LOCUS       V045363                10156 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045363
VERSION     V045363
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"
     CDS             706..4812
                     /label="DHX29(NM_019030)"
                     /note="DHX29(NM_019030)"
                     /gene="DHX29"
     CDS             4819..4908
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    4932..5520
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5586..5634
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5836..6264
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6278..6607
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6674..7468
                     /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    7644..7765
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7814..7830)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7838..7854
                     /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(7862..7892)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7907..7928
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8216..8804)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8975..9835)
                     /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(9836..9940)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggg cggcaagaac
      721 aagaaacaca aggctccagc ggccgcggtg gtccgggccg ccgtgtctgc ttccagagcc
      781 aaatctgccg aggctggaat tgccggggag gcccaaagca agaagccagt gtccaggccg
      841 gccaccgctg ccgctgccgc tgccggctcc agggagcccc gtgtcaagca aggtccaaaa
      901 atttatagtt ttaattctac aaatgattct agtggtcctg caaatctgga taaatctatt
      961 ttgaaagtgg taattaataa caaactagag caaagaatta ttggagtgat caatgagcat
     1021 aaaaagcaaa ataatgacaa aggaatgatt tctggaagac ttactgccaa aaaattgcag
     1081 gatttataca tggctttaca agcattttca tttaagacaa aggacattga agatgccatg
     1141 accaatacac tcttatatgg aggtgacctt cattctgcct tggattggct ctgtttaaac
     1201 ctttcagatg atgcacttcc tgaaggattc agtcaggaat ttgaagagca gcaacctaaa
     1261 agtaggccta aatttcagtc tcctcaaata caagccacta tttcacctcc attgcaacct
     1321 aaaacaaaaa catatgaaga ggaccctaag agtaagccaa aaaaggaaga aaaaaatatg
     1381 gaagtaaata tgaaagagtg gattttacga tatgctgaac aacaaaatga agaagaaaag
     1441 aatgagaatt ctaaaagttt agaagaggag gaaaaatttg accctaatga aaggtactta
     1501 catcttgcag caaaactgct ggatgcaaaa gaacaagcag ctacctttaa actagaaaaa
     1561 aacaagcaag gccaaaaaga ggctcaggaa aaaataagga aatttcaaag agaaatggaa
     1621 actttagaag accatccagt atttaaccca gccatgaaga tttcacatca acaaaatgaa
     1681 aggaaaaagc ctcctgtagc cacagaagga gaaagtgcat tgaattttaa tttatttgaa
     1741 aaatctgcag ctgctactga agaagagaaa gataaaaaga aagaacctca tgatgtaaga
     1801 aattttgact atactgctcg aagttggact ggaaaatctc ccaaacaatt tctgattgat
     1861 tgggtcagga agaatcttcc caagagtcca aatccttcct ttgaaaaagt tccagtaggt
     1921 agatactgga aatgtagggt tagggtaatc aagtctgaag atgatgtcct ggtagtatgc
     1981 cctacaatct taacagaaga tggcatgcaa gctcagcacc tgggagctac tttagccctt
     2041 taccgtttag ttaaagggca gtcagttcat cagttacttc ctcccactta ccgagatgtt
     2101 tggctggagt ggagtgatgc agaaaagaaa agggaagaat taaataaaat ggaaaccaat
     2161 aaaccacgtg atctttttat tgccaaactt ctgaataaac tgaaacagca gcaacagcag
     2221 caacaacagc attctgaaaa taagagagaa aactctgaag atcccgagga atcttgggaa
     2281 aatttagttt cggatgagga tttttctgca ctgtccttgg aatcagcaaa tgtggaagat
     2341 ttggaacctg ttagaaacct ctttagaaag ttgcaaagca cacctaagta tcagaaactt
     2401 ctaaaggaaa gacaacagct acctgtattt aaacatcggg actcaattgt tgaaactctt
     2461 aaaaggcatc gggtagtggt tgtggcaggt gaaacaggga gtggtaaaag tactcaggta
     2521 ccacattttc tattggaaga tttgcttcta aatgagtggg aagcaagtaa atgtaacatt
     2581 gtctgtaccc aaccccgaag aatctcagca gttagtttag ccaacagagt atgtgatgaa
     2641 ttgggctgtg aaaatggacc tggaggaagg aattccttgt gtggatatca gatccggatg
     2701 gaatctcgag cttgtgaatc taccaggtta ctctattgta caacaggggt tttgctaagg
     2761 aaacttcaag aagatggtct tctaagtaat gtgtctcatg ttattgtaga tgaggttcat
     2821 gaaagaagtg tccagtcaga cttcctacta attatcttga aggaaatttt acagaaacgt
     2881 tctgatctac acttgattct aatgagtgcc actgtggaca gcgaaaaatt ttctacatat
     2941 ttcacacact gccccattct cagaatttca ggaagaagtt atcctgttga ggtttttcat
     3001 cttgaagata taatagaaga aacaggcttt gtactggaaa aagactcaga atattgtcag
     3061 aaatttctgg aagaggaaga agaagtaacc attaatgtta caagcaaagc agggggaata
     3121 aaaaaatatc aggaatacat cccagttcag actggagcac atgctgattt aaatccattt
     3181 taccaaaagt acagcagccg cactcagcat gctattctat acatgaatcc tcataaaatc
     3241 aacctggatc tcattttgga acttcttgca tacttagata aaagtcccca attcagaaat
     3301 attgaaggag cagtattgat ctttttacca ggacttgctc atattcagca gttgtatgat
     3361 cttctatcaa atgatagaag attttattct gaacgatata aagtgatagc tctgcattct
     3421 attctttcaa cccaagatca agctgcagca ttcacacttc cccctccagg agtcaggaag
     3481 attgttttag caaccaatat tgcagagacg ggtatcacta ttcctgatgt tgtatttgta
     3541 attgatactg gaagaacaaa agaaaataag taccatgaaa gcagtcagat gagttctttg
     3601 gtggagacgt ttgtcagtaa agccagtgct ttgcagcgcc agggaagagc tgggcgggtc
     3661 agagatggct tctgtttccg aatgtacaca agagaaagat ttgaaggctt tatggattat
     3721 tctgttcctg aaatcttacg tgtacctttg gaggaattat gccttcatat tatgaaatgt
     3781 aatcttggtt ctcctgaaga tttcctctcc aaagccttag atcctcctca gctccaagtg
     3841 atcagcaatg caatgaattt gctccgaaaa attggagctt gtgaattaaa tgagcctaaa
     3901 ctgactccgt tgggccaaca ccttgcagct ttacctgtga atgtcaagat tggcaagatg
     3961 cttatttttg gtgccatatt tggctgcctt gacccagtgg caacactagc tgcagttatg
     4021 acagagaagt ctccttttac cacaccaatt ggtcgaaaag atgaagcaga tcttgcaaaa
     4081 tcagctttgg ccatggcgga ttcagaccac ctgacgatct acaatgcata tctaggatgg
     4141 aagaaagcac gacaagaagg aggttatcgt tctgaaatca catactgccg gaggaacttt
     4201 cttaatagaa catcactgtt aaccctagag gatgtaaagc aggagttaat aaagttggtt
     4261 aaggcagcag gattttcatc ttccacaact tctaccagct gggaaggaaa cagagcctca
     4321 cagaccctct cattccaaga aattgccctt cttaaagctg tactggtggc tggactgtat
     4381 gacaatgtgg ggaagataat ctatacaaag tcagtggatg ttacagaaaa attggcttgc
