LOCUS       V046228                 7338 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046228
VERSION     V046228
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..2018
                     /label="Zfp57(NM_001013745)"
                     /note="Zfp57(NM_001013745)"
                     /gene="Zfp57"
     CDS             2025..2090
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2114..2702
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2768..2816
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3018..3446
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3460..3789
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3856..4650
                     /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    4826..4947
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4996..5012)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5020..5036
                     /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(5044..5074)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5089..5110
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5398..5986)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6157..7017)
                     /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(7018..7122)
                     /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 ccaccatggc agctaggaaa cagtcttccc
      781 agccatccag gacaccagtc agttatgagg acgtggcagt gtctttcacc caggaagaat
      841 gggaatatct tacttctaca cagaagaccc tttaccagaa agtgatgtca gaaaccttca
      901 agaacctgac atttgtcgga agcaagaaga aacctcaaga acctagctca gatctgcaag
      961 ataagaacga ggagcaggag aagtcctcca gttgcacagg ggtattcaaa ggtggaccat
     1021 tctttttctg tctgacctgt ggcaaatgtt tcaaaaagaa caccttcctc tttaatcacc
     1081 agtttcctgt gaggtcccgg aggctggcag tcacaaatcc acaaagccgc aaaggcaagg
     1141 gctacaaggc tcagcatcgt ggagagaggc ctttcttttg taatttctgt ggcaagactt
     1201 accgtgatgc ttctggactg agccgtcacc gacgtgctca tttaggttat aggccccgtt
     1261 catgccctga gtgtggaaag tgtttccggg atcagtctga ggtcaaccgt cacctgaagg
     1321 tacaccaaaa caagccagca gctagcaacc aggctggcaa ccaggctagc aaccagaggc
     1381 tgaagagtag ggttccacct acaacaccta gatcccaagc gcccgccctc aagtatgtga
     1441 aagtgatcca gggaccagtg gccagggcta aggcacggaa cagcggagcc tcgaccctga
     1501 atgtcagatc caactctatt acagtggtcc gttcaagaga aaagatctct tgtccctatt
     1561 gtcacataac gtttaccatg agaacctgtc tcttaaccca cctcaagatc cacttcagac
     1621 gtcaacccaa ccagcacttc tgctgcaaag agtcggccca ctcatccaac acactcagaa
     1681 tgcaaaagat ctacacttgc cccgtctgtg acagctcctt taggggaaag gagagcctgc
     1741 tggatcactt gtgctgccaa agaccaatca gattcagtaa atgctgggaa atcctgggtc
     1801 atttgctcgg ctatcttcat gaacccgtgg tgctgggaaa tatttttaaa gtaagggact
     1861 cctcgggaaa gaggatggaa tccaggagga gaagacggaa acgtgcctgc actgagaatc
     1921 ctgaaacaga aggcctgtct gggaaaggta gggtggctcc gtgggaaatg gagggtgcca
     1981 ccagccctga gagtcctgtg acagaagaag attcggacgg atccgactac aaagaccatg
     2041 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     2101 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2161 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2221 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2281 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2341 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2401 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2461 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     2521 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     2581 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     2641 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     2701 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     2761 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     2821 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     2881 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     2941 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     3001 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3061 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3121 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     3181 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3241 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3301 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3361 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     3421 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3481 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3541 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3601 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3661 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3721 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3781 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3841 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3901 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3961 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4021 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4081 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4141 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4201 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4261 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4321 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4381 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4441 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4501 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4561 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4621 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     4681 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4741 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4801 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4861 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4921 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4981 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5041 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5101 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5161 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5221 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5281 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5341 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5401 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5461 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5521 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5581 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5641 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5701 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5761 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5821 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5881 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5941 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6001 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6061 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6121 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6181 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6241 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6301 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6361 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6421 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6481 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6541 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6601 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6661 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6721 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6781 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6841 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6901 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6961 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7021 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7081 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7141 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     7201 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     7261 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     7321 atgaagaatc tgcttagg
//
