LOCUS       V027654                10596 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027654
VERSION     V027654
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..5276
                     /label="DCAF1(NM_014703)"
                     /note="DCAF1(NM_014703)"
                     /gene="DCAF1"
     CDS             5283..5348
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    5372..5960
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    6026..6074
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6276..6704
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6718..7047
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7114..7908
                     /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    8084..8205
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8254..8270)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8278..8294
                     /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(8302..8332)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8347..8368
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8656..9244)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9415..10275)
                     /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(10276..10380)
                     /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 ccaccatgac tacagtagtg gtacatgtgg
      781 actccaaagc tgagctcact accctgctgg agcagtggga aaaggaacat ggcagtgggc
      841 aggacatggt acctatcctt accaggatgt ctcaattgat tgaaaaagaa actgaagagt
      901 atcgtaaagg ggatccagac ccatttgatg atcgacatcc tggtcgagct gatccagagt
      961 gtatgctggg ccacttgctg agaatactct tcaagaatga tgatttcatg aatgcactgg
     1021 tgaatgcata tgtgatgaca agccgagagc cccctttaaa cactgcagct tgcagactcc
     1081 tattagacat catgccaggg ctggaaactg ctgtcgtctt tcaagaaaag gagggaattg
     1141 tcgagaatct tttcaaatgg gcccgagagg ccgatcaacc attgaggaca tattctactg
     1201 gactgttagg aggtgctatg gaaaatcaag acattgctgc caactataga gatgaaaatt
     1261 cacagctggt ggcaatagtg cttcgaagac tgagggagct acagctacag gaagtggctt
     1321 tgcggcagga aaacaagcgt cccagtccac ggaagctctc ttctgaaccc cttttgcctc
     1381 tggatgagga ggctgtggat atggactatg gtgacatggc tgtagatgta gtggatggag
     1441 accaagagga agcttctgga gacatggaga tctcctttca tcttgattca ggccacaaga
     1501 ctagtagcag agtgaactca acaaccaaac ctgaggatgg aggattaaag aaaaacaagt
     1561 cagcaaaaca gggtgacaga gagaacttta ggaaagccaa gcaaaagttg ggtttctcat
     1621 cttctgatcc agatcgcatg tttgttgagc tgtctaatag cagttggtca gaaatgtctc
     1681 cctgggtgat tggcaccaat tatacccttt atcctatgac tcctgctatc gagcagcgac
     1741 tcattctcca atatttgacc cctctaggag aatatcagga gctacttccc atattcatgc
     1801 aacttggatc acgggagctg atgatgttct atattgacct gaagcaaact aatgatgtcc
     1861 tgcttacatt tgaggcacta aagcacctag catctctcct gctccataac aaatttgcca
     1921 cagaatttgt tgcgcatggt ggagtacaga aattactgga aatacctcgt ccttctatgg
     1981 ctgcaaccgg tgtgtctatg tgtttgtatt acctatccta caatcaggat gccatggaaa
     2041 gagtttgcat gcatccccac aatgttctgt ctgatgtggt gaactatacc ctgtggttaa
     2101 tggagtgttc tcatgcttca ggatgctgcc atgctaccat gtttttttca atttgcttct
     2161 catttcgggc cgtcttggag ctctttgacc gctatgatgg tcttcgtcgt ctggtgaact
     2221 tgatcagtac tttggagatt ctaaatttgg aagatcaggg tgcacttctg agtgatgatg
     2281 aaatatttgc tagccgccaa actgggaaac atacctgcat ggccttgcgc aaatactttg
     2341 aggctcacct ggccattaaa ttggaacaag tgaagcagtc acttcagagg actgagggtg
     2401 gcattcttgt ccacccacaa cccccgtaca aggcatgctc atatactcat gaacagattg
     2461 tggaaatgat ggaatttttg atagaatatg gcccagcgca gctatattgg gaaccagctg
     2521 aagttttcct caaactttct tgtgtgcaac tcttgttgca gcttatttct attgcctgca
     2581 attggaagac ctattatgca aggaatgaca ctgtgcgctt tgctttggat gtcctggcta
     2641 ttcttactgt ggtgccaaaa atccagctcc agttggcaga atcagtggac gtgttggatg
     2701 aggctggatc tacagtctct actgtaggta tcagcattat tttgggagtg gctgagggtg
     2761 agttcttcat ccatgatgct gaaattcaga agtcagcact tcagattatc atcaattgtg
     2821 tgtgtggccc agataaccga atatccagta ttggtaaatt tatctctggt actcctcgga
     2881 gaaagctgcc tcagaaccct aaaagcagtg agcacaccct ggccaagatg tggaatgtgg
     2941 ttcagtccaa caacggcatc aaggtgctcc tgtccttact gtccattaag atgcccatca
     3001 cagatgcaga ccaaatccgg gccctggcct gcaaagccct agtgggcctg tctcgcagta
     3061 gcactgtccg gcagatcatc agtaaactgc cccttttcag cagctgccag atccagcagc
     3121 tgatgaagga gcctgtgctg caggacaagc gcagtgacca tgtcaagttc tgcaagtatg
     3181 ctgctgaact cattgaacgg gtgtcaggaa aaccacttct cattggcact gatgtttccc
     3241 tagcacgact gcagaaagca gatgttgttg cccagtcaag gatctccttc cctgagaaag
     3301 agctgctttt gttgatacga aaccatctta tttctaaagg gcttggagaa acagcaaccg
     3361 tgctgacaaa agaggctgac ctgcccatga ctgctgcctc ccattcttct gcctttaccc
     3421 cagtcactgc tgctgcttct cctgtctctc taccccgaac ccctcgtatc gctaatggca
     3481 ttgcaactcg tctgggcagc catgctgctg tgggtgcctc tgcgccttct gcccctactg
     3541 ctcatcctca gccacggccc ccccagggtc cgctagctct gcccggccca tcttatgcag
     3601 gcaactcccc tttgattggt agaatcagtt ttatcagaga gaggccatca ccctgcaatg
     3661 gcaggaaaat cagagtgttg cggcagaagt cggaccatgg tgcctacagc caaagcccag
     3721 ccataaaaaa acagctggac agacatcttc cttccccacc tacgctggac agtataatca
     3781 cagagtatct tagagaacaa catgctcgct gcaagaatcc agttgccacc tgcccacctt
     3841 tctccctctt tactcctcac caatgtcctg agccaaaaca gaggcggcaa gcgccaataa
     3901 actttacgtc aaggctaaac cgcagggcat catttccaaa gtatggaggg gtggatggcg
     3961 gatgctttga taggcacctt atctttagca gattccgtcc tatttcagtg ttccgggaag
     4021 ccaatgaaga tgagagtggc ttcacctgct gtgcattctc agcacgggag cggttcctga
     4081 tgcttggcac ctgcacaggg cagctgaagc tctataatgt gtttagtgga caggaggagg
     4141 ccagctataa ctgtcacaac tcagccatca cacatcttga accttccagg gatgggtcct
     4201 tgctgctgac atctgctact tggagccagc ctttgtctgc actttgggga atgaagtcag
     4261 tatttgatat gaagcattcc ttcacagaag atcactatgt tgagttcagt aagcactccc
     4321 aggatcgggt catcggcaca aaaggagaca ttgcccacat ttatgatatt cagactggca
     4381 acaagctgtt gactctgttt aacccagatc ttgccaacaa ctacaagagg aactgtgcca
     4441 cctttaatcc tacagatgat cttgtcttaa atgatggcgt cctctgggat gtccgctctg
     4501 cacaggccat ccacaagttt gacaagttca atatgaacat cagtggtgtt ttccatccaa
     4561 atggactgga ggtgatcatt aatactgaga tttgggacct tcgaactttt catcttttgc
     4621 atactgttcc cgctctggat cagtgtcgcg tggtgttcaa tcacacggga acagtgatgt
