LOCUS       sequence-426254-        7410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PWP1. N-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7410)
  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 7410)
  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: pNLF1-N_PWP1:1-501
FEATURES             Location/Qualifiers
     source          1..7410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          136..268
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     251..270
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     312..331
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        312..330
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             344..856
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(2457..2475)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2507..2628
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2518..2537)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2572..2591
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2873..3230
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2873..3064
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2896..2916)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3081..3216
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3261..4298
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4277..4285
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4322..4370
                     /note="synthetic polyadenylation signal"
     promoter        4526..4630
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4631..4699,4700..5491)
                     /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     complement(4849..4868)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5649..6237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5729..5748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6353..6636
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        6827..7206
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7207..7410
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7361..7381
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggtttagtga accgtcagat cactagaagc tttattgcgg tagtttatca cagttaaatt
       61 gctaacgcag tcagtgcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg
      121 tgaggcactg ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa
      181 ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact
      241 gacatccact ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa
      301 ggctagagta ttaatacgac tcactatagg gctagcgctc accatggtct tcacactcga
      361 agatttcgtt ggggactggc gacagacagc cggctacaac ctggaccaag tccttgaaca
      421 gggaggtgtg tccagtttgt ttcagaatct cggggtgtcc gtaactccga tccaaaggat
      481 tgtcctgagc ggtgaaaatg ggctgaagat cgacatccat gtcatcatcc cgtatgaagg
      541 tctgagcggc gaccaaatgg gccagatcga aaaaattttt aaggtggtgt accctgtgga
      601 tgatcatcac tttaaggtga tcctgcacta tggcacactg gtaatcgacg gggttacgcc
      661 gaacatgatc gactatttcg gacggccgta tgaaggcatc gccgtgttcg acggcaaaaa
      721 gatcactgta acagggaccc tgtggaacgg caacaaaatt atcgacgagc gcctgatcaa
      781 ccccgacggc tccctgctgt tccgagtaac catcaacgga gtgaccggct ggcggctgtg
      841 cgaacgcatt ctggcgggct cgagcggcgc gatcgcttcc gaattcatga accgcagccg
      901 ccaggtgacg tgcgtggcct gggtccgctg cggcgtggcc aaagagacac cagacaaggt
      961 agagctgagt aaagaagaag taaaacgcct cattgctgag gcaaaggaga aattgcaaga
     1021 agaaggtggt ggcagtgatg aagaggagac aggcagtcct tcagaagatg gcatgcagag
     1081 tgcacgcacc caggcacgcc caagagagcc cctggaggat ggtgacccag aggatgacag
     1141 gacgcttgat gatgatgagc tggctgagta cgacttagat aaatatgatg aggaaggtga
     1201 cccagatgct gagactcttg gtgaatctct cttgggtctt acggtctacg ggagtaatga
     1261 tcaagatcct tacgttactc tgaaagatac agaacaatat gaacgtgaag atttcttgat
     1321 taagcccagt gataatctta tagtttgtgg ccgagctgaa caggaccagt gcaatttaga
     1381 cgtgcatgtt tataatcaag aagaagactc tttttatgta caccatgata tactcttgtc
     1441 tgcatatcct ctgagtgtgg aatggctgaa ttttgatcct agcccagatg attctactgg
     1501 aaattacatt gctgtaggaa acatgacccc tgttattgaa gtgtgggacc ttgatatagt
     1561 ggactcttta gagccagtct tcacactcgg aagtaaactt tcaaaaaaga agaaaaagaa
     1621 aggaaagaag agttcctcag cagaagggca tactgatgct gtccttgacc tttcatggaa
     1681 taagctaatc agaaatgttt tagcaagtgc atcagctgac aacactgtaa ttctgtggga
     1741 tatgtccttg gggaaaccag cagctagcct cgctgtacac acagacaagg tccaaacact
     1801 gcagtttcat ccatttgaag cacagactct gatttctggc tcatatgata agtcagtggc
     1861 tttgtatgac tgccgaagtc cagatgaaag ccatcgaatg tggcgattca gtgggcagat
     1921 agagagagtg acttggaatc acttttcacc ttgtcatttc ttggccagta cagatgacgg
     1981 ctttgtatat aatttggatg cacgttcaga taagccaatt tttacactta atgcacacaa
     2041 tgatgaaatc tctggtcttg atcttagcag tcaaatcaag ggctgtctcg tgactgcttc
     2101 agctgacaaa tacgtgaaga tctgggacat cttaggagat aggccaagtc tagttcattc
     2161 tagggacatg aaaatgggag ttctcttctg ttcttcatgt tgccctgatt tgccatttat
     2221 ttatgccctt cgaggtcaaa aagaagggct tcgggtctgg gatataagca cagtctcttc
     2281 agtaaatgaa gcatttggaa gacgagagag gcttgttctt gggagtgcaa gaaattcatc
     2341 tattagtggc ccttttggca gcaggagctc agatacaccc atggagtctt aataatctag
     2401 atttgggccc aattcctgca ggattttgcg gccgcttgct gagttggctg ctgccaccgc
     2461 tgagcaataa ctagcataac cccttggccg cttcgagcag acatgataag atacattgat
