LOCUS       V047414                 7584 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047414
VERSION     V047414
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"
     CDS             905..3025
                     /label="FBXW7(NM_033632)"
                     /note="FBXW7(NM_033632)"
                     /gene="FBXW7"
     CDS             3041..3082
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     polyA_signal    3184..3408
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3454..3882
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3896..4225
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4292..5086
                     /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    5260..5381
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5430..5446)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5454..5470
                     /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(5478..5508)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5523..5544
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5832..6417)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6588..7448)
                     /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(7449..7553)
                     /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 caccatgaat caggaactgc tctctgtggg cagcaaaaga cgacgaactg gaggctctct
      961 gagaggtaac ccttcctcaa gccaggtaga tgaagaacag atgaatcgtg tggtagagga
     1021 ggaacagcaa cagcaactca gacaacaaga ggaggagcac actgcaagga atggtgaagt
     1081 tgttggagta gaacctagac ctggaggcca aaatgattcc cagcaaggac agttggaaga
     1141 aaacaataat agatttattt cggtagatga ggactcctca ggaaaccaag aagaacaaga
     1201 ggaagatgaa gaacatgctg gtgaacaaga tgaggaggat gaggaggagg aggagatgga
     1261 ccaggagagt gacgattttg atcagtctga tgatagtagc agagaagatg aacatacaca
     1321 tactaacagt gtcacgaact ccagtagtat tgtggacctg cccgttcacc aactctcctc
     1381 cccattctat acaaaaacaa caaaaatgaa aagaaagttg gaccatggtt ctgaggtccg
     1441 ctctttttct ttgggaaaga aaccatgcaa agtctcagaa tatacaagta ccactgggct
     1501 tgtaccatgt tcagcaacac caacaacttt tggggacctc agagcagcca atggccaagg
     1561 gcaacaacga cgccgaatta catctgtcca gccacctaca ggcctccagg aatggctaaa
     1621 aatgtttcag agctggagtg gaccagagaa attgcttgct ttagatgaac tcattgatag
     1681 ttgtgaacca acacaagtaa aacatatgat gcaagtgata gaaccccagt ttcaacgaga
     1741 cttcatttca ttgctcccta aagagttggc actctatgtg ctttcattcc tggaacccaa
     1801 agacctgcta caagcagctc agacatgtcg ctactggaga attttggctg aagacaacct
     1861 tctctggaga gagaaatgca aagaagaggg gattgatgaa ccattgcaca tcaagagaag
     1921 aaaagtaata aaaccaggtt tcatacacag tccatggaaa agtgcataca tcagacagca
     1981 cagaattgat actaactgga ggcgaggaga actcaaatct cctaaggtgc tgaaaggaca
     2041 tgatgatcat gtgatcacat gcttacagtt ttgtggtaac cgaatagtta gtggttctga
     2101 tgacaacact ttaaaagttt ggtcagcagt cacaggcaaa tgtctgagaa cattagtggg
     2161 acatacaggt ggagtatggt catcacaaat gagagacaac atcatcatta gtggatctac
     2221 agatcggaca ctcaaagtgt ggaatgcaga gactggagaa tgtatacaca ccttatatgg
     2281 gcatacttcc actgtgcgtt gtatgcatct tcatgaaaaa agagttgtta gcggttctcg
     2341 agatgccact cttagggttt gggatattga gacaggccag tgtttacatg ttttgatggg
     2401 tcatgttgca gcagtccgct gtgttcaata tgatggcagg agggttgtta gtggagcata
     2461 tgattttatg gtaaaggtgt gggatccaga gactgaaacc tgtctacaca cgttgcaggg
     2521 gcatactaat agagtctatt cattacagtt tgatggtatc catgtggtga gtggatctct
     2581 tgatacatca atccgtgttt gggatgtgga gacagggaat tgcattcaca cgttaacagg
     2641 gcaccagtcg ttaacaagtg gaatggaact caaagacaat attcttgtct ctgggaatgc
     2701 agattctaca gttaaaatct gggatatcaa aacaggacag tgtttacaaa cattgcaagg
     2761 tcccaacaag catcagagtg ctgtgacctg tttacagttc aacaagaact ttgtaattac
     2821 cagctcagat gatggaactg taaaactatg ggacttgaaa acgggtgaat ttattcgaaa
     2881 cctagtcaca ttggagagtg gggggagtgg gggagttgtg tggcggatca gagcctcaaa
     2941 cacaaagctg gtgtgtgcag ttgggagtcg gaatgggact gaagaaacca agctgctggt
     3001 gctggacttt gatgtggaca tgaagggtgg aggcggttca ggtaagccta tccctaaccc
     3061 tctcctcggt ctcgattcta cataggatcc actagtccag tgtggtggaa ttctgcagat
     3121 atccagcaca gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct
     3181 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3241 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3301 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3361 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3421 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3481 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3541 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3601 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3661 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3721 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3781 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3841 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3901 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3961 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4021 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4081 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4141 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     4201 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4261 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4321 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4381 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4441 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4501 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4561 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4621 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4681 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4741 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4801 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4861 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4921 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4981 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5041 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5101 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5161 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5221 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5281 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5341 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5401 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5461 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5521 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5581 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5641 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5701 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5761 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5821 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5881 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5941 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6001 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6061 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6121 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6181 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6241 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6301 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6361 ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6421 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6481 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6541 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6601 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6661 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6721 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6781 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6841 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6901 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6961 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7021 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7081 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7141 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7201 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7261 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7321 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7381 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7441 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7501 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7561 ccccgaaaag tgccacctga cgtc
//
