LOCUS       V039883                 8634 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039883
VERSION     V039883
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..3314
                     /label="TGS1(NM_024831)"
                     /note="TGS1(NM_024831)"
                     /gene="TGS1"
     CDS             3321..3386
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3410..3998
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4064..4112
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4314..4742
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4756..5085
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5152..5946
                     /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    6122..6243
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6292..6308)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6316..6332
                     /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(6340..6370)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6385..6406
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6694..7282)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7453..8313)
                     /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(8314..8418)
                     /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 ccaccatgtg ctgcgagaag tggagccgcg
      781 tggcggaaat gtttctcttc attgaggagc gggaggattg taagatactg tgcctttgct
      841 ccagggcatt tgtggaggat cgaaaattgt acaatttggg attaaaaggc tattacatca
      901 gagacagtgg caacaattca ggagaccagg cgacagaaga agaggaaggt ggttattcct
      961 gtggtactgc agaatcacat gacagcaaag gcataggcct ggatgaaagt gaacttgatt
     1021 ctgaggctga actcatgaga agtatgggat tgccacttca atttggtagg ataactgcac
     1081 ataaggattt tgaggtatct atgaatacta gaaataaagt taaaataaaa aagaaaaaac
     1141 atcaaaagaa atacttagat gaaattgtgc aagaatcttg gagaaaagaa tatgaagaag
     1201 acgacatttt ggcttcagat gatccatctt caattgaaca gtatgagaac accagaacat
     1261 atgaacttca aagcaaaaaa gatactgaga cagaaaatcc tccagttgaa aacacattat
     1321 ctccaaagct agaaattaca gagaaatggg aaaagtattg gaatgaatat ggaggaggac
     1381 tattgtggca aagttggcaa gaaaaacatc cgggtcaagc actatcttct gaaccttgga
     1441 actttcctga tacaaaggaa gaatgggagc aacattatag tcaactttat tggtattatt
     1501 tggaacaatt tcagtattgg gaagctcagg gttggacttt tgatgcctcg caaagctgtg
     1561 atacagatac ttacacatct aaaacagaag ctgatgacaa gaacgatgaa aaatgcatga
     1621 aagttgactt agtatctttt ccatcttcac ctattatggt tgataatgat agctctggta
     1681 caagtgataa ggatcatagt gaaatacttg atggaattag taacataaaa ctgaattcag
     1741 aggaagtaac acagagccaa ttagattcct gtacaagtca tgatggtcat caacagctaa
     1801 gtgaagttag tagcaaaaga gagtgccctg cttccggcca aagtgaacca cgtaatggag
     1861 gaaccaatga ggaaagcaac tcatcgggga atacaaacac agacccacca gctgaggatt
     1921 cacagaagtc ttcaggagca aacacaagca aagacagacc acatgccagt ggtactgatg
     1981 gagatgaaag tgaggaagac ccacctgagc ataagccaag caaactgaag aggagccatg
     2041 aactggacat tgatgaaaac ccagcttcag actttgatga cagtggttcc cttctaggat
     2101 tcaagtatgg ctcaggacaa aaatatggtg gaatcccaaa tttcagtcat cggcaggtca
     2161 ggtatttaga gaagaatgtg aagcttaagt ctaagtacct agacatgcgc agacaaataa
     2221 agatgaaaaa caaacacatc ttctttacca aagagtcaga aaaaccattt ttcaagaaaa
     2281 gcaaaattct gagtaaggta gaaaaattcc tcacatgggt taataaacca atggatgaag
     2341 aagcatcaca ggaatcatct tctcatgaca atgtgcacga cgcttccaca agtagtgatt
     2401 cagaggaaca agacatgtct gttaaaaaag gtgatgacct actggagact aataatccag
     2461 aacctgaaaa gtgtcagagc gtatcttcag ctggtgaact tgaaacagaa aactatgaaa
     2521 gagacagctt gctagcaact gttccagatg agcaggattg tgttactcaa gaagtgccag
     2581 actcccgcca ggcagaaact gaagctgaag tgaaaaagaa gaagaacaag aagaagaaca
     2641 aaaaggtgaa tggtctgcct cctgaaatag ctgctgttcc tgagctggca aaatactggg
     2701 cccagaggta caggctcttc tcccgttttg atgatgggat taagttggac agagagggct
     2761 ggttttcagt tacacccgag aagattgctg aacacattgc tggccgtgtt agtcagtcct
     2821 tcaagtgtga cgttgtagta gacgcattct gtggagttgg aggaaatacc attcagtttg
     2881 ccttaacagg aatgagagtg attgccattg atatcgatcc tgttaagatt gcccttgctc
     2941 gcaataatgc agaagtttat gggatagcag ataagataga gttcatctgt ggagatttct
     3001 tgctgctggc ttctttttta aaggctgatg ttgtgttcct cagcccacct tggggagggc
     3061 cagactatgc cactgcagag acctttgaca ttagaacaat gatgtctcct gatggctttg
     3121 aaattttcag actttctaag aagatcacta ataatattgt ttattttctt ccaagaaatg
     3181 ctgatattga ccaggtggca tccttagctg ggcctggagg gcaagtggaa atagaacaga
     3241 acttccttaa caacaaattg aagacaatca ctgcatattt tggtgaccta attcgaagac
     3301 cagcctctga aaccggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     3361 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     3421 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3481 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3541 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3601 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3661 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3721 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3781 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3841 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3901 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3961 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     4021 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4081 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4141 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4201 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4261 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     4321 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4381 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4441 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     4501 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4561 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4621 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4681 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4741 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4801 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4861 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4921 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4981 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     5041 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     5101 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     5161 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5221 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5281 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     5341 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5401 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5461 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5521 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5581 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5641 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5701 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5761 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5821 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5881 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5941 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     6001 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6061 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6121 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6181 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6241 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6301 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6361 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6421 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6481 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6541 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6601 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6661 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6721 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6781 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6841 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6901 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6961 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7021 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7081 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7141 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7201 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7261 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7321 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7381 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7441 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7501 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7561 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7621 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7681 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7741 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7801 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7861 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7921 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7981 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8041 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8101 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8161 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8221 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8281 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8341 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8401 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     8461 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     8521 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     8581 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
