LOCUS       V018825                 7243 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018825
VERSION     V018825
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             997..2763
                     /label="FBXW7(NM_001013415)"
                     /note="FBXW7(NM_001013415)"
                     /gene="FBXW7"
     polyA_signal    2843..3067
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3113..3541
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3555..3884
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3951..4745
                     /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    4919..5040
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5089..5105)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5113..5129
                     /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(5137..5167)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5182..5203
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5491..6076)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6247..7107)
                     /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(7108..7212)
                     /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 ggtaccatgt caaaaccggg aaaacctact
     1021 ctaaaccatg gcttggttcc tgttgatctt aaaagtgcaa aagagcctct accacatcaa
     1081 actgtgatga agatatttag cattagcatc attgcccaag gcctcccttt ttgtcgaaga
     1141 cggatgaaaa gaaagttgga ccatggttct gaggtccgct ctttttcttt gggaaagaaa
     1201 ccatgcaaag tctcagaata tacaagtacc actgggcttg taccatgttc agcaacacca
     1261 acaacttttg gggacctcag agcagccaat ggccaagggc aacaacgacg ccgaattaca
     1321 tctgtccagc cacctacagg cctccaggaa tggctaaaaa tgtttcagag ctggagtgga
     1381 ccagagaaat tgcttgcttt agatgaactc attgatagtt gtgaaccaac acaagtaaaa
     1441 catatgatgc aagtgataga accccagttt caacgagact tcatttcatt gctccctaaa
     1501 gagttggcac tctatgtgct ttcattcctg gaacccaaag acctgctaca agcagctcag
     1561 acatgtcgct actggagaat tttggctgaa gacaaccttc tctggagaga gaaatgcaaa
     1621 gaagagggga ttgatgaacc attgcacatc aagagaagaa aagtaataaa accaggtttc
     1681 atacacagtc catggaaaag tgcatacatc agacagcaca gaattgatac taactggagg
     1741 cgaggagaac tcaaatctcc taaggtgctg aaaggacatg atgatcatgt gatcacatgc
     1801 ttacagtttt gtggtaaccg aatagttagt ggttctgatg acaacacttt aaaagtttgg
     1861 tcagcagtca caggcaaatg tctgagaaca ttagtgggac atacaggtgg agtatggtca
     1921 tcacaaatga gagacaacat catcattagt ggatctacag atcggacact caaagtgtgg
     1981 aatgcagaga ctggagaatg tatacacacc ttatatgggc atacttccac tgtgcgttgt
     2041 atgcatcttc atgaaaaaag agttgttagc ggttctcgag atgccactct tagggtttgg
     2101 gatattgaga caggccagtg tttacatgtt ttgatgggtc atgttgcagc agtccgctgt
     2161 gttcaatatg atggcaggag ggttgttagt ggagcatatg attttatggt aaaggtgtgg
     2221 gatccagaga ctgaaacctg tctacacacg ttgcaggggc atactaatag agtctattca
     2281 ttacagtttg atggtatcca tgtggtgagt ggatctcttg atacatcaat ccgtgtttgg
     2341 gatgtggaga cagggaattg cattcacacg ttaacagggc accagtcgtt aacaagtgga
     2401 atggaactca aagacaatat tcttgtctct gggaatgcag attctacagt taaaatctgg
     2461 gatatcaaaa caggacagtg tttacaaaca ttgcaaggtc ccaacaagca tcagagtgct
     2521 gtgacctgtt tacagttcaa caagaacttt gtaattacca gctcagatga tggaactgta
     2581 aaactatggg acttgaaaac gggtgaattt attcgaaacc tagtcacatt ggagagtggg
     2641 gggagtgggg gagttgtgtg gcggatcaga gcctcaaaca caaagctggt gtgtgcagtt
     2701 gggagtcgga atgggactga agaaaccaag ctgctggtgc tggactttga tgtggacatg
     2761 aagtgagaat tctgcagata tccagcacag tggcggccgc tcgagtctag agggcccgtt
     2821 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2881 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2941 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3001 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3061 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3121 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3181 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3241 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3301 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3361 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3421 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3481 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3541 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3601 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3661 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3721 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3781 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3841 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3901 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3961 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4021 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4081 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4141 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4201 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4261 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4321 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4381 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4441 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4501 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4561 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4621 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4681 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4741 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4801 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4861 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4921 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4981 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5041 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5101 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5161 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5221 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5281 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5341 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5401 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5461 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5521 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5581 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5641 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5701 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5761 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5821 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5881 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5941 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6001 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     6061 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6121 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6181 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6241 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6301 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6361 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6421 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6481 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6541 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6601 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6661 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6721 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6781 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6841 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6901 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6961 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7021 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7081 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7141 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7201 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
