LOCUS       V046113                 7107 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046113
VERSION     V046113
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..1787
                     /label="Dffb(NM_007859)"
                     /note="Dffb(NM_007859)"
                     /gene="Dffb"
     CDS             1794..1859
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1883..2471
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2537..2585
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2787..3215
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3229..3558
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3625..4419
                     /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    4595..4716
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4765..4781)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4789..4805
                     /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(4813..4843)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4858..4879
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5167..5755)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5926..6786)
                     /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(6787..6891)
                     /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 ccaccatgtg cgcggtgctc cgccaaccca
      781 aatgcgtcaa gttgcgagcc ctacatagcg cctgcaagtt cggcgtggcg gcccggagct
      841 gccaggagct gctgcgtaag ggctgcgtcc gcttccagct cccgatgccc ggttcccggc
      901 tgtgcctgta cgaagatggc acggaggtga cggacgactg cttcccgggc cttcccaacg
      961 acgctgagct cctattgctc accgctggcg agacctggca tggctatgtg agtgacatca
     1021 cacgtttcct cagtgtgttt aatgagccac atgccggcgt catccaggct gcacggcaac
     1081 tgctgtcaga tgagcaggcc ccactgaggc aaaagctgct ggccgatctt ctgcatcacg
     1141 tgagccagaa tattactgca gagacccggg agcaggaccc atcctggttt gaaggtttgg
     1201 agtcgagatt caggaataag tcgggctatc tgagatacag ctgtgagagt cggatccggg
     1261 gttacctaag agaggtgagc gcttacacct ctatggtgga tgaagcagct caagaagagt
     1321 acctgcgagt ccttggctcc atgtgccaga agctcaaatc ggtgcagtac aatggcagct
     1381 atttcgacag aggtgcagag gccagcagcc gcctctgtac tccagaagga tggttctcct
     1441 gccagggccc ctttgacctg gagagctgtc tttccaagca ctccatcaac ccctatggca
     1501 acagagagag ccggatcctc ttcagtacct ggaacctgga tcatataata gagaagaagc
     1561 gcaccgtggt acccacgctg gctgaagcca tccaggatgg gagggaggtg aactgggagt
     1621 acttctacag cctgctcttc actgccgaga acctgaagct ggtgcacatc gcctgccaca
     1681 agaagaccac acacaagctg gagtgcgacc gcagtaggat ctatcggcct cagacaggat
     1741 ccaggaggaa gcagcctgct cggaagaagc gccctgctcg gaagcgcgga tccgactaca
     1801 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     1861 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     1921 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1981 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2041 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2101 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2161 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2221 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     2281 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2341 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2401 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2461 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     2521 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     2581 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     2641 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     2701 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     2761 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2821 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2881 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2941 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3001 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3061 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3121 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     3181 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3241 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3301 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3361 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3421 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3481 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3541 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3601 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3661 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3721 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3781 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3841 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3901 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3961 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4021 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4081 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4141 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4201 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4261 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4321 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4381 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     4441 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4501 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4561 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4621 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4681 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4741 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4801 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4861 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4921 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4981 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5041 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5101 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5161 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5221 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5281 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5341 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5401 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5461 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5521 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5581 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5641 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5701 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5761 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5821 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5881 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5941 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6001 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6061 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6121 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6181 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6241 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6301 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6361 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6421 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6481 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6541 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6601 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6661 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6721 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6781 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6841 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6901 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     6961 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     7021 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     7081 gacaattgca tgaagaatct gcttagg
//
