LOCUS       V033639                 7751 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033639
VERSION     V033639
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             890..934
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /transl_table=1
                     /translation="KETAAAKFERQHMDS"
                     /label="S-Tag"
     CDS             941..1054
                     /codon_start=1
                     /note="selected from a peptide library; binds streptavidin
                     with nanomolar affinity (Keefe et al., 2001)"
                     /product="streptavidin-binding peptide"
                     /transl_table=1
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
                     /label="SBP"
     CDS             1061..1087
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1094..3271
                     /label="BRD3(NM_007371)"
                     /note="BRD3(NM_007371)"
                     /gene="BRD3"
     polyA_signal    3318..3542
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3588..4016
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4030..4359
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4426..5220
                     /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    5394..5515
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5564..5580)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5588..5604
                     /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(5612..5642)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5657..5678
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5966..6551)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6722..7582)
                     /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(7583..7687)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttaccatga aagaaaccgc
      901 tgctgctaaa ttcgaacgcc agcacatgga cagcggtgga atggacgaga agaccaccgg
      961 ctggagaggc ggccacgtgg tggagggcct ggccggcgag ctggagcagc tgagagccag
     1021 actggagcac caccctcagg gccagagaga acctggtacc tacccatacg atgttccaga
     1081 ttacgctgga tccatgtcca ccgccacgac agtcgccccc gcggggatcc cggcgacccc
     1141 gggccctgtg aacccacccc ccccggaggt ctccaacccc agcaagcccg gccgcaagac
     1201 caaccagctg cagtacatgc agaatgtggt ggtgaagacg ctctggaaac accagttcgc
     1261 ctggcccttc taccagcccg tggacgcaat caaattgaac ctgccggatt atcataaaat
     1321 aattaaaaac ccaatggata tggggactat taagaagaga ctagaaaata attattattg
     1381 gagtgcaagc gaatgtatgc aggacttcaa caccatgttt acaaattgtt acatttataa
     1441 caagcccaca gatgacatag tgctaatggc ccaagcttta gagaaaattt ttctacaaaa
     1501 agtggcccag atgccccaag aggaagttga attattaccc cctgctccaa agggcaaagg
     1561 tcggaagccg gctgcgggag cccagagcgc aggtacacag caagtggcgg ccgtgtcctc
     1621 tgtctcccca gcgaccccct ttcagagcgt gccccccacc gtctcccaga cgcccgtcat
     1681 cgctgccacc cctgtaccaa ccatcactgc aaacgtcacg tcggtcccag tccccccagc
     1741 tgccgcccca cctcctcctg ccacacccat cgtccccgtg gtccctccta cgccgcctgt
     1801 cgtcaagaaa aagggcgtga agcggaaagc agacacaacc actcccacga cgtcggccat
     1861 cactgccagc cggagtgagt cgcccccgcc gttgtcagac cccaagcagg ccaaagtggt
     1921 ggcccggcgg gagagtggtg gccgccccat caagcctccc aagaaggacc tggaggacgg
     1981 cgaggtgccc cagcacgcag gcaagaaggg caagctgtcg gagcacctac gctactgcga
     2041 cagcatcctc agggagatgc tatccaagaa gcacgcggcc tacgcctggc ccttctacaa
     2101 gccagtggat gccgaggccc tggagctgca cgactaccac gacatcatca agcacccgat
     2161 ggacctcagc accgtgaaaa ggaagatgga tggccgagag tacccagacg cacagggctt
     2221 tgctgctgat gtccggctga tgttctcgaa ttgctacaaa tacaatcccc cagaccacga
     2281 ggttgtggcc atggcccgga agctccagga cgtgtttgag atgaggtttg ccaagatgcc
     2341 agatgagccc gtggaggcac cggcgctgcc tgcccccgcg gcccccatgg tgagcaaggg
     2401 cgctgagagc agccgtagca gtgaggagag ctcttcggac tcaggcagct cggactcgga
     2461 ggaggagcgg gccaccaggc tggcggagct gcaggagcag ctgaaggccg tgcacgagca
     2521 gctggccgcc ctgtctcagg ccccagtaaa caaaccaaag aagaagaagg agaagaagga
     2581 gaaggagaag aagaagaagg acaaggagaa ggagaaggag aagcacaaag tgaaggccga
     2641 ggaagagaag aaggccaagg tggctccgcc tgccaagcag gctcagcaga agaaggctcc
     2701 tgccaagaag gccaacagca cgaccacggc cggcagacag ctgaagaaag gcggcaagca
     2761 ggcatctgcc tcctacgact cagaggaaga ggaggagggc ctgcccatga gctacgatga
     2821 aaagcgccag cttagcctgg acatcaaccg gctgcccggg gagaagctgg gccgggtagt
     2881 gcacatcatc caatctcggg agccctcgct cagggactcc aaccccgacg agatagaaat
     2941 tgactttgag actctgaaac ccaccacttt gcgggaactg gagagatatg tcaagtcttg
     3001 tttacagaaa aagcaaagga aaccgttctc agcaagcggg aagaaacagg cagccaagtc
     3061 gaaagaggag ctagctcagg aaaagaagaa ggagctggaa aagcgtctgc aggatgtcag
     3121 cgggcagctg agcagcagca agaagcccgc ccggaaagag aagcccggct cagcaccctc
     3181 agggggcccg tccaggctca gcagcagcag ctcctccgag tctgggagca gcagctccag
     3241 cgggtccagc tctgacagca gtgactcaga atgactcgag tctagagggc ccgtttaaac
     3301 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3361 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3421 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3481 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3541 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3601 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3661 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3721 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3781 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3841 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3901 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3961 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4021 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4081 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4141 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4201 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4261 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4321 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4381 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4441 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4501 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4561 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4621 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4681 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4741 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4801 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4861 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4921 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4981 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5041 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5101 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5161 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5221 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5281 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5341 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5401 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5461 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5521 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5581 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5641 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5701 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5761 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5821 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5881 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5941 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6001 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6061 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6121 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6181 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6241 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6301 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6361 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6421 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6481 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6541 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6601 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6661 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6721 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6781 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6841 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6901 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6961 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7021 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7081 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7141 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7201 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7261 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7321 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7381 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7441 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7501 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7561 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7621 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7681 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     7741 tcccctatgg t
//
