LOCUS       V029809                 8318 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029809
VERSION     V029809
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..3808
                     /label="CHAF1A(NM_005483)"
                     /note="CHAF1A(NM_005483)"
                     /gene="CHAF1A"
     CDS             3815..3880
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3918..4142
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4188..4616
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4630..4959
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5026..5820
                     /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    5994..6115
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6164..6180)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6188..6204
                     /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(6212..6242)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6257..6278
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6566..7151)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7322..8182)
                     /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(8183..8287)
                     /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 atgctggagg agctggagtg
      961 cggggcgccc ggcgccaggg gagccgccac agccatggat tgcaaagata gaccagcttt
     1021 tccagttaag aagttaatac aagcccgtct gccgtttaag cgcctgaatc ttgtcccaaa
     1081 ggggaaagcc gatgacatgt cagacgatca gggtacttct gtgcaaagta aaagccccga
     1141 tttagaggcc tctttggaca ccttggaaaa caactgtcat gtgggttctg acatagactt
     1201 tagaccgaaa cttgtcaacg ggaagggtcc cttagataac tttttaagaa atagaatcga
     1261 aaccagtatt ggccagagca cagtcatcat tgatttgaca gaggactcga atgagcagcc
     1321 agacagtctt gtggaccaca ataaactaaa ttctgaagcc tctccctcca gggaggcaat
     1381 aaatggccag cgagaagaca ctggggatca gcaggggttg ttgaaggcca ttcagaacga
     1441 caagttggca tttcctggag agaccctttc agacattcct tgcaaaacag aggaggaggg
     1501 tgttggctgt ggaggtgcag ggaggagagg cgactcccag gaatgttcgc cacggagctg
     1561 cccggagctg acgagtggcc cgagaatgtg ccccagaaag gagcaggaca gttggagtga
     1621 agctgggggc atcctgttca aagggaaggt gcctatggtg gtcttgcagg acatcttggc
     1681 tgtgagacca ccgcaaatca agtcccttcc agccacaccc caaggcaaga acatgacccc
     1741 tgagagtgag gtgctggaat ctttccccga agaagactct gtactcagcc attcgtccct
     1801 gagctctccc tcttccacca gctcgcccga ggggccgcct gctcccccaa agcagcacag
     1861 cagtaccagt cccttcccca cctccacgcc cctccgcaga ataactaaga aattcgtcaa
     1921 aggctctaca gagaagaaca agctcagact gcaaagagat caggagcgtc tgggcaagca
     1981 gctcaagtta cgtgcagaaa gggaagaaaa ggagaagctg aaagaggagg ccaagcgggc
     2041 caaggaggag gccaagaaga agaaggagga agagaaggag cttaaggaaa aggagaggcg
     2101 ggagaagcgg gagaaggatg agaaggagaa ggcggagaag cagcggctca aggaggagcg
     2161 gcgcaaggag agacaggaag ccctggaggc taaacttgag gaaaaaagga aaaaggaaga
     2221 agagaaacgg ttaagagaag aagagaagcg cattaaagca gagaaggccg aaatcacgag
     2281 gttcttccag aaaccaaaga ctccacaggc ccccaagacc ctggccggct cctgtgggaa
     2341 gtttgccccc tttgaaatta aagagcacat ggtcctggcc cctcggcgtc ggaccgcttt
     2401 ccatccagac ctctgcagtc agctggacca gctcctccag cagcagagcg gcgagttctc
     2461 cttcttgaaa gacctcaaag gccggcagcc cctgaggtcc ggacccacgc acgtttccac
     2521 ccggaatgca gatattttta acagtgatgt cgtcatcgtg gagcgtggga agggcgacgg
     2581 tgttcccgag aggaggaagt ttggcaggat gaagctcctg cagttctgtg agaaccaccg
     2641 gcctgcctac tggggtacct ggaataagaa gacggcactc atccgcgcgc gagacccctg
     2701 ggcccaggac acgaagctcc tggactatga ggtggacagt gatgaggagt gggaagaaga
     2761 ggagcctggg gagtccctgt cccacagtga gggggatgat gatgacgaca tgggagagga
     2821 tgaagatgag gacgatggtt tctttgtgcc ccatgggtac ctgtctgagg acgaaggtgt
     2881 gacagaggag tgtgccgacc ctgagaacca taaggtccgc cagaaactga aggccaagga
     2941 gtgggacgag ttcctggcta aggggaagcg ctttcgcgtc ctgcaacctg tgaagatcgg
     3001 ctgcgtgtgg gcggctgaca gagactgcgc aggcgatgac ctgaaggtac tgcagcagtt
     3061 cgcagcctgc ttcctggaga ccctgccggc ccaggaggag cagacgccca aggcctccaa
     3121 gcgggagagg agagacgagc agatcctggc ccagctgctg ccgctcctgc acggcaatgt
     3181 gaacgggagc aaggtcatca tccgggagtt ccaggagcac tgccgccggg gactgctcag
     3241 caaccacacc ggcagcccgc ggagcccctc caccacctac ctgcacaccc ccacccccag
     3301 cgaggatgcc gccatcccct ctaagtcccg gctcaagcgg ctcatttccg agaactcagt
     3361 gtatgagaag cggcctgact tcaggatgtg ctggtacgtg cacccgcagg tgctacagag
     3421 cttccagcag gagcacctgc ccgtgccgtg ccagtggagc tatgtgacat cggtgccctc
     3481 ggcccccaaa gaggacagtg gcagcgtccc ctccacgggg cccagccagg gcactcccat
     3541 ctcgctgaag aggaagtcag cgggcagcat gtgcatcacc caattcatga agaagcgcag
     3601 gcacgacggc cagattggtg ctgaagacat ggacggcttc caggcagaca cggaggagga
     3661 ggaagaggag gagggcgact gtatgatcgt ggatgtcccg gatgctgcgg aggtccaagc
     3721 cccgtgtgga gccgcttccg gagctggggg tggtgtgggg gtggacaccg gcaaggccac
     3781 cctgaccgcg agcccactgg gtgcatccct cgaggactac aaagaccatg acggtgatta
     3841 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac
     3901 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3961 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     4021 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4081 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4141 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     4201 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4261 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4321 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4381 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4441 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4501 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4561 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4621 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4681 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4741 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4801 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4861 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4921 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4981 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5041 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5101 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5161 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     5221 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5281 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5341 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5401 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5461 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5521 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5581 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5641 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5701 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5761 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5821 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5881 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5941 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     6001 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6061 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6121 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6181 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6241 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6301 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6361 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6421 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6481 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6541 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6601 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6661 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6721 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6781 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6841 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6901 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6961 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     7021 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7081 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7141 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7201 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7261 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7321 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7381 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7441 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7501 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7561 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7621 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7681 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7741 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7801 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7861 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7921 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7981 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8041 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8101 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8161 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8221 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8281 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
