LOCUS       V037478                 6703 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037478
VERSION     V037478
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..2223
                     /label="MBD2(NM_003927)"
                     /note="MBD2(NM_003927)"
                     /gene="MBD2"
     polyA_signal    2303..2527
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2573..3001
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3015..3344
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3411..4205
                     /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    4379..4500
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4549..4565)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4573..4589
                     /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(4597..4627)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4642..4663
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4951..5536)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5707..6567)
                     /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(6568..6672)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atgcgcgcgc acccgggggg aggccgctgc
     1021 tgcccggagc aggaggaggg ggagagtgcg gcgggcggca gcggcgctgg cggcgactcc
     1081 gccatagagc aggggggcca gggcagcgcg ctcgccccgt ccccggtgag cggcgtgcgc
     1141 agggaaggcg ctcggggcgg cggccgtggc cgggggcggt ggaagcaggc gggccggggc
     1201 ggcggcgtct gtggccgtgg ccggggccgg ggccgtggcc ggggacgggg acggggccgg
     1261 ggccggggcc gcggccgtcc cccgagtggc ggcagcggcc ttggcggcga cggcggcggc
     1321 tgcggcggcg gcggcagcgg tggcggcggc gccccccggc gggagccggt ccctttcccg
     1381 tcggggagcg cggggccggg gcccagggga ccccgggcca cggagagcgg gaagaggatg
     1441 gattgcccgg ccctcccccc cggatggaag aaggaggaag tgatccgaaa atctgggcta
     1501 agtgctggca agagcgatgt ctactacttc agtccaagtg gtaagaagtt cagaagcaag
     1561 cctcagttgg caaggtacct gggaaatact gttgatctca gcagttttga cttcagaact
     1621 ggaaagatga tgcctagtaa attacagaag aacaaacaga gactgcgaaa cgatcctctc
     1681 aatcaaaata agggtaaacc agacttgaat acaacattgc caattagaca aacagcatca
     1741 attttcaaac aaccggtaac caaagtcaca aatcatccta gtaataaagt gaaatcagac
     1801 ccacaacgaa tgaatgaaca gccacgtcag cttttctggg agaagaggct acaaggactt
     1861 agtgcatcag atgtaacaga acaaattata aaaaccatgg aactacccaa aggtcttcaa
     1921 ggagttggtc caggtagcaa tgatgagacc cttttatctg ctgttgccag tgctttgcac
     1981 acaagctctg cgccaatcac agggcaagtc tccgctgctg tggaaaagaa ccctgctgtt
     2041 tggcttaaca catctcaacc cctctgcaaa gcttttattg tcacagatga agacatcagg
     2101 aaacaggaag agcgagtaca gcaagtacgc aagaaattgg aagaagcact gatggcagac
     2161 atcttgtcgc gagctgctga tacagaagag atggatattg aaatggacag tggagatgaa
     2221 gcctaagaat tctgcagata tccagcacag tggcggccgc tcgagtctag agggcccgtt
     2281 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2341 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2401 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2461 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2521 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2581 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2641 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2701 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2761 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2821 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2881 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2941 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3001 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3061 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3181 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3241 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3301 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3361 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3421 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3481 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3541 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3601 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3661 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3721 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3781 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3841 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3901 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3961 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4021 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4081 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4141 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4201 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4261 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4321 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4381 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4441 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4501 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4561 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4621 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4681 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4741 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4801 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4861 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4921 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4981 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5041 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5101 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5161 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5221 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5281 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5341 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5401 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5461 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     5521 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5581 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5641 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5701 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5761 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5821 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5881 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5941 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6001 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6061 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6121 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6181 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6241 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6301 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6361 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6421 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6481 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6541 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6601 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6661 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
