LOCUS       sequence-425856-        6266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PPWD1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6266)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova YA, Baghaie AJ, Beldar S, Bolotokova A, Centrella
            PA, Chau I, Clark MA, Cuozzo JW, Dehghani-Tafti S, Disch JS, Dong A,
            Dumas A, Feng JA, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green SR,
            Guie MA, Guilinger JP, Harms N, Herasymenko O, Houliston S, 
            Hutchinson A, Kearnes S, Keefe AD, Kimani SW, Kramer T, Kutera M, 
            Kwak HA, Lento C, Li Y, Liu J, Loup J, Machado RAC, Mulhern CJ, 
            Perveen S, Righetto GL, Riley P, Shrestha S, Sigel EA, Silva M, 
            Sintchak MD, Slakman BL, Taylor RD, Thompson J, Torng W, 
            Underkoffler C, von Rechenberg M, Walsh RT, Watson I, Wilson DJ, 
            Wolf E, Yadav M, Yazdi AK, Zhang J, Zhang Y, Santhakumar V, Edwards 
            AM, Barsyte-Lovejoy D, Schapira M, Brown PJ, Halabelian L, 
            Arrowsmith CH
  TITLE     A Target Class Ligandability Evaluation of WD40 Repeat-Containing 
            Proteins.
  JOURNAL   J Med Chem. 2025 Jan 23;68(2):1092-1112. doi: 
            10.1021/acs.jmedchem.4c02010. Epub 2024 Nov 4.
  PUBMED    39495097
REFERENCE   2  (bases 1 to 6266)
  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"; doi: 
            "10.1021/acs.jmedchem.4c02010"; journalName: "J Med Chem"; date: 
            "2025-01-23- 23"; volume: "68"; issue: "2"; pages: "1092-1112"
COMMENT     Sequence Label: pBiT3.1-C_PPWD1:1-646
FEATURES             Location/Qualifiers
     source          1..6266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1241..1250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3329..3450
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3340..3359)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3565..3598)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3608..3655
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3687..4481
                     /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(3741..3760)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4351..4370
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4639..5227)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4719..4738)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5353..5401
                     /note="synthetic polyadenylation signal"
     CDS             complement(5425..5823)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5820..5829
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5854..6211)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5868..6003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6020..6211
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6168..6188
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggcga tcgcaacaag cttcgggaag
     1201 atctggagct agcggcgggg gatccggtgg gaattcgaca ccatggcggc ggaaagtggt
     1261 agcgattttc agcagagacg tagaaggcgc cgggacccgg aggaaccgga aaaaacagaa
     1321 ctcagcgaaa gagagctggc agtagcagtg gcggtgtccc aggagaacga tgaggagaac
     1381 gaagagcgct gggttggacc tttacctgtg gaggcaacac tggccaagaa gaggaaagtc
     1441 ttagagtttg aaagagtcta tcttgataat ctccccagtg catccatgta tgagcgcagt
     1501 tacatgcata gagatgttat cacccatgtg gtatgcacca aaacagattt tattattact
     1561 gccagtcatg atggacatgt caagttctgg aaaaaaatag aagagggaat tgaatttgtt
     1621 aaacattttc gtagtcacct gggagttatt gagagtattg cagttagctc tgagggagca
     1681 ttgttctgtt ctgtgggtga tgataaagca atgaaggtgt ttgatgtagt gaactttgac
     1741 atgatcaaca tgctgaaact tggctatttt cctggacagt gtgagtggat ctattgccca
     1801 ggggatgcaa tttcttcagt tgctgcttcc gaaaagagta caggaaaaat tttcatttat
     1861 gatggccgag gagataacca gccacttcat atttttgaca aactccatac atcacctctt
     1921 actcagatac ggctaaaccc agtttacaaa gcagtagtgt cttctgacaa atctgggatg
     1981 attgaatact ggactgggcc tcctcatgaa tataaattcc ccaaaaatgt gaactgggaa
     2041 tataaaactg acactgattt atatgaattt gccaagtgta aggcttatcc aaccagcgta
     2101 tgtttttcac cagatgggaa gaaaatagct actattggtt ctgatagaaa agttagaatt
     2161 ttcagatttg taactggaaa actcatgaga gtctttgatg aatcactaag catgtttact
     2221 gaactgcaac agatgaggca acagttacca gacatggaat ttggccgacg aatggctgta
     2281 gaacgtgagt tggagaaggt tgatgctgta agattaatta atatagtttt tgatgaaact
     2341 ggacacttcg tgctgtatgg aacaatgctg ggcattaaag ttataaatgt agagacaaac
     2401 cggtgtgtgc ggattttagg caaacaagaa aatattagag tgatgcaatt ggctttgttc
     2461 caggggatag ccaaaaagca tcgtgctgca actactatag aaatgaaagc ttctgaaaat
     2521 cctgttcttc agaatattca agctgaccca acaatagtct gtacatcatt caaaaagaat
     2581 agattttata tgtttaccaa acgagaacca gaagatacga aaagtgcaga ttctgatcga
     2641 gatgttttta atgagaaacc ttctaaagaa gaagtcatgg cagctactca agctgaagga
     2701 cctaaacgag tttcggacag tgccattatc cacaccagca tgggagacat tcacaccaaa
     2761 ctttttcctg ttgagtgccc taagacagtg gaaaacttct gtgttcacag cagaaatggt
     2821 tattataatg ggcatacatt tcaccgtata attaagggct ttatgattca gactggagat
     2881 ccaacaggta ctggtatggg aggagaaagc atatggggag gagaatttga agatgaattt
     2941 cattcaacat tacgacatga caggccgtac acactcagca tggctaacgc gggatcaaat
     3001 actaatggat cccagttttt cataacggta gtaccaacgc cttggcttga taataagcat
     3061 acagtatttg gacgagtgac taaaggaatg gaagttgtac agaggatctc caacgtcaaa
     3121 gtcaatccca aaacagataa gccctatgag gatgtcagca tcataaatat tactgtcaag
     3181 ggctcgagcg gtgggagctc cggtgtgagc ggctggcggc tgttcaagaa gattagctaa
     3241 gtttaaacgg ccgcgactct agactgcagg catgcaagct gatttggctg ctgccaccgc
     3301 tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     3361 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3421 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3481 caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt
     3541 aaaatcttaa ttaagacttc taatataact tcgtatagca tacattatac gaagttatgg
     3601 ttccactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     3661 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     3721 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3781 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3841 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3901 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3961 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4021 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4081 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4141 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4201 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4261 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4321 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4381 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4441 agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag
     4501 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4561 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4621 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4681 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4741 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4801 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4861 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4921 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4981 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5041 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     5101 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5161 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5221 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5281 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     5341 gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat
     5401 tgcggccatc gtgatggcta gagtttagcc ctcccacaca taaccagagg gcagcaattc
     5461 acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt tgatcccagg
     5521 atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc
     5581 atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag cagtgcccag
     5641 caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac cagtgaagat
     5701 gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga tggggatgct
     5761 gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca aaggcttggc
     5821 catggtggcg ctagtgtcag aagaatcgag ctttttgcaa aagcctaggc ctccaaaaaa
     5881 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     5941 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     6001 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     6061 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
     6121 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     6181 cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc ctctttcaga
     6241 ggttatttca ggccatggtg ctgcgc
//
