LOCUS       sequence-425999-        6222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PPWD1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6222)
  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 6222)
  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-N_PPWD1:1-646
FEATURES             Location/Qualifiers
     source          1..6222
                     /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"
     polyA_signal    3285..3406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3296..3315)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3521..3554)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3564..3611
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3643..4437
                     /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(3697..3716)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4307..4326
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4595..5183)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4675..4694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5309..5357
                     /note="synthetic polyadenylation signal"
     CDS             complement(5381..5779)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5776..5785
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5810..6167)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5824..5959
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5976..6167
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6124..6144
                     /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 ataggggtta gcgatcgcca tggtgagcgg
     1201 ctggcggctg ttcaagaaga ttagcgggag ctccggtggc tcgagcggta tggcggcgga
     1261 aagtggtagc gattttcagc agagacgtag aaggcgccgg gacccggagg aaccggaaaa
     1321 aacagaactc agcgaaagag agctggcagt agcagtggcg gtgtcccagg agaacgatga
     1381 ggagaacgaa gagcgctggg ttggaccttt acctgtggag gcaacactgg ccaagaagag
     1441 gaaagtctta gagtttgaaa gagtctatct tgataatctc cccagtgcat ccatgtatga
     1501 gcgcagttac atgcatagag atgttatcac ccatgtggta tgcaccaaaa cagattttat
     1561 tattactgcc agtcatgatg gacatgtcaa gttctggaaa aaaatagaag agggaattga
     1621 atttgttaaa cattttcgta gtcacctggg agttattgag agtattgcag ttagctctga
     1681 gggagcattg ttctgttctg tgggtgatga taaagcaatg aaggtgtttg atgtagtgaa
     1741 ctttgacatg atcaacatgc tgaaacttgg ctattttcct ggacagtgtg agtggatcta
     1801 ttgcccaggg gatgcaattt cttcagttgc tgcttccgaa aagagtacag gaaaaatttt
     1861 catttatgat ggccgaggag ataaccagcc acttcatatt tttgacaaac tccatacatc
     1921 acctcttact cagatacggc taaacccagt ttacaaagca gtagtgtctt ctgacaaatc
     1981 tgggatgatt gaatactgga ctgggcctcc tcatgaatat aaattcccca aaaatgtgaa
     2041 ctgggaatat aaaactgaca ctgatttata tgaatttgcc aagtgtaagg cttatccaac
     2101 cagcgtatgt ttttcaccag atgggaagaa aatagctact attggttctg atagaaaagt
     2161 tagaattttc agatttgtaa ctggaaaact catgagagtc tttgatgaat cactaagcat
     2221 gtttactgaa ctgcaacaga tgaggcaaca gttaccagac atggaatttg gccgacgaat
     2281 ggctgtagaa cgtgagttgg agaaggttga tgctgtaaga ttaattaata tagtttttga
     2341 tgaaactgga cacttcgtgc tgtatggaac aatgctgggc attaaagtta taaatgtaga
     2401 gacaaaccgg tgtgtgcgga ttttaggcaa acaagaaaat attagagtga tgcaattggc
     2461 tttgttccag gggatagcca aaaagcatcg tgctgcaact actatagaaa tgaaagcttc
     2521 tgaaaatcct gttcttcaga atattcaagc tgacccaaca atagtctgta catcattcaa
     2581 aaagaataga ttttatatgt ttaccaaacg agaaccagaa gatacgaaaa gtgcagattc
     2641 tgatcgagat gtttttaatg agaaaccttc taaagaagaa gtcatggcag ctactcaagc
     2701 tgaaggacct aaacgagttt cggacagtgc cattatccac accagcatgg gagacattca
     2761 caccaaactt tttcctgttg agtgccctaa gacagtggaa aacttctgtg ttcacagcag
     2821 aaatggttat tataatgggc atacatttca ccgtataatt aagggcttta tgattcagac
     2881 tggagatcca acaggtactg gtatgggagg agaaagcata tggggaggag aatttgaaga
     2941 tgaatttcat tcaacattac gacatgacag gccgtacaca ctcagcatgg ctaacgcggg
     3001 atcaaatact aatggatccc agtttttcat aacggtagta ccaacgcctt ggcttgataa
     3061 taagcataca gtatttggac gagtgactaa aggaatggaa gttgtacaga ggatctccaa
     3121 cgtcaaagtc aatcccaaaa cagataagcc ctatgaggat gtcagcatca taaatattac
     3181 tgtcaagtaa tctagaggtc aagcttcggg ggtttaaacc tgcaggcatg caagctgatt
     3241 tggctgctgc caccgctgag caaggccgct tcgagcagac atgataagat acattgatga
     3301 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3361 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3421 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     3481 cctctacaaa tgtggtaaaa tcttaattaa gacttctaat ataacttcgt atagcataca
     3541 ttatacgaag ttatggttcc actgttgaca attaatcatc ggcatagtat atcggcatag
     3601 tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga
     3661 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3721 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3781 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     3841 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3901 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3961 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4021 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4081 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4141 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4201 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4261 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4321 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4381 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc
     4441 gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt
     4501 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4561 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4621 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4681 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4741 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4801 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4861 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4921 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4981 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5041 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5101 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5161 cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5221 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5281 cctagatcct tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa
     5341 aataaagata ttttattgcg gccatcgtga tggctagagt ttagccctcc cacacataac
     5401 cagagggcag caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta
     5461 tggctttgat cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca
     5521 agatgcccct gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag
     5581 cagcagcagt gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat
     5641 tgacaccagt gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt
     5701 cagagatggg gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt
     5761 gagacaaagg cttggccatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc
     5821 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     5881 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     5941 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     6001 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     6061 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     6121 cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc
     6181 aagttcctct ttcagaggtt atttcaggcc atggtgctgc gc
//
