LOCUS       V029906                 8460 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029906
VERSION     V029906
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             756..3140
                     /label="DHX15(NM_001358)"
                     /note="DHX15(NM_001358)"
                     /gene="DHX15"
     CDS             3147..3212
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3236..3824
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3890..3938
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4140..4568
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4582..4911
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4978..5772
                     /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    5948..6069
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6118..6134)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6142..6158
                     /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(6166..6196)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6211..6232
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6520..7108)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7279..8139)
                     /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(8140..8244)
                     /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 ctcgaattcg ccaccatgtc caagcggcac cggttggacc
      781 taggggagga ttacccctct ggcaagaagc gtgcggggac cgatgggaag gatcgagatc
      841 gagaccggga tcgtgaagat cggtctaaag atcgagaccg agaacgtgat agaggagata
      901 gagagcgaga gagggagaaa gaaaaggaga aggagttgcg agcttcaaca aatgctatgc
      961 ttatcagtgc tggattacca cctttgaaag cttcccattc agctcactca acccactcag
     1021 cacattcaac gcattcaaca cattctgctc attcaacgca tgccggacat gcaggtcaca
     1081 cgtcacttcc acagtgcatt aatccgttca ccaacttacc ccatactcct cgatactatg
     1141 atattctaaa gaaacgtctt cagctccctg tttgggaata caaggatagg tttacagata
     1201 ttctggttag acatcagtcc tttgtactgg ttggtgagac tgggtctggt aaaacaacac
     1261 agattccaca gtggtgtgtg gagtacatgc gatcattacc aggacccaag agaggagttg
     1321 cctgtaccca acccaggaga gtggctgcaa tgagtgtggc tcagagagtt gctgatgaga
     1381 tggatgtgat gttgggccag gaagttggtt actccattcg atttgaagac tgcagtagtg
     1441 caaaaaccat tcttaagtat atgactgatg ggatgttact tcgtgaagct atgaatgatc
     1501 ccctcctgga gcgttatggt gtaataattc ttgatgaggc tcatgagagg acactggcta
     1561 cagatattct aatgggtgtt ctgaaggaag ttgtaagaca gagatcagat ttaaaggtta
     1621 tagttatgag cgctactcta gatgcaggaa aattccagat ttactttgat aactgtcctc
     1681 tcctaactat tcctgggcgt acacatcctg ttgagatctt ctatactcca gaaccagaga
     1741 gagattatct tgaagcagca attcgaacag ttatccagat tcatatgtgt gaagaggaag
     1801 agggagatct tcttcttttc ttaactggtc aagaggaaat tgatgaagcc tgtaagagaa
     1861 taaagcgtga agttgatgat ttgggccctg aagttggtga cattaaaatc attccattgt
     1921 attctacact tccacctcag cagcagcaac gcatttttga gcctccacct cccaaaaaac
     1981 agaatggagc aattggaaga aaggtagttg tgtcaactaa catagcagag acgtctttga
     2041 caatagatgg tgtggtgttt gtgattgatc ctggatttgc gaaacagaag gtctacaatc
     2101 ctcgaatcag agttgagtcc cttttggtga cagctattag taaagcttca gctcagcaaa
     2161 gggctggtcg agctggacgt accagacctg gaaaatgctt cagactttac acagagaaag
     2221 cttataaaac agaaatgcag gataacacct atcctgagat tttgcgttct aatttaggat
     2281 cagttgtgtt acaattgaag aaacttggta ttgatgactt ggtacatttt gattttatgg
     2341 atccaccagc tcctgaaact ctgatgagag ccctggaact tttgaattac ctggctgctt
     2401 taaatgatga tggagatctg actgaattgg gatccatgat ggcagagttt cctctagatc
     2461 cacagctcgc aaaaatggtt attgcaagtt gtgactacaa ctgttctaat gaggtcctat
     2521 ctattactgc tatgttgtca gtcccacagt gttttgttcg ccccacggag gccaagaaag
     2581 ccgcagatga ggccaagatg agatttgccc acatagatgg agatcatctg acactgctga
     2641 acgtctacca tgcttttaaa caaaatcatg aatcggttca gtggtgttat gacaacttca
     2701 ttaactacag gtccctgatg tccgcagaca atgtacgcca gcagctatct cgaattatgg
     2761 acagatttaa tttgcctcgt cgaagtactg actttacaag cagggactat tatattaata
     2821 taagaaaagc tttggttact gggtatttta tgcaggtggc acatttagaa cgaacagggc
     2881 attacttaac tgtgaaagat aaccaggtgg ttcagttgca tccctctact gttcttgacc
     2941 acaaacctga atgggtgctt tataatgagt ttgttctaac aacaaagaat tacatccgga
     3001 catgtacaga tatcaagcca gaatggttgg tgaaaattgc ccctcaatat tatgacatga
     3061 gcaatttccc acagtgtgaa gcaaagagac agttggaccg catcattgcc aaacttcaat
     3121 ccaaggaata ttcacagtac ggatccgact acaaagacca tgacggtgat tataaagatc
     3181 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     3241 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3301 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3361 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3421 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3481 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3541 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3601 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3661 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3721 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3781 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     3841 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3901 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3961 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4021 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4081 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     4141 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4201 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4261 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     4321 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4381 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4441 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4501 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4561 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4621 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4681 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4741 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4801 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4861 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4921 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4981 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5041 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5101 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5161 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5221 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5281 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5341 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5401 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5461 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5521 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5581 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5641 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5701 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5761 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     5821 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5881 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5941 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6001 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6061 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     6121 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6181 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6241 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6301 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6361 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6421 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6481 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6541 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6601 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6661 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     6721 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6781 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6841 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6901 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6961 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7021 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7081 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7141 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7201 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7261 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7321 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7381 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7441 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7501 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7561 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7621 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7681 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7741 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7801 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7861 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7921 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7981 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8041 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8101 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8161 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8221 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     8281 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     8341 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     8401 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
