LOCUS       sequence-425888-        5787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PWP1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5787)
  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 5787)
  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_PWP1:1-501
FEATURES             Location/Qualifiers
     source          1..5787
                     /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    2850..2971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2861..2880)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3086..3119)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3129..3176
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3208..4002
                     /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(3262..3281)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3872..3891
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4160..4748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4240..4259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4874..4922
                     /note="synthetic polyadenylation signal"
     CDS             complement(4946..5344)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5341..5350
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5375..5732)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5389..5524
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5541..5732
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5689..5709
                     /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 tgaaccgcag
     1261 ccgccaggtg acgtgcgtgg cctgggtccg ctgcggcgtg gccaaagaga caccagacaa
     1321 ggtagagctg agtaaagaag aagtaaaacg cctcattgct gaggcaaagg agaaattgca
     1381 agaagaaggt ggtggcagtg atgaagagga gacaggcagt ccttcagaag atggcatgca
     1441 gagtgcacgc acccaggcac gcccaagaga gcccctggag gatggtgacc cagaggatga
     1501 caggacgctt gatgatgatg agctggctga gtacgactta gataaatatg atgaggaagg
     1561 tgacccagat gctgagactc ttggtgaatc tctcttgggt cttacggtct acgggagtaa
     1621 tgatcaagat ccttacgtta ctctgaaaga tacagaacaa tatgaacgtg aagatttctt
     1681 gattaagccc agtgataatc ttatagtttg tggccgagct gaacaggacc agtgcaattt
     1741 agacgtgcat gtttataatc aagaagaaga ctctttttat gtacaccatg atatactctt
     1801 gtctgcatat cctctgagtg tggaatggct gaattttgat cctagcccag atgattctac
     1861 tggaaattac attgctgtag gaaacatgac ccctgttatt gaagtgtggg accttgatat
     1921 agtggactct ttagagccag tcttcacact cggaagtaaa ctttcaaaaa agaagaaaaa
     1981 gaaaggaaag aagagttcct cagcagaagg gcataccgat gctgtccttg acctttcatg
     2041 gaataagcta atcagaaatg ttttagcaag tgcatcagct gacaacactg taattctgtg
     2101 ggatatgtcc ttggggaaac cagcagctag cctcgctgta cacacagaca aggtccaaac
     2161 actgcagttt catccatttg aagcacagac tctgatttct ggctcatatg ataagtcagt
     2221 ggctttgtat gactgccgaa gtccagatga aagccatcga atgtggcgat tcagtgggca
     2281 gatagagaga gtgacttgga atcacttttc accttgtcat ttcttggcca gtacagatga
     2341 cggctttgta tataatttgg atgcacgttc agataagcca atttttacac ttaatgcaca
     2401 caatgatgaa atctctggtc ttgatcttag cagtcaaatc aagggctgtc tcgtgactgc
     2461 ttcagctgac aaatacgtga agatctggga catcttagga gataggccaa gtctagttca
     2521 ttctagggac atgaaaatgg gagttctctt ctgttcttca tgttgccctg atttgccatt
     2581 tatttatgcc cttcgaggtc aaaaagaagg gcttcgggtc tgggatataa gcacagtctc
     2641 ttcagtaaat gaagcatttg gaagacgaga gaggcttgtt cttgggagtg caagaaattc
     2701 atctattagt ggcccttttg gcagcaggag ctcagataca cccatggagt cttaatctag
     2761 aggtcaagct tcgggggttt aaacctgcag gcatgcaagc tgatttggct gctgccaccg
     2821 ctgagcaagg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     2881 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2941 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3001 tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg
     3061 taaaatctta attaagactt ctaatataac ttcgtatagc atacattata cgaagttatg
     3121 gttccactgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     3181 tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca cgcaggttct
     3241 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3301 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3361 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3421 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3481 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3541 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3601 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3661 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3721 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3781 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3841 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     3901 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     3961 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gattcgaaat gaccgaccaa
     4021 gcgacgccca accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4081 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4141 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4201 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4261 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4321 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4381 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4441 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4501 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4561 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     4621 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4681 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4741 gatttcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4801 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttat
     4861 agtccggaat cgtcacacaa aaaaccaaca cacagatgta atgaaaataa agatatttta
     4921 ttgcggccat cgtgatggct agagtttagc cctcccacac ataaccagag ggcagcaatt
     4981 cacgaatccc aactgccgtc ggctgtccat cactgtcctt cactatggct ttgatcccag
     5041 gatgcagatc gagaagcacc tgtcggcacc gtccgcaggg gctcaagatg cccctgttct
     5101 catttccgat cgcgacgata caagtcaggt tgccagctgc cgcagcagca gcagtgccca
     5161 gcaccacgag ttctgcacaa ggtcccccag taaaatgata tacattgaca ccagtgaaga
     5221 tgcggccgtc gctagagaga gctgcgctgg cgacgctgta gtcttcagag atggggatgc
     5281 tgttgattgt agccgttgct ctttcaatga gggtggattc ttcttgagac aaaggcttgg
     5341 ccatggtggc gctagtgtca gaagaatcga gctttttgca aaagcctagg cctccaaaaa
     5401 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     5461 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     5521 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     5581 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     5641 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     5701 acacattcca cagctggttc tttccgcctc agaaggtacc taaccaagtt cctctttcag
     5761 aggttatttc aggccatggt gctgcgc
//
