LOCUS       V046268                 9243 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046268
VERSION     V046268
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..3923
                     /label="Zfp451(NM_133817)"
                     /note="Zfp451(NM_133817)"
                     /gene="Zfp451"
     CDS             3930..3995
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4019..4607
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4673..4721
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4923..5351
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5365..5694
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5761..6555
                     /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    6731..6852
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6901..6917)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6925..6941
                     /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(6949..6979)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6994..7015
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7303..7891)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8062..8922)
                     /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(8923..9027)
                     /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 ccaccatggg agacccaggg ccggagataa
      781 tagaatctgt cccaccagct gggcccgagg catcagagtc taccacggat gaaaacgaag
      841 atgacattca gtttgttagt gaaggaccgt taagacccgt tttggaatac attgatctag
      901 tcagcagtga tgatgaagag cctagtacct ctcacagtga tgagaatttt aaatgcaaag
      961 actatataga tcaccagaag gataaagttg ctttaacttt ggctcgcctg gcccgccatg
     1021 ttgaagtaga aaaacaacag aaagaagaga agaacagggc atttagggaa aaaattgatt
     1081 ttcagcacgc tcatggttta caagagttgg agtttattca aggacattca gaaacagaag
     1141 cagcaagaca gtgtgtggac cagtggctaa aaatgccagg actcagaaca aatgcagcta
     1201 attctggaac aaaaagatca ttccaacgtg gaggcaggat gtggaggtct gagaagccaa
     1261 ttttgtgtcc tataatgcac tgtaacaagg aatttgacaa tgggcacctt ctgttgggac
     1321 atttgaaaag gtttgatcac tccccatgtg atcccacaat tacgttacat ggacctctgg
     1381 ctaattcgtt tgcatgtgca gtgtgctatg aacatttcgt tactcaacag cagtacaagg
     1441 accatcttct ctccaggaca gctgcagccg atggacatag caatagcctt cttcctcaga
     1501 ttattcaatg ttatgcttgt ccacagtgct tccttttgtt tagcaccaag gatgagtgtt
     1561 tgaagcatat gtctacaaag aatcatttcc atcagagctt taaactgagt gataataaag
     1621 gaacggcccg cccaatatca tttccatctt ttgcaaagaa gcgtttggtc tctctgtgca
     1681 aagatgttcc atttcaggtt aagtgtgtgg cctgccacca gactctgcgt tctcacatgg
     1741 agctcactgc ccatttcagg gttcgttgtc aaaatgctgg acctgttgct atagctgaaa
     1801 aaagcattac tcaggttgca aaagaattca tagtaagagg ttattgttca gattgcaacc
     1861 aggtctttat ggatgtagcc agcacccaaa gtcacaagaa ttcaggacac aaaattacac
     1921 ttgcgaactc ggtggaagaa tctgtcttgc tttattgcca catcagtgaa gggagtcggc
     1981 ctccttgtga tttacattta tttagtcaac caaaaatttc atcacttaaa cggattctgt
     2041 ccgttaaaga gtccagtgca gaggattgta tcgttccgac aaagaaggtg aacttaggtg
     2101 ttgaaagcct tggaggtgca actcgtgtgc agaggcagag tccagcagtc acagcctggt
     2161 tttgtgaatg cagacggcag tttcccagtg aagaggcggt agaaaagcat gttttctcgg
     2221 caaacacaat gtgttataag tgtgtggtct gtggaaaggt ttgtgaagat tcgggggtca
     2281 tgcgtttaca catgagccgg tttcatggag gggcgcattt aaataacttt ctattttggt
     2341 gtcggacgtg caaaaaggag ttagtaaaga aagatgccat catggcacac attactgagt
     2401 ttcatagtgg gcatagatat ttttatgaga tggatgaggt agaggaggaa gaggaggagg
     2461 ccatgccgtc atcctctgtg gagagccatt tgaatactga caaacctcct tcacccattg
     2521 ctgttgttga tcactgcccg gccaacagtc ctcccagggg caggtggcag tgccgcatct
     2581 gtgaggacat gtttgaatcc caggagtgtg tgaagcagca ctgcatgtcc ttgacaagcc
     2641 accggtttca cagatacagc tgtgcccact gcagaaagac tttccataaa gtggaaactc
     2701 tgtaccgaca ttgccaggac gagcatgaca gtgagatcat gatgaagtac ttctgtgggc
     2761 tttgtgatct catttttaac aaggaagaag aatttctgag tcactataag gaacaccaca
     2821 gcatagatta tgtgtttgtg tcagaaaaaa ctaaaacctc aattaaaact gaaggcgatt
     2881 ttaaaatagt agagaccagt agtttactaa gctgtggctg tcatgagagt tacatgtgta
     2941 aaatcaacag aaaggaagac tatgaccgat gccttccagt cctgctggag aaaggtaggc
     3001 tgtggtttcg ctgcagctcg tgctcagcca cagcacagaa tgtgaccgac attaacactc
     3061 acgtctgcca agtgcacaga aaagagaaaa gtgaggagga gcagcagtat gtgatcaagt
     3121 gtggcatctg caccaaggca ttccagaaca cggagagcgc tcagcagcac ttccacagga
     3181 agcacgcggc cctccagaaa cccaccgcga ccccaggggg agccaacaga agcagcacat
     3241 gccagctggc tgctagtgcc tcacatgctg agaaaaacct gaaacagcct agctctcaga
     3301 aacattcaga cgtggaaaaa ggagctgagc atgatgtacg ctgccagaac atagaggagg
     3361 aagttgagct tccggatgtg gactacctgc ggaccatgac tcacatagtc tttgtagatt
     3421 ttgataactg gtccaacttt tttggtcatc tacctgggca tcttaatcaa ggaacgttta
     3481 tttggggctt tcaaggggga aacaccaact ggaagccccc gctcagctgt aaggtctata
     3541 attacttgag caggattggc tgcttcttcc ttcatcctcg ctgcagtaaa agaaaagatg
     3601 ccgcggattt tgccatatgt atgcatgctg gccgtctaga tgagcaactt cccaaacaaa
     3661 ttcctttcac catcctctcg ggagaccaag gctttctgga gctagagaat caatttaaga
     3721 agacccagag gccagctcat atactaaacc ctcaccactt agagggagac atgatgtgtg
     3781 ccttgttaaa tagcatatct gatactacca aagagtgtga cagtgacgat agctcgggga
     3841 tgaaaggatc tccagcagag gagctcaggg ccacggagga tgtggaatta gaagaagcta
     3901 ttagaagaag tcttgaggag atgggatccg actacaaaga ccatgacggt gattataaag
     3961 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     4021 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4081 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4141 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4201 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4261 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4321 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4381 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     4441 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4501 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     4561 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     4621 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4681 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4741 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4801 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4861 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     4921 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4981 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5041 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     5101 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5161 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5221 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5281 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5341 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5401 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5461 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5521 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5581 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5641 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5701 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5761 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5821 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5881 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5941 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6001 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6061 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6121 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6181 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6241 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6301 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6361 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6421 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6481 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6541 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     6601 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6661 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6721 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6781 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6841 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6901 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6961 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7021 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7081 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7141 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7201 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7261 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7321 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7381 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7441 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     7501 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7561 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7621 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7681 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7741 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7801 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7861 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7921 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7981 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8041 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8101 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8161 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8221 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8281 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8341 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8401 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8461 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8521 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8581 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8641 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8701 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8761 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8821 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8881 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8941 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9001 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     9061 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     9121 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     9181 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     9241 agg
//
