcaccgcag
     4801 ggactatgac aaaacagcgc ttgttcagaa aatggagcaa gcagtagaaa acttcaattg
     4861 ttccagatca ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga
     4921 ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca acctctggat
     4981 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     5041 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     5101 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     5161 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     5221 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     5281 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     5341 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     5401 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     5461 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     5521 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa
     5581 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     5641 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5701 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5761 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5821 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt
     5881 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5941 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     6001 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
     6061 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     6121 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     6181 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     6241 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     6301 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6361 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     6421 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     6481 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6541 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     6601 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     6661 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     6721 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     6781 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     6841 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     6901 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     6961 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     7021 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     7081 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     7141 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     7201 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     7261 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     7321 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     7381 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     7441 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     7501 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     7561 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     7621 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     7681 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     7741 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     7801 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     7861 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     7921 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     7981 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8041 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     8101 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     8161 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     8221 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     8281 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     8341 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     8401 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     8461 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     8521 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     8581 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     8641 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     8701 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     8761 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     8821 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     8881 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     8941 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     9001 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     9061 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     9121 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     9181 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     9241 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     9301 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     9361 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     9421 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     9481 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     9541 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     9601 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     9661 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     9721 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     9781 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     9841 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     9901 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     9961 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
    10021 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
    10081 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
    10141 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
