LOCUS       sequence-394905-        6462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid expresses delta WD40 deleted version of Myc-tagged FBW7 
            human isoform 1. WD40 All are deleted. 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 6462)
  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 6462)
  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 WD40 (All deleted)
FEATURES             Location/Qualifiers
     source          1..6462
                     /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(1954..1978)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2015..2033)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2016..2033)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2053..2070)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2059..2283
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2329..2757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2416..2435)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2626..2647
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2766..2786)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2771..3100
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2951..3086
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3013..3032
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3167..3961
                     /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(3221..3240)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3831..3850
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4135..4256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4172..4191)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4226..4245
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4305..4321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4305..4321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4318..4340)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4329..4345
                     /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(4353..4359,4360..4377,4378..4383))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4398..4419
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4536..4553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4707..5295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4787..4806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5466..6257,6258..6326))
                     /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     6089..6108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6327..6431)
                     /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 aaagaagagt gaaacccagc tttcttgtac aaagtggtga
     1981 taattcactc gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag
     2041 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2101 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2161 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2221 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     2281 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     2341 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2401 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2461 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2521 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2581 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2641 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2701 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     2761 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2821 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2881 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2941 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3001 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3061 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3121 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3181 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3241 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3301 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3361 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3421 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3481 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3541 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3601 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3661 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3721 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3781 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3841 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3901 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3961 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4021 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4081 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4141 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4201 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4261 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4321 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4381 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4441 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4501 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4561 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4621 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4681 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4741 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4801 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4861 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4921 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4981 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5041 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5101 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5161 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5221 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5281 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5341 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5401 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5461 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5521 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5581 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5641 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5701 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5761 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5821 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5881 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5941 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6001 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6061 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6121 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6181 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6241 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6301 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6361 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6421 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
