LOCUS       V045808                 8526 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045808
VERSION     V045808
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..3206
                     /label="Gtf3c4(NM_172977)"
                     /note="Gtf3c4(NM_172977)"
                     /gene="Gtf3c4"
     CDS             3213..3278
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3302..3890
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3956..4004
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4206..4634
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4648..4977
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5044..5838
                     /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    6014..6135
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6184..6200)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6208..6224
                     /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(6232..6262)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6277..6298
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6586..7174)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7345..8205)
                     /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(8206..8310)
                     /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 ccaccatgag cgaggcggac caggccctgg
      781 tggggcccaa ggccgacgag ccctcgccgc ccgcagagga gaaggacgaa gggggcggga
      841 aggaggcggc tgccgacgcg gcccccgggc ccagcgcctc gtttcgcctc atggtgactc
      901 ggcgggagcc tgccgtgaag ctgcaatacg cggtgagcgg gttagagccg ctctcctggt
      961 ctgaggacca ccgcgtgtct gtgtccacgg ctcgcagcgt cgccgtgctg gagcttatct
     1021 gcgatgtgca caacccaggc caggatctgg tgatccaccg cacctcggta cccgcacctc
     1081 tcaacagctg cctcctcaag gttggctcaa aaacagaagt tgctgagtgc aaggaaaagt
     1141 tcgcttcctc taaagacccc accatcagcc agactttcat gctagacagg atgttcaacc
     1201 ccgaggggaa ggccttgccc ccaatgcggg gtttcaaata cactagctgg tctccaatgg
     1261 gttgtgatgc aaatggcagg tgcctcttgg cagcactgac catggacaat cgcctgactg
     1321 tgcaggtgaa cctcaacaga ctccagtggg ttcagctggt tgatctgacg gagatttatg
     1381 gagaccgtct ttatgagact agttatagac tgtctaaaaa tgaggctcct gagggaaatc
     1441 tcggggattt tgctgagttt cagaggagac acagtatgca gaccccagtc agaatggaat
     1501 ggtcgagtat ctgtaccact cagcaggtca agcacaacaa cgagtgccgg gatgtgagca
     1561 gcgtgctgct ggctgtcctc tttgagaacg ggaatatagc tgtgtggcag ttccagctgc
     1621 cattcgtggg gaaggagtcc atctcttcat gcaacactat tgagtcagga atcagctccc
     1681 ccagtgtttt gttttggtgg gaatatgagc acaataatag gaaaatgagt ggccttattg
     1741 tgggcagtgc ttttggacct gtaaaaattc tccctgtcaa tctcaaagca gtgaaaggct
     1801 actttacttt aaggcagcct gttgtcttgt ggaaagaaat ggacaagtta cctgtgcaca
     1861 gcatcaaatg tgtgccactt taccacccct accagaagtg tagctgtagc ttagtggtag
     1921 ctgcaagagg ctcctatgtg ttttggtgtc ttcttctgat ctccaaggca ggtctgaatg
     1981 ttcacaattc ccacgtcaca ggcctccact cactgcccat cgtctccatt actgcagaca
     2041 agcagaatgg aacagtgtac acctgctcta gtgatgggaa ggtgaggcag ttgattccca
     2101 ttttcacaga tgttgcatta aagtttgaac accagctgat taagctctca gatgtctttg
     2161 gctcagtgag gactcacggc attgcagtga gtccctgtgg tgcttatctg gccatcatca
     2221 ccactgaggg catgatgaat ggcctccatc ctgtgaataa gaactaccag gtccagtttg
     2281 ttactctgaa gacctttgaa gaggcagctg ctcagctctt ggagtcttca gttcagaatc
     2341 tctttaagca ggtagacttg atcgacctgg tccgctggaa aattttaaaa gataaacata
     2401 ttcctcagtt tttacatgaa gcattggaaa aaaagattga aagcagcggg gttacctatt
     2461 tttggcgttt taagcttttc cttctaagga ttttgtatca gtcaatgcag aaatcccctt
     2521 ccgaggcctt atggaaaccc acccatgagg actcaaaaat cttactggtt gactcacctg
     2581 ggatgggcga cggtgaggat gagcagcaag aagaaggtac ttccaagcag gggactaagg
     2641 ctggcctgca ggagaagagc aaagaaggtg acacagagga gacccctgag gactcactca
     2701 ctgcaggggg agacaccgga ggccgtgaac ctgtagagga aaagctcctt gagatccaag
     2761 ggaagattga ggctgtggag atgcacctga ccagggagca catgaagcgg gtcctgggag
     2821 aagtgtacct gcacacctgg atcacagaga acaccagcat acccaccaga gggctctgta
     2881 acttcctgat gtctgatgag gattatgatg acaggacagc acaggtcttg attggacata
     2941 tctccaagaa gatgaacaaa cagaccttcc ctgagcgctg cagtctgtgt aaggagatct
     3001 taccattcac agatcgcaag caagctgttt gttccaatgg gcacatctgg ctccggtgct
     3061 ttctaaccta ccagtcctgc cagagcttga tatacagacg gtgtttgctc catgacagca
     3121 tcgccaggca tcctgttcca gaagatcctg actggattaa gaggttgctg cagagtcctt
     3181 gtcctttctg tgattctcct gtcttcggat ccgactacaa agaccatgac ggtgattata
     3241 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     3301 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3361 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3421 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3481 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3541 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3601 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3661 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     3721 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3781 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3841 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     3901 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3961 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     4021 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4081 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4141 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     4201 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4261 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4321 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     4381 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4441 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4501 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4561 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4621 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4681 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4741 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4801 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4861 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4921 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4981 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     5041 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5101 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5161 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5221 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5281 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5341 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5401 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5461 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5521 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5581 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5641 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5701 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5761 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5821 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     5881 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5941 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6001 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6061 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6121 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6181 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6241 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6301 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6361 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6421 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6481 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6541 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6601 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6661 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6721 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6781 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6841 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6901 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6961 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7021 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7081 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7141 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7201 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7261 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7321 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7381 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7441 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7501 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7561 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7621 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7681 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7741 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7801 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7861 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7921 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7981 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8041 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8101 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8161 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8221 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8281 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8341 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     8401 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     8461 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     8521 cttagg
//
