LOCUS       sequence-394916-        6738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid expresses delta WD40 deleted version of Myc-tagged FBW7 
            human isoform 1. WD40 propellars 2 to 7 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 6738)
  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 6738)
  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 WD40 (-2 to 7)
FEATURES             Location/Qualifiers
     source          1..6738
                     /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(2230..2254)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2291..2309)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2292..2309)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2329..2346)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2335..2559
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2605..3033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2692..2711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2902..2923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3042..3062)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3047..3376
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3227..3362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3289..3308
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3443..4237
                     /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(3497..3516)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4107..4126
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4411..4532
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4448..4467)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4502..4521
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4581..4597)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4581..4597)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4594..4616)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4605..4621
                     /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(4629..4635,4636..4653,4654..4659))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4674..4695
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4812..4829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4983..5571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5063..5082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5742..6533,6534..6602))
                     /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     6365..6384
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6603..6707)
                     /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 tttaaaagtt tggtcagcag tcacaggcaa atgtctgaga acattagtgg
     2221 gacattgaaa cccagctttc ttgtacaaag tggtgataat tcactcgagc atgcatctag
     2281 agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc
     2341 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2401 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2461 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2521 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2581 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2641 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2701 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2761 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2821 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2881 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2941 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3001 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3061 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3121 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3181 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3241 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3301 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3361 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     3421 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3481 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3541 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3601 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3661 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3721 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3781 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3841 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3901 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3961 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4021 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4081 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4141 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4201 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4261 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4321 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4381 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4441 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4501 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4561 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4621 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4681 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4741 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4801 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4861 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4921 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4981 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5041 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5101 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5161 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5221 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5281 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5341 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5401 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5461 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5521 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5581 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5641 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5701 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5761 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5821 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5881 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5941 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6001 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6061 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6121 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6181 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6241 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6301 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6361 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6421 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6481 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6541 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6601 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6661 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6721 aaagtgccac ctgacgtc
//
