LOCUS       V018297                 8453 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018297
VERSION     V018297
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..3934
                     /label="GTF2I(NM_032999)"
                     /note="GTF2I(NM_032999)"
                     /gene="GTF2I"
     CDS             3950..4015
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4053..4277
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4323..4751
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4765..5094
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5161..5955
                     /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    6129..6250
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6299..6315)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6323..6339
                     /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(6347..6377)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6392..6413
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6701..7286)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7457..8317)
                     /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(8318..8422)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggcccaag ttgcaatgtc
      961 caccctcccc gttgaagatg aggagtcctc ggagagcagg atggtggtga cattcctcat
     1021 gtcagctctc gagtccatgt gtaaagaact ggccaagtcc aaagccgaag tggcctgcat
     1081 tgcagtgtat gaaacagacg tgtttgtcgt cggaactgaa agaggacgtg cttttgtcaa
     1141 taccagaaag gattttcaaa aagattttgt aaaatattgt gttgaagaag aagaaaaagc
     1201 tgcagagatg cataaaatga aatctacaac ccaggcaaat cggatgagtg tagatgctgt
     1261 agaaattgaa acactcagaa aaacagttga ggactatttc tgcttttgct atgggaaagc
     1321 tttaggcaaa tccacagtgg tacctgtacc atatgagaag atgctgcgag accagtcggc
     1381 tgtggtagtg caggggcttc cggaaggtgt tgcctttaaa caccccgaga actatgatct
     1441 tgcaaccctg aaatggattt tggagaacaa agcagggatt tcattcatca ttaagagacc
     1501 ttttttagag ccaaagaagc atgtaggtgg tcgtgtgatg gtaacagatg ctgacaggtc
     1561 aatactatct ccaggtggaa gttgtggccc catcaaagtg aaaactgaac ccacagaaga
     1621 ttctggtatt tccctggaaa tggcagctgt gacagtaaag gaagaatcag aagatcctga
     1681 ttattatcaa tataacattc aagcaggccc ttctgaaact gatgatgttg atgaaaaaca
     1741 gcccctatcg aagcctttgc aaggaagcca ccattcttca gagggcaatg aaggcacaga
     1801 aatggaagta ccagcagaag attctactca acatgtccct tcagaaacaa gtgaggaccc
     1861 tgaagttgag gtgactattg aagatgatga ttattctcca ccgtctaaga gaccaaaggc
     1921 caatgagcta ccgcagccac cagtcccgga acccgccaat gctgggaagc ggaaagtgag
     1981 ggagttcaac ttcgagaaat ggaatgctcg catcactgat ctacgtaaac aagttgaaga
     2041 attgtttgaa aggaaatatg ctcaagccat aaaagccaaa ggtccggtga cgatcccgta
     2101 ccctcttttc cagtctcatg ttgaagatct ttatgtagaa ggacttcctg aaggaattcc
     2161 ttttagaagg ccatctactt acggaattcc tcgcctggag aggatattac ttgcaaagga
     2221 aaggattcgt tttgtgatta agaaacatga gcttctgaat tcaacacgtg aagatttaca
     2281 gcttgataag ccagcttcag gagtaaagga agaatggtat gccagaatca ctaaattaag
     2341 aaagatggtg gatcagcttt tctgcaaaaa atttgcggaa gccttgggga gcactgaagc
     2401 caaggctgta ccgtaccaaa aatttgaggc acacccgaat gatctgtacg tggaaggact
     2461 gccagaaaac attcctttcc gaagtccctc atggtatgga atcccaaggc tggaaaaaat
     2521 cattcaagtg ggcaatcgaa ttaaatttgt tattaaaaga ccagaacttc tgactcacag
     2581 taccactgaa gttactcagc caagaacgaa tacaccagtc aaagaagatt ggaatgtcag
     2641 aattaccaag ctacggaagc aagtggaaga gatttttaat ttgaaatttg ctcaagctct
     2701 tggactcacc gaggcagtaa aagtaccata tcctgtgttt gaatcaaacc cggagttctt
     2761 gtatgtggaa ggcttgccag aggggattcc cttccgaagc cctacctggt ttggaattcc
     2821 acgacttgaa aggatcgtcc gcgggagtaa taaaatcaag ttcgttgtta aaaaacctga
     2881 actagttatt tcctacttgc ctcctgggat ggctagtaaa ataaacacta aagctttgca
     2941 gtcccccaaa agaccacgaa gtcctgggag taattcaaag gttcctgaaa ttgaggtcac
     3001 cgtggaaggc cctaataaca acaatcctca aacctcagct gttcgaaccc cgacccagac
     3061 taacggttct aacgttccct tcaagccacg agggagagag ttttcctttg aggcctggaa
     3121 tgccaaaatc acggacctaa aacagaaagt tgaaaatctc ttcaatgaga aatgtgggga
     3181 agctcttggc cttaaacaag ctgtgaaggt gccgttcgcg ttatttgagt ctttcccgga
     3241 agacttttat gtggaaggct tacctgaggg tgtgccattc cgaagaccat cgacttttgg
     3301 cattccgagg ctggagaaga tactcagaaa caaagccaaa attaagttca tcattaaaaa
     3361 gcccgaaatg tttgagacgg cgattaagga gagcacctcc tctaagagcc ctcccagaaa
     3421 aataaattca tcacccaatg ttaatactac tgcatcaggt gttgaagacc ttaacatcat
     3481 tcaggtgaca attccagatg atgataatga aagactctcg aaagttgaaa aagctagaca
     3541 gctaagagaa caagtgaatg acctctttag tcggaaattt ggtgaagcta ttggtatggg
     3601 ttttcctgtg aaagttccct acaggaaaat cacaattaac cctggctgtg tggtagttga
     3661 tggcatgccc ccgggggtgt ccttcaaagc ccccagctac ctggaaatca gctccatgag
     3721 aaggatctta gactctgccg agtttatcaa attcacggtc attagaccat ttccaggact
     3781 tgtgattaat aaccagctgg ttgatcagag tgagtcagaa ggccccgtga tacaagaatc
     3841 agctgaacca agccagttgg aagttccagc cacagaagaa ataaaagaga ctgatggaag
     3901 ctctcagatc aagcaagaac cagaccccac gtggatgcgg ccgctcgagg actacaaaga
     3961 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     4021 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4081 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4141 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4201 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4261 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4321 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4381 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4441 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4501 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4561 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4621 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4681 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4741 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4861 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4921 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4981 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5041 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5101 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5161 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5221 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5281 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5341 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5401 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5461 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5521 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5581 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5641 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5701 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5761 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5821 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5881 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5941 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6001 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6061 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6121 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6181 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6241 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6301 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6361 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6421 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6481 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6541 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6601 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6661 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6721 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6781 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6841 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6901 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6961 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7021 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7081 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7141 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7201 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     7261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7501 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7561 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7621 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7681 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7741 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7801 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7861 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7921 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7981 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8041 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8101 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
