LOCUS       sequence-423315-        7461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PWP1. C-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7461)
  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 7461)
  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-C_PWP1:1-501
FEATURES             Location/Qualifiers
     source          1..7461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2636..3148
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3224..3242)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3279..3400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3290..3309)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3344..3363
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3645..4002
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3645..3836
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3668..3688)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3853..3988
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4033..5070
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5049..5057
                     /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    5094..5142
                     /note="synthetic polyadenylation signal"
     promoter        5298..5402
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5403..5471,5472..6263)
                     /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(5621..5640)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6421..7009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6501..6520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7125..7408
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcaaa gcgatcgctt ccgaattcgc
     1081 caccatgaac cgcagccgcc aggtgacgtg cgtggcctgg gtccgctgcg gcgtggccaa
     1141 agagacacca gacaaggtag agctgagtaa agaagaagta aaacgcctca ttgctgaggc
     1201 aaaggagaaa ttgcaagaag aaggtggtgg cagtgatgaa gaggagacag gcagtccttc
     1261 agaagatggc atgcagagtg cacgcaccca ggcacgccca agagagcccc tggaggatgg
     1321 tgacccagag gatgacagga cgcttgatga tgatgagctg gctgagtacg acttagataa
     1381 atatgatgag gaaggtgacc cagatgctga gactcttggt gaatctctct tgggtcttac
     1441 ggtctacggg agtaatgatc aagatcctta cgttactctg aaagatacag aacaatatga
     1501 acgtgaagat ttcttgatta agcccagtga taatcttata gtttgtggcc gagctgaaca
     1561 ggaccagtgc aatttagacg tgcatgttta taatcaagaa gaagactctt tttatgtaca
     1621 ccatgatata ctcttgtctg catatcctct gagtgtggaa tggctgaatt ttgatcctag
     1681 cccagatgat tctactggaa attacattgc tgtaggaaac atgacccctg ttattgaagt
     1741 gtgggacctt gatatagtgg actctttaga gccagtcttc acactcggaa gtaaactttc
     1801 aaaaaagaag aaaaagaaag gaaagaagag ttcctcagca gaagggcata ccgatgctgt
     1861 ccttgacctt tcatggaata agctaatcag aaatgtttta gcaagtgcat cagctgacaa
     1921 cactgtaatt ctgtgggata tgtccttggg gaaaccagca gctagcctcg ctgtacacac
     1981 agacaaggtc caaacactgc agtttcatcc atttgaagca cagactctga tttctggctc
     2041 atatgataag tcagtggctt tgtatgactg ccgaagtcca gatgaaagcc atcgaatgtg
     2101 gcgattcagt gggcagatag agagagtgac ttggaatcac ttttcacctt gtcatttctt
     2161 ggccagtaca gatgacggct ttgtatataa tttggatgca cgttcagata agccaatttt
     2221 tacacttaat gcacacaatg atgaaatctc tggtcttgat cttagcagtc aaatcaaggg
     2281 ctgtctcgtg actgcttcag ctgacaaata cgtgaagatc tgggacatct taggagatag
     2341 gccaagtcta gttcattcta gggacatgaa aatgggagtt ctcttctgtt cttcatgttg
     2401 ccctgatttg ccatttattt atgcccttcg aggtcaaaaa gaagggcttc gggtctggga
     2461 tataagcaca gtctcttcag taaatgaagc atttggaaga cgagagaggc ttgttcttgg
     2521 gagtgcaaga aattcatcta ttagtggccc ttttggcagc aggagctcag atacacccat
     2581 ggagtcttct agatttgggc ccaattcctg caggcgagct ctcggctcga gcggcgtctt
     2641 cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt
     2701 ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat
     2761 ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc
     2821 gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta
     2881 ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg
     2941 ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga
     3001 cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg
     3061 cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg
     3121 gcggctgtgc gaacgcattc tggcgtaagg ccgcgactct agggcatgca agctgatccg
     3181 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     3241 gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3301 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3361 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3421 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     3481 caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     3541 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca
     3601 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     3661 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3721 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3781 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3841 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3901 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3961 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcga ttcttctgac
     4021 actagcgcca ccatgaagaa gcccgaactc accgctacca gcgttgaaaa atttctcatc
     4081 gagaagttcg acagtgtgag cgacctgatg cagttgtcgg agggcgaaga gagccgagcc
     4141 ttcagcttcg atgtcggcgg acgcggctat gtactgcggg tgaatagctg cgctgatggc
     4201 ttctacaaag accgctacgt gtaccgccac ttcgccagcg ctgcactacc catccccgaa
     4261 gtgttggaca tcggcgagtt cagcgagagc ctgacatact gcatcagtag acgcgcccaa
     4321 ggcgttactc tccaagacct ccccgaaaca gagctgcctg ctgtgttaca gcctgtcgcc
     4381 gaagctatgg atgctattgc cgccgccgac ctcagtcaaa ccagcggctt cggcccattc
     4441 gggccccaag gcatcggcca gtacacaacc tggcgggatt tcatttgcgc cattgctgat
     4501 ccccatgtct accactggca gaccgtgatg gacgacaccg tgtccgccag cgtagctcaa
     4561 gccctggacg aactgatgct gtgggccgaa gactgtcccg aggtgcgcca cctcgtccat
     4621 gccgacttcg gcagcaacaa cgtcctgacc gacaacggcc gcatcaccgc cgtaatcgac
     4681 tggtccgaag ctatgttcgg ggacagtcag tacgaggtgg ccaacatctt cttctggcgg
     4741 ccctggctgg cttgcatgga gcagcagact cgctacttcg agcgccggca tcccgagctg
     4801 gccggcagcc ctcgtctgcg agcctacatg ctgcgcatcg gcctggatca gctctaccag
     4861 agcctcgtgg acggcaactt cgacgatgct gcctgggctc aaggccgctg cgatgccatc
     4921 gtccgcagcg gggccggcac cgtcggtcgc acacaaatcg ctcgccggag cgcagccgta
     4981 tggaccgacg gctgcgtcga ggtgctggcc gacagcggca accgccggcc cagtacacga
     5041 ccgcgcgcta aggaggtagg tcgagtttag actctagcca tcacgatggc cgcaataaaa
     5101 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga
     5161 tcctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     5221 gtcagcaccg tttctgcgga ctggctttct acgtaatggt ttcttagacg tcaggtggca
     5281 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5341 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5401 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5461 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5521 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5581 ccgaagaacg ttttccaatg atgagcactt tcaaagttct gctatgtggc gcggtattat
     5641 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5701 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5761 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacta
     5821 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5881 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5941 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6001 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6061 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6121 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6181 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6241 cctcactgat taagcattgg taattcgaaa tgaccgacca agcgacgccc aaccggtatc
     6301 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6361 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6421 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6481 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6541 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6601 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6661 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6721 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6781 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6841 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6901 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6961 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt
     7021 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7081 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac
     7141 gcacgctgga tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga
     7201 ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat
     7261 ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct
     7321 gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc
     7381 acttactgtc tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc
     7441 cggcgtagag gatcgagatc t
//
