LOCUS       V019429                 9371 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019429
VERSION     V019429
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             671..4051
                     /label="TRIM33(NM_015906)"
                     /note="TRIM33(NM_015906)"
                     /gene="TRIM33"
     CDS             4058..4123
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4147..4735
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4801..4849
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5051..5479
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5493..5822
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5889..6683
                     /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    6859..6980
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7029..7045)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7053..7069
                     /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(7077..7107)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7122..7143
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7431..8019)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8190..9050)
                     /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(9051..9155)
                     /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 taccgccacc atggcggaaa acaaaggcgg cggcgaggct gagagcggcg gcgggggcag
      721 cggcagcgcg ccggtaactg ccggggccgc cgggcccgcc gcgcaggagg cggagccgcc
      781 tctcaccgcg gtgctggtgg aggaggagga ggaggaaggc ggcagggccg gcgctgaggg
      841 cggcgcggcc gggcccgacg acgggggggt ggccgcggcc tcctcgggct cggcccaggc
      901 tgcttcatct cctgcggcct cagtgggcac tggagttgcc gggggcgcag tatcgacgcc
      961 ggctccagct ccagcctcgg ctcccgctcc gggtccctcg gcagggccgc ctcctggacc
     1021 gccagcctcg ctcctggaca cctgcgccgt gtgtcagcag agcttgcaga gccggcgtga
     1081 ggcggagccc aagctgctgc cctgtcttca ctccttctgc ctgcgctgcc tgcccgagcc
     1141 ggagcgccag ctcagcgtgc ccatcccggg gggcagcaac ggcgacatcc agcaagttgg
     1201 tgtaatacgg tgcccagtat gccgccaaga atgcagacag atagaccttg tggataatta
     1261 ttttgtgaaa gacacatctg aagctcctag cagttctgat gaaaaatcag aacaggtatg
     1321 tactagttgt gaagacaatg caagtgcagt tggcttttgt gtagaatgtg gagagtggct
     1381 atgtaagaca tgtatcgaag cacatcaaag agtaaaattt actaaagatc acttgatcag
     1441 gaagaaagaa gatgtctcag agtctgttgg agcatctggt caacgccctg ttttctgccc
     1501 tgtacacaaa caagaacagt tgaaactttt ctgtgaaaca tgtgatagat tgacatgtag
     1561 agactgtcag ctattggaac acaaagaaca taggtatcag tttttggaag aagcttttca
     1621 aaatcagaag ggtgcaattg agaatctact ggcgaaactt cttgagaaga agaattatgt
     1681 tcattttgca gctactcagg tgcagaatag gataaaagaa gtaaatgaga ctaacaaacg
     1741 agtagaacag gaaattaaag tggccatttt cacccttatc aatgaaatta ataagaaagg
     1801 aaaatctctc ttacaacagc tagagaatgt tacaaaggaa agacagatga agttactaca
     1861 gcagcagaat gacatcacag gcctttcccg gcaggtgaag catgttatga acttcacaaa
     1921 ttgggcaatt gcaagtggca gcagcacagc actactatac agcaagcgac tgattacttt
     1981 ccagttgcgt catattttga aagcacggtg tgatcctgtc cctgctgcta atggagcaat
     2041 acgtttccat tgtgatccca ccttctgggc aaagaatgta gtcaatttag gtaatctagt
     2101 aatagagagt aaaccagctc ctggttatac tcctaatgtt gtagttgggc aagttcctcc
     2161 agggacaaac cacattagta aaacccctgg acagattaac ttagcacagc ttcgactcca
     2221 gcacatgcaa caacaagtat atgcacagaa acatcagcag ttgcaacaga tgaggatgca
     2281 gcaaccacca gcacctgtac caactacaac aacaacaaca caacagcatc ctagacaagc
     2341 agcccctcag atgttacaac aacagcctcc tcgattgatc agtgtgcaaa caatgcaaag
     2401 aggcaacatg aactgtggag cttttcaagc ccatcagatg agactggctc agaatgctgc
     2461 cagaatacca gggataccca ggcacagcgg ccctcaatat tccatgatgc agccacacct
     2521 ccaaagacaa cactcaaacc cagggcatgc tggacccttt cccgtagtat cggtacacaa
     2581 caccacaatc aacccaacga gccctactac agcaactatg gcaaatgcaa accgaggtcc
     2641 caccagccca tctgttacag caatagagct aatcccctca gttaccaatc cagaaaacct
     2701 tccatcgctg ccagatattc cacccataca gttggaagat gctggctcaa gtagtttaga
     2761 taatctacta agtagataca tctcaggcag tcacctaccc ccacagccta caagcaccat
     2821 gaatccttct ccaggtccct ctgccctttc tccgggatca tcaggtttat ccaattctca
     2881 cacacctgtg agacccccaa gtacttctag tactggcagt cgaggcagct gtgggtcatc
     2941 aggaagaact gctgagaaga caagtcttag tttcaaatct gatcaggtga aggtcaagca
     3001 agaacctggg actgaagatg aaatatgtag cttttcagga ggtgtaaaac aagaaaaaac
     3061 agaggatggc aggaggagtg cttgcatgtt gagcagtcct gagagtagct tgacaccacc
     3121 tctctcaacc aacctgcatc tagaaagtga attggatgca ttggcaagcc tggaaaacca
     3181 tgtgaaaatt gaacctgcag atatgaatga aagctgcaaa cagtcagggc tcagcagcct
     3241 tgttaatgga aagtccccaa ttcgaagcct catgcacagg tcggcaagga ttggaggaga
     3301 tggcaacaat aaagatgatg acccaaatga agactggtgt gctgtctgcc aaaacggagg
     3361 agatctcttg tgctgcgaaa aatgtccaaa ggtctttcat ctaacttgtc atgttccaac
     3421 actacttagc tttccaagtg gggactggat atgcacattt tgtagagata ttggaaagcc
     3481 agaagttgaa tatgattgtg ataatttgca acatagtaag aaggggaaaa ctgcgcaggg
     3541 gttaagcccc gtggaccaaa ggaaatgtga acgtcttctg ctttacctct attgccatga
     3601 attaagtatt gaattccagg agcctgttcc tgcttcgata ccaaactact ataaaattat
     3661 aaagaaacca atggatttat ccaccgtgaa aaagaagctt cagaaaaaac attcccaaca
     3721 ctaccaaatc ccggatgact ttgtggccga tgtccgtttg atcttcaaga actgtgaaag
     3781 gtttaatgaa atgatgaaag ttgttcaagt ttatgcagac acacaagaga ttaatttgaa
     3841 ggctgattca gaagtagctc aggcagggaa agcagttgca ttgtactttg aagataaact
     3901 cacagagatc tactcagaca ggaccttcgc acctttgcca gagtttgagc aggaagagga
     3961 tgatggtgag gtaactgagg actctgatga agactttata cagccccgca gaaaacgcct
     4021 aaagtcagat gagagaccag tacatataaa gctcgaggac tacaaagacc atgacggtga
     4081 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgatat
     4141 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4201 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4261 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4321 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4381 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4441 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4501 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     4561 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4621 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4681 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     4741 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4801 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     4861 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4921 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4981 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     5041 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5101 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     5161 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     5221 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     5281 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5341 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5401 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     5461 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     5521 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     5581 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5641 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5701 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5761 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5821 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     5881 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5941 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     6001 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     6061 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     6121 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     6181 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     6241 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     6301 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     6361 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     6421 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     6481 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     6541 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     6601 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     6661 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     6721 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6781 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6841 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6901 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6961 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     7021 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     7081 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     7141 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     7201 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     7261 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7321 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7381 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7441 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7501 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7561 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7621 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7681 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7741 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7801 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7861 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7921 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7981 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     8041 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     8101 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     8161 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     8221 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     8281 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     8341 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     8401 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     8461 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     8521 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     8581 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     8641 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     8701 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8761 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8821 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8881 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8941 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     9001 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     9061 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     9121 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     9181 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct
     9241 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     9301 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     9361 atctgcttag g
//