     2521 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2581 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2641 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa
     2701 aacctctaca aatgtggtaa aatcgaattt taacaaaata ttaacgctta caatttcctg
     2761 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg
     2821 cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg
     2881 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3001 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3061 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3121 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3181 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcgatt
     3241 cttctgacac tagcgccacc atgaagaagc ccgaactcac cgctaccagc gttgaaaaat
     3301 ttctcatcga gaagttcgac agtgtgagcg acctgatgca gttgtcggag ggcgaagaga
     3361 gccgagcctt cagcttcgat gtcggcggac gcggctatgt actgcgggtg aatagctgcg
     3421 ctgatggctt ctacaaagac cgctacgtgt accgccactt cgccagcgct gcactaccca
     3481 tccccgaagt gttggacatc ggcgagttca gcgagagcct gacatactgc atcagtagac
     3541 gcgcccaagg cgttactctc caagacctcc ccgaaacaga gctgcctgct gtgttacagc
     3601 ctgtcgccga agctatggat gctattgccg ccgccgacct cagtcaaacc agcggcttcg
     3661 gcccattcgg gccccaaggc atcggccagt acacaacctg gcgggatttc atttgcgcca
     3721 ttgctgatcc ccatgtctac cactggcaga ccgtgatgga cgacaccgtg tccgccagcg
     3781 tagctcaagc cctggacgaa ctgatgctgt gggccgaaga ctgtcccgag gtgcgccacc
     3841 tcgtccatgc cgacttcggc agcaacaacg tcctgaccga caacggccgc atcaccgccg
     3901 taatcgactg gtccgaagct atgttcgggg acagtcagta cgaggtggcc aacatcttct
     3961 tctggcggcc ctggctggct tgcatggagc agcagactcg ctacttcgag cgccggcatc
     4021 ccgagctggc cggcagccct cgtctgcgag cctacatgct gcgcatcggc ctggatcagc
     4081 tctaccagag cctcgtggac ggcaacttcg acgatgctgc ctgggctcaa ggccgctgcg
     4141 atgccatcgt ccgcagcggg gccggcaccg tcggtcgcac acaaatcgct cgccggagcg
     4201 cagccgtatg gaccgacggc tgcgtcgagg tgctggccga cagcggcaac cgccggccca
     4261 gtacacgacc gcgcgctaag gaggtaggtc gagtttagac tctagccatc acgatggccg
     4321 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc
     4381 gataaggatc ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt
     4441 catccggggt cagcaccgtt tctgcggact ggctttctac gtaatggttt cttagacgtc
     4501 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4561 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4621 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4681 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4741 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4801 ttttcgcccc gaagaacgtt ttccaatgat gagcactttc aaagttctgc tatgtggcgc
     4861 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4921 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4981 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5041 gacaactatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5101 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5161 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5221 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5281 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5341 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5401 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5461 gataggtgcc tcactgatta agcattggta attcgaaatg accgaccaag cgacgcccaa
     5521 ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5581 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5641 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5701 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5761 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5821 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5881 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5941 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6001 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6061 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6121 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6181 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa
     6241 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6301 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata gtccggaaat
     6361 acaggaacgc acgctggatg gcccttcgct gggatggtga aaccatgaaa aatggcagct
     6421 tcagtggatt aagtgggggt aatgtggcct gtaccctctg gttgcatagg tattcatacg
     6481 gttaaaattt atcaggcgcg attgcggcag tttttcgggt ggtttgttgc catttttacc
     6541 tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta agggattatg
     6601 gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta ctccggccac
     6661 gatgcgtccg gcgtagagga tcgagatctt caatattggc cattagccat attattcatt
     6721 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     6781 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttggca ttgattattg
     6841 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     6901 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     6961 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     7021 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     7081 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     7141 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     7201 accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg
     7261 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     7321 cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt
     7381 gtacggtggg aggtctatat aagcagagct
//