     4441 attgtggaga cggcccaagg caaagcacaa gtacacccat cctcagtaaa tcgagatttg
     4501 caaactcatg gatggctctt ataccaggag aagataaggt atgccagagt gtatttgaga
     4561 gaaactaccc taataacccc ttttccagtt ttactttttg gtggtgatat agaagttcag
     4621 caccgagaac gtcttctttc tattgatggc tggatctatt ttcaggcccc tgtaaagata
     4681 gctgtcattt tcaagcagct gagagttctc attgattcag ttttaagaaa aaagcttgaa
     4741 aatccaaaga tgtcccttga aaatgacaag attctgcaga tcattacgga attgataaaa
     4801 acagagaata acctcgagga acaaaaacta atatcagagg aagatttgga acagaaacta
     4861 atcagtgaag aagatttaga gcaaaagtta atttccgagg aggacttata gtgagggccc
     4921 gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     4981 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     5041 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     5101 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     5161 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     5221 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     5281 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     5341 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     5401 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     5461 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     5521 ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     5581 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac
     5641 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5701 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5761 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct
     5821 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5881 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5941 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt
     6001 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     6061 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     6121 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     6181 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga
     6241 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     6301 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     6361 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     6421 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6481 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6541 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     6601 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     6661 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     6721 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     6781 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     6841 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     6901 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     6961 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     7021 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     7081 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     7141 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     7201 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     7261 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     7321 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     7381 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     7441 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
     7501 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     7561 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     7621 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     7681 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     7741 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     7801 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     7861 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     7921 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7981 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     8041 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     8101 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     8161 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     8221 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     8281 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     8341 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     8401 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     8461 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     8521 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     8581 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     8641 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     8701 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     8761 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8821 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8881 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8941 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     9001 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     9061 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     9121 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     9181 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     9241 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     9301 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     9361 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     9421 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     9481 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     9541 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     9601 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     9661 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     9721 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     9781 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     9841 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9901 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9961 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat
    10021 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
    10081 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
    10141 gaagaatctg cttagg
//
