LOCUS       sequence-394919-        7599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid expresses Myc-tagged FBW7 human isoform 1 with Lys404 to
            Arg mutation. Used for overexpression and detection by western 
            blotting..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7599)
  AUTHORS   Khan OM, Almagro J, Nelson JK, Horswell S, Encheva V, Keyan KS, 
            Clurman BE, Snijders AP, Behrens A
  TITLE     Proteasomal degradation of the tumour suppressor FBW7 requires 
            branched ubiquitylation by TRIP12.
  JOURNAL   Nat Commun. 2021 Apr 6;12(1):2043. doi: 10.1038/s41467-021-22319-5.
  PUBMED    33824312
REFERENCE   2  (bases 1 to 7599)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2021-04-6"; volume: "12(1):2043. doi"; pages: " 
            10.1038/s41467-021-22319-5"
COMMENT     Sequence Label: pDEST-Myc-FBW7]K404R
FEATURES             Location/Qualifiers
     source          1..7599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             903..932
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    942..966
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3091..3115)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(3152..3170)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3153..3170)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3190..3207)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3196..3420
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3466..3894
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3553..3572)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3763..3784
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3903..3923)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3908..4237
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4088..4223
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4150..4169
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4304..5098
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4358..4377)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4968..4987
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5272..5393
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5309..5328)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5363..5382
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5442..5458)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5442..5458)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5455..5477)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5466..5482
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5490..5496,5497..5514,5515..5520))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5535..5556
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5673..5690)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5844..6432)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5924..5943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6603..7394,7395..7463))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7226..7245
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7464..7568)
                     /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 gcttccacca
      901 tggaacaaaa gctgatttct gaagaagact tgggaattat cacaagtttg tacaaaaaag
      961 caggctttaa tcaggaactg ctctctgtgg gcagcaaaag acgacgaact ggaggctctc
     1021 tgagaggtaa cccttcctca agccaggtag atgaagaaca gatgaatcgt gtggtagagg
     1081 aggaacagca acagcaactc agacaacaag aggaggagca cactgcaagg aatggtgaag
     1141 ttgttggagt agaacctaga cctggaggcc aaaatgattc ccagcaagga cagttggaag
     1201 aaaacaataa tagatttatt tcggtagatg aggactcctc aggaaaccaa gaagaacaag
     1261 aggaagatga agaacatgct ggtgaacaag atgaggagga tgaggaggag gaggagatgg
     1321 accaggagag tgacgatttt gatcagtctg atgatagtag cagagaagat gaacatacac
     1381 atactaacag tgtcacgaac tccagtagta ttgtggacct gcccgttcac caactctcct
     1441 ccccattcta tacaaaaaca acaaaaatga aaagaaagtt ggaccatggt tctgaggtcc
     1501 gctctttttc tttgggaaag aaaccatgca aagtctcaga atatacaagt accactgggc
     1561 ttgtaccatg ttcagcaaca ccaacaactt ttggggacct cagagcagcc aatggccaag
     1621 ggcaacaacg acgccgaatt acatctgtcc agccacctac aggcctccag gaatggctaa
     1681 aaatgtttca gagctggagt ggaccagaga aattgcttgc tttagatgaa ctcattgata
     1741 gttgtgaacc aacacaagta aaacatatga tgcaagtgat agaaccccag tttcaacgag
     1801 acttcatttc attgctccct aaagagttgg cactctatgt gctttcattc ctggaaccca
     1861 aagacctgct acaagcagct cagacatgtc gctactggag aattttggct gaagacaacc
     1921 ttctctggag agagaaatgc aaagaagagg ggattgatga accattgcac atcaagagaa
     1981 gaaaagtaat aaaaccaggt ttcatacaca gtccatggaa aagtgcatac atcagacagc
     2041 acagaattga tactaactgg aggcgaggag aactcaaatc tcctaaggtg ctgaaaggac
     2101 atgatgatca tgtgatcaca tgcttacagt tttgtggtaa ccgaatagtt agtggttctg
     2161 atgacaacac tttaagagtt tggtcagcag tcacaggcaa atgtctgaga acattagtgg
     2221 gacatacagg tggagtatgg tcatcacaaa tgagagacaa catcatcatt agtggatcta
     2281 cagatcggac actcaaagtg tggaatgcag agactggaga atgtatacac accttatatg
     2341 ggcatacttc cactgtgcgt tgtatgcatc ttcatgaaaa aagagttgtt agcggttctc
     2401 gagatgccac tcttagggtt tgggatattg agacaggcca gtgtttacat gttttgatgg
     2461 gtcatgttgc agcagtccgc tgtgttcaat atgatggcag gagggttgtt agtggagcat
     2521 atgattttat ggtaaaggtg tgggatccag agactgaaac ctgtctacac acgttgcagg
     2581 ggcatactaa tagagtctat tcattacagt ttgatggtat ccatgtggtg agtggatctc
     2641 ttgatacatc aatccgtgtt tgggatgtgg agacagggaa ttgcattcac acgttaacag
     2701 ggcaccagtc gttaacaagt ggaatggaac tcaaagacaa tattcttgtc tctgggaatg
     2761 cagattctac agttaaaatc tgggatatca aaacaggaca gtgtttacaa acattgcaag
     2821 gtcccaacaa gcatcagagt actgtgacct gtttacagtt caacaagaac tttgtaatta
     2881 ccagctcaga tgatggaact gtaaaactat gggacttgaa aacgggtgaa tttattcgaa
     2941 acctagtcac attggagagt ggggggagtg ggggagttgt gtggcggatc agagcctcaa
     3001 acacaaagct ggtgtgtgca gttgggagtc ggaatgggac tgaagaaacc aagctgctgg
     3061 tgctggactt tgatgtggac atgaagtgaa acccagcttt cttgtacaaa gtggtgataa
     3121 ttcactcgag catgcatcta gagggcccta ttctatagtg tcacctaaat gctagagctc
     3181 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3241 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3301 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3361 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     3421 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3481 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3541 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3601 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3661 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3721 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3781 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3841 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     3901 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3961 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4021 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4081 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4141 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     4201 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4261 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4321 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4381 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4441 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4501 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4561 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4621 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4681 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4741 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4801 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4861 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4921 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4981 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5041 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5101 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5161 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5221 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5281 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5341 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5401 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5461 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5521 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5581 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5641 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5701 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5761 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5821 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5881 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5941 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6001 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6061 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6121 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6181 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6241 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6301 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6361 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6421 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6481 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6541 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6601 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6661 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6721 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6781 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6841 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6901 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6961 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7021 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7081 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7141 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7201 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7261 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7321 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7381 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7441 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7501 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7561 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
