LOCUS       V027005                 7727 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027005
VERSION     V027005
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..3217
                     /label="Cul4a(NM_146207)"
                     /note="Cul4a(NM_146207)"
                     /gene="Cul4a"
     CDS             3224..3289
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3327..3551
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3597..4025
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4039..4368
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4435..5229
                     /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    5403..5524
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5573..5589)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5597..5613
                     /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(5621..5651)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5666..5687
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5975..6560)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6731..7591)
                     /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(7592..7696)
                     /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 atggcggacg agggccctcg
      961 gaagggcagc gtctcggctc tcatgggccg caccaacggc ctcaccaagc ccgcggcgtt
     1021 ggccggtggc cccgccaagc cggggggcac gggaggctcc aggaagctgg tcatcaagaa
     1081 cttccgagac aggccgcggc tgcctgacaa ctacactcag gacacgtggc ggaagcttca
     1141 cgaagcggtc aaggccatcc agagcagcac atccatcagg tacaacctgg aggagctgta
     1201 ccaggctgtt gaaaatcttt gttctcacaa agtctcccca acgctctaca agcagctgcg
     1261 ccaggtctgt gaagaccatg tccaggccca gatcctccca ttcagagaag actcactaga
     1321 cagcgtttta tttctaaaga agattaacac atgctggcaa gatcactgca gacaaatgat
     1381 catgatcaga agcatcttcc tgtttctgga tcgaacctac gtcctccaga attccatgct
     1441 gccttctatc tgggacatgg gcctagagct gtttaggaac cacatcatca gcgacaggat
     1501 ggtgcagagt aagaccattg atgggatcct gctgctgatt gggcgtgaac gaagtggaga
     1561 ggctgtggac aggagtctgc ttcggagttt gctaagcatg ctctcagatc tccaggtgta
     1621 caaagactca tttgagctga agttcttgga agagactaac tgtctatacg ctgctgaagg
     1681 ccagaggctg atgcaggaca gggaggttcc agaatacctt aaccatgtga gcaaacgtct
     1741 agaagaggaa gcagaccgag tgatcaccta cttagaccac agcacacaaa aaccgttgat
     1801 tgcctgtgtg gagaaacagc tgttaggaga acatttgaca gcaattctac agaaaggcct
     1861 agagcacctg ctggatgaga acagggtgcc cgacctcacc cagatgtacc agctcttcag
     1921 ccgggtgaaa ggagggcagc atgcgctgct gcagcactgg agcgagtaca tcaagacctt
     1981 tggaacaacc attgtcatca atcctgagaa agacaaagac atggtccaag acctgctgga
     2041 ctttaaggac aaagtggacc acgtggtgga ggtgtgcttc cagaggaacg agcgcttcat
     2101 caacctgatg aaggagtcct tcgagacgtt catcaacaag agaccaaaca agcctgcaga
     2161 gctcattgcc aagcacgtgg actcaaagtt acgggccggc aacaaagaag ccacagatga
     2221 ggaactggag aggatcctgg acaagatcat gattctcttc cgtttcatcc atggtaaaga
     2281 tgtctttgaa gcgttttata agaaagactt agcaaaaagg ctgctggtgg ggaagagcgc
     2341 gtcggtggac gcggagaagt ccatgctgtc gaagctcaag cacgagtgcg gagctgcgtt
     2401 taccagcaaa ctggagggca tgttcaagga catggagctc tccaaagaca tcatggttca
     2461 cttcaagcag catatgcaga accagagtgc gccaggcccc attgacctca cggtgaacat
     2521 actcaccatg ggctactggc ccacgtacac acccatggaa gtgcacctcc ctccagagat
     2581 ggtcagactt caggaagtat tcaagacgtt ctacctgggc aagcacagtg ggcggaagct
     2641 gcaatggcag acaacgctgg gacacgccgt gctgaaggcg gactttaagg aggggaagaa
     2701 ggagttccag gtatccctct tccagacgct ggtgctgctc atgttcaacg aaggggatgg
     2761 atttagcttt gaagaaataa aaatggccac tggcatagag gacagtgagt tgcgaagaac
     2821 actacagtct ctggcctgtg gtaaagcacg ggtcctaatt aaaagtccca aaggaaagga
     2881 ggtggaagat ggagataagt tcatcttcaa tgcagacttc aaacacaagt tgttcagaat
     2941 aaagatcaat caaatccaga tgaaggagac cgttgaagaa caggttagca ctacagagag
     3001 agtattccag gatagacagt accagatcga cgctgctata gtcagaataa tgaaaatgag
     3061 gaagacactt ggtcataatc ttctagtttc tgaattatac aatcagctga aatttccagt
     3121 gaagcctgga gatttgaaaa aaaggattga gtcacttata gacagagact atatggaacg
     3181 agacaaagac agtccaaatc agtaccacta cgtggcactc gaggactaca aagaccatga
     3241 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     3301 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3361 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3421 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3481 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3541 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3601 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3661 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3721 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3781 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3841 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3901 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3961 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4021 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4081 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4141 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4201 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4261 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4321 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4381 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4441 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4501 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4561 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4621 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4681 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4741 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4801 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4861 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4921 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4981 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5041 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5101 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5161 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5221 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5281 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5341 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5401 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5461 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5521 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5581 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5641 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5701 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5761 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5821 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5881 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5941 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6001 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6061 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6121 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6181 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6241 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6301 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6361 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6421 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6481 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6541 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6601 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6661 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6721 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6781 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6841 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6901 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6961 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7021 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7081 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7141 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7201 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7261 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7321 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7381 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7441 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7501 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7561 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7621 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7681 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
