LOCUS       sequence-394913-        7584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid expresses D-domain deleted version of Myc-tagged FBW7 
            human isoform 1. Used for mammalian overexpression and detection by 
            western blotting..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7584)
  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 7584)
  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 Delta D-Domain
FEATURES             Location/Qualifiers
     source          1..7584
                     /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(3076..3100)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(3137..3155)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3138..3155)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3175..3192)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3181..3405
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3451..3879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3538..3557)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3748..3769
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3888..3908)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3893..4222
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4073..4208
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4135..4154
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4289..5083
                     /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(4343..4362)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4953..4972
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5257..5378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5294..5313)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5348..5367
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5427..5443)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5427..5443)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5440..5462)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5451..5467
                     /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(5475..5481,5482..5499,5500..5505))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5520..5541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5658..5675)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5829..6417)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5909..5928)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6588..7379,7380..7448))
                     /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     7211..7230
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     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 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 aaatgggacc agagaaattg cttgctttag atgaactcat tgatagttgt gaaccaacac
     1741 aagtaaaaca tatgatgcaa gtgatagaac cccagtttca acgagacttc atttcattgc
     1801 tccctaaaga gttggcactc tatgtgcttt cattcctgga acccaaagac ctgctacaag
     1861 cagctcagac atgtcgctac tggagaattt tggctgaaga caaccttctc tggagagaga
     1921 aatgcaaaga agaggggatt gatgaaccat tgcacatcaa gagaagaaaa gtaataaaac
     1981 caggtttcat acacagtcca tggaaaagtg catacatcag acagcacaga attgatacta
     2041 actggaggcg aggagaactc aaatctccta aggtgctgaa aggacatgat gatcatgtga
     2101 tcacatgctt acagttttgt ggtaaccgaa tagttagtgg ttctgatgac aacactttaa
     2161 aagtttggtc agcagtcaca ggcaaatgtc tgagaacatt agtgggacat acaggtggag
     2221 tatggtcatc acaaatgaga gacaacatca tcattagtgg atctacagat cggacactca
     2281 aagtgtggaa tgcagagact ggagaatgta tacacacctt atatgggcat acttccactg
     2341 tgcgttgtat gcatcttcat gaaaaaagag ttgttagcgg ttctcgagat gccactctta
     2401 gggtttggga tattgagaca ggccagtgtt tacatgtttt gatgggtcat gttgcagcag
     2461 tccgctgtgt tcaatatgat ggcaggaggg ttgttagtgg agcatatgat tttatggtaa
     2521 aggtgtggga tccagagact gaaacctgtc tacacacgtt gcaggggcat actaatagag
     2581 tctattcatt acagtttgat ggtatccatg tggtgagtgg atctcttgat acatcaatcc
     2641 gtgtttggga tgtggagaca gggaattgca ttcacacgtt aacagggcac cagtcgttaa
     2701 caagtggaat ggaactcaaa gacaatattc ttgtctctgg gaatgcagat tctacagtta
     2761 aaatctggga tatcaaaaca ggacagtgtt tacaaacatt gcaaggtccc aacaagcatc
     2821 agagtgctgt gacctgttta cagttcaaca agaactttgt aattaccagc tcagatgatg
     2881 gaactgtaaa actatgggac ttgaaaacgg gtgaatttat tcgaaaccta gtcacattgg
     2941 agagtggggg gagtggggga gttgtgtggc ggatcagagc ctcaaacaca aagctggtgt
     3001 gtgcagttgg gagtcggaat gggactgaag aaaccaagct gctggtgctg gactttgatg
     3061 tggacatgaa gtgaaaccca gctttcttgt acaaagtggt gataattcac tcgagcatgc
     3121 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
     3181 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3241 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3301 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3361 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3421 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3481 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3541 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3601 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3661 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3721 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3781 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3841 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3901 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3961 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4021 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4081 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4141 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4201 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4261 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4321 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4381 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4441 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4501 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4561 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4621 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4681 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4741 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4801 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4861 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4921 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4981 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5041 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5101 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5161 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5221 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5281 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5341 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5401 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5461 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5521 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5581 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5641 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5701 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5761 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5821 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5881 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5941 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6001 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6061 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6121 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6181 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6241 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6301 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6361 agcggtggtt 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
//
