LOCUS       V042220                 7651 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042220
VERSION     V042220
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             964..3141
                     /label="BRD3(NM_007371)"
                     /note="BRD3(NM_007371)"
                     /gene="BRD3"
     CDS             3148..3213
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3251..3475
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3521..3949
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3963..4292
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4359..5153
                     /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    5327..5448
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5497..5513)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5521..5537
                     /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(5545..5575)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5590..5611
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5899..6484)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6655..7515)
                     /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(7516..7620)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatgtcca ccgccacgac agtcgccccc gcggggatcc cggcgacccc gggccctgtg
     1021 aacccacccc ccccggaggt ctccaacccc agcaagcccg gccgcaagac caaccagctg
     1081 cagtacatgc agaatgtggt ggtgaagacg ctctggaaac accagttcgc ctggcccttc
     1141 taccagcccg tggacgcaat caaattgaac ctgccggatt atcataaaat aattaaaaac
     1201 ccaatggata tggggactat taagaagaga ctagaaaata attattattg gagtgcaagc
     1261 gaatgtatgc aggacttcaa caccatgttt acaaattgtt acatttataa caagcccaca
     1321 gatgacatag tgctaatggc ccaagcttta gagaaaattt ttctacaaaa agtggcccag
     1381 atgccccaag aggaagttga attattaccc cctgctccaa agggcaaagg tcggaagccg
     1441 gctgcgggag cccagagcgc aggtacacag caagtggcgg ccgtgtcctc tgtctcccca
     1501 gcgaccccct ttcagagcgt gccccccacc gtctcccaga cgcccgtcat cgctgccacc
     1561 cctgtaccaa ccatcactgc aaacgtcacg tcggtcccag tccccccagc tgccgcccca
     1621 cctcctcctg ccacacccat cgtccccgtg gtccctccta cgccgcctgt cgtcaagaaa
     1681 aagggcgtga agcggaaagc agacacaacc actcccacga cgtcggccat cactgccagc
     1741 cggagtgagt cgcccccgcc gttgtcagac cccaagcagg ccaaagtggt ggcccggcgg
     1801 gagagtggtg gccgccccat caagcctccc aagaaggacc tggaggacgg cgaggtgccc
     1861 cagcacgcag gcaagaaggg caagctgtcg gagcacctac gctactgcga cagcatcctc
     1921 agggagatgc tatccaagaa gcacgcggcc tacgcctggc ccttctacaa gccagtggat
     1981 gccgaggccc tggagctgca cgactaccac gacatcatca agcacccgat ggacctcagc
     2041 accgtgaaaa ggaagatgga tggccgagag tacccagacg cacagggctt tgctgctgat
     2101 gtccggctga tgttctcgaa ttgctacaaa tacaatcccc cagaccacga ggttgtggcc
     2161 atggcccgga agctccagga cgtgtttgag atgaggtttg ccaagatgcc agatgagccc
     2221 gtggaggcac cggcgctgcc tgcccccgcg gcccccatgg tgagcaaggg cgctgagagc
     2281 agccgtagca gtgaggagag ctcttcggac tcaggcagct cggactcgga ggaggagcgg
     2341 gccaccaggc tggcggagct gcaggagcag ctgaaggccg tgcacgagca gctggccgcc
     2401 ctgtctcagg ccccagtaaa caaaccaaag aagaagaagg agaagaagga gaaggagaag
     2461 aagaagaagg acaaggagaa ggagaaggag aagcacaaag tgaaggccga ggaagagaag
     2521 aaggccaagg tggctccgcc tgccaagcag gctcagcaga agaaggctcc tgccaagaag
     2581 gccaacagca cgaccacggc cggcagacag ctgaagaaag gcggcaagca ggcatctgcc
     2641 tcctacgact cagaggaaga ggaggagggc ctgcccatga gctacgatga aaagcgccag
     2701 cttagcctgg acatcaaccg gctgcccggg gagaagctgg gccgggtagt gcacatcatc
     2761 caatctcggg agccctcgct cagggactcc aaccccgacg agatagaaat tgactttgag
     2821 actctgaaac ccaccacttt gcgggaactg gagagatatg tcaagtcttg tttacagaaa
     2881 aagcaaagga aaccgttctc agcaagcggg aagaaacagg cagccaagtc gaaagaggag
     2941 ctagctcagg aaaagaagaa ggagctggaa aagcgtctgc aggatgtcag cgggcagctg
     3001 agcagcagca agaagcccgc ccggaaagag aagcccggct cagcaccctc agggggcccg
     3061 tccaggctca gcagcagcag ctcctccgag tctgggagca gcagctccag cgggtccagc
     3121 tctgacagca gtgactcaga actcgaggac tacaaagacc atgacggtga ttataaagat
     3181 catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta aacccgctga
     3241 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3301 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3361 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3421 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3481 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3541 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3601 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3661 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3721 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3781 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3841 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3901 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3961 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4021 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4081 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4141 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4201 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4261 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4321 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4381 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4441 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4501 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4561 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4621 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4681 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4741 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4801 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4861 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4921 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4981 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5041 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5101 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5161 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5221 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5281 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5341 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5401 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5461 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5521 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5581 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5641 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5701 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6061 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6121 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6181 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6241 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6301 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6361 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6421 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6481 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6541 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6601 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6661 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6721 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6781 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6841 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6901 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6961 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7021 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7081 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7141 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7201 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7261 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7321 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7381 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7441 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7501 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7561 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7621 cacatttccc cgaaaagtgc cacctgacgt c
//