     4681 atggagctat gttgcaggca gatgatgaag atgacttaat ggaagagagg atgaaaagcc
     4741 cctttgggtc atccttccga acatttaatg caactgacta caaacctata gcaaccattg
     4801 atgtgaaacg gaacatcttt gacctgtgta cagacaccaa agactgctat cttgctgtca
     4861 ttgagaatca aggcagcatg gatgccctga acatggacac agtatgcagg ctgtatgaag
     4921 tgggcaggca gcgtctggca gaggatgagg atgaagagga ggaccaggaa gaggaagaac
     4981 aggaggaaga agatgatgat gaagatgatg atgacaccga tgatttagat gagcttgaca
     5041 ctgaccagtt gctggaggcg gagttggagg aggacgacaa taatgagaac gcaggggaag
     5101 atggggacaa tgacttctct ccctctgatg aggagctagc aaaccttcta gaggagggag
     5161 aggacgggga ggatgaagac tctgatgcag atgaggaggt ggaactgatc ctgggggaca
     5221 ctgacagctc tgacaactct gatttggaag atgacatcat cttatctctg aatgagggat
     5281 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     5341 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     5401 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5461 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5521 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5581 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5641 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5701 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5761 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5821 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5881 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     5941 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     6001 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     6061 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     6121 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     6181 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6241 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     6301 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6361 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6421 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6481 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6541 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6601 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     6661 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     6721 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     6781 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     6841 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     6901 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6961 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     7021 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     7081 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     7141 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     7201 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     7261 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     7321 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     7381 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     7441 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     7501 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     7561 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     7621 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     7681 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     7741 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     7801 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     7861 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     7921 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     7981 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     8041 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     8101 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     8161 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     8221 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     8281 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     8341 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     8401 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     8461 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     8521 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     8581 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     8641 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     8701 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     8761 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     8821 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     8881 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     8941 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     9001 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     9061 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     9121 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     9181 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     9241 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     9301 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     9361 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     9421 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     9481 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     9541 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     9601 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     9661 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     9721 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     9781 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     9841 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     9901 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     9961 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
    10021 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
    10081 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
    10141 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
    10201 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
    10261 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
    10321 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
    10381 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
    10441 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
    10501 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
    10561 ggcttgaccg acaattgcat gaagaatctg cttagg
//
