LOCUS       sequence-423340-        8568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human WDR81 fragment M728 to S1614. 
            N-terminal NanoLuc luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8568)
  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 8568)
  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_WDR81:728-1614
FEATURES             Location/Qualifiers
     source          1..8568
                     /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             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(4336..4354)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4386..4507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4397..4416)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4451..4470
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4752..5109
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4752..4943
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4775..4795)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4960..5095
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5140..6177
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             6156..6164
                     /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    6201..6249
                     /note="synthetic polyadenylation signal"
     promoter        6405..6509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6510..6578,6579..7370)
                     /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(6728..6747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7528..8116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7608..7627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8232..8515
                     /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 ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgcttc cgaattcatg caggccttgg
     1621 aggagctgga aaaaacagga aacttccttg ctaagggcct tggcggcctg ctggaggtcc
     1681 cagagcagcc tagggtacag cccgcggtac ccttgcagtg cctgttgcac cgagacatgc
     1741 aggccttggg tgtcctcctc gcggaaatgg tgtttgcgac gcgagtccga acgctccaac
     1801 ctgacgctcc tttgtgggtg cgctttcagg cagtgcgcgg actttgtaca cggcacccga
     1861 aggaggtgcc agtttccctt cagcctgtat tggatacatt gttgcagatg tccggaccgg
     1921 aggttcccat gggtgccgag cggggtaagc ttgaccaatt gttcgagtat cgaccggtga
     1981 gtcaaggact tccaccccca tgcccatcac agctgctgtc cccctttagc tcagtagttc
     2041 ccttccctcc ttatttcccg gcactgcata ggtttattct gctttatcag gctcgacgcg
     2101 tcgaggatga ggcccaaggc cgggagttgg tattcgccct ctggcaacag ctgggcgctg
     2161 tgcttaaaga cattacccca gagggacttg agatattgct gccctttgtc ctcagcctta
     2221 tgtccgagga acataccgcg gtgtatacag cctggtacct ctttgagcct gttgcgaagg
     2281 ctctgggacc taagaacgca aataagtacc tcctgaaacc cctcatagga gcttatgagt
     2341 ccccttgcca attgcacggg aggttttatt tgtatacaga ttgttttgtg gcccagctca
     2401 tggtaagatt gggcctgcag gcctttttga cgcacctgtt gccgcacgta ctccaagtat
     2461 tggccggagc tgaagcatca caggaagagt caaaggatct ggcgggagct gctgaagaag
     2521 aagagtccgg tcttccaggg gcgggcccag gcagttgcgc atttggagag gaaatcccga
     2581 tggatggcga acctccagcg tcttcaggtc tcggactccc agactataca agcggggtta
     2641 gcttccacga tcaggctgac ctccccgaga ccgaagattt ccaagctggt ctgtatgtaa
     2701 ccgaaagtcc acaaccgcaa gaagcggagg cagtgtcctt gggtcgactc agcgacaaat
     2761 cttctacatc cgaaacttct ctgggcgagg agagggcacc ggatgaagga ggggcccctg
     2821 ttgacaaaag ctcacttagg tccggagata gtagtcaaga tctcaaacag tctgaaggat
     2881 ccgaagaaga agaggaggaa gaggattcct gtgttgtgct ggaggaggaa gagggagaac
     2941 aggaggaagt taccggcgca agtgaactga cacttagtga tacggtgctc tcaatggaga
     3001 ccgtagtcgc cggtggctca ggaggagacg gggaagagga agaggaggcc ctgccagaac
     3061 aatcagaagg taaagaacaa aagatcctcc ttgacaccgc ctgtaagatg gtccgctggc
     3121 tctccgctaa gttgggacca acggttgcga gtcggcacgt ggcgcggaat ctcctgagac
     3181 tgctcacaag ctgctacgtt ggccctacta ggcaacagtt cacagtctcc tctggagaat
     3241 cccccccact ctccgctgga aacatctacc agaagagacc agtactgggt gacatcgtct
     3301 ctgggccagt gttgagctgc ttgttgcaca tcgcacgatt gtatggcgaa ccggtactta
     3361 cgtatcaata cctgccatat atttcttacc tcgtggcccc cggctcagct tctgggccct
     3421 cccgcctcaa tagccgaaaa gaagcgggtc tgcttgctgc tgtcacgttg actcaaaaaa
     3481 ttattgtata ccttagcgac acgacactga tggacatctt gccaagaatc agccatgagg
     3541 tgctgcttcc agtcttgtca tttttgacca gtcttgttac ggggtttccg tccggtgccc
     3601 aagcaaggac tatcctttgt gtaaaaacaa tatcattgat agcactcatt tgccttagga
     3661 ttggacagga gatggtacaa caacatttgt ccgaacccgt agctactttt ttccaagtct
     3721 ttagtcagct gcacgagctc cgccagcagg acttgaagtt ggatcctgct ggtaggggag
     3781 agggccaact gccccaggtc gttttcagcg atgggcaaca aaggccagtc gacccggccc
     3841 tcctggatga actccaaaaa gttttcaccc tggaaatggc ctacacgata tacgtgcctt
     3901 tttcatgcct tttgggtgat ataatcagaa aaattatccc taatcatgaa ctcgttggag
     3961 aacttgctgc cctgtatctt gaaagcattt cacccagttc aaggaatcca gcaagtgttg
     4021 aaccgacaat gcctggtacg ggacccgagt gggatcctca tggtggtggc tgcccgcaag
     4081 acgacgggca ttctggtaca ttcggtagcg tccttgtcgg caatcgcatt cagataccta
     4141 atgactcccg ccccgagaat cctggtcctt tgggaccgat atcaggggtg ggtgggggag
     4201 gactcggaag cggctccgac gacaatgccc tcaaacagga actgccacga agtgtccacg
     4261 gcctgtcata ataatctaga tttgggccca attcctgcag gattttgcgg ccgcttgctg
     4321 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggccgc ttcgagcaga
     4381 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4441 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4501 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     4561 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat
     4621 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4681 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     4741 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     4801 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4861 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4921 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4981 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5041 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5101 ttttgcaaaa agctcgattc ttctgacact agcgccacca tgaagaagcc cgaactcacc
     5161 gctaccagcg ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga cctgatgcag
     5221 ttgtcggagg gcgaagagag ccgagccttc agcttcgatg tcggcggacg cggctatgta
     5281 ctgcgggtga atagctgcgc tgatggcttc tacaaagacc gctacgtgta ccgccacttc
     5341 gccagcgctg cactacccat ccccgaagtg ttggacatcg gcgagttcag cgagagcctg
     5401 acatactgca tcagtagacg cgcccaaggc gttactctcc aagacctccc cgaaacagag
     5461 ctgcctgctg tgttacagcc tgtcgccgaa gctatggatg ctattgccgc cgccgacctc
     5521 agtcaaacca gcggcttcgg cccattcggg ccccaaggca tcggccagta cacaacctgg
     5581 cgggatttca tttgcgccat tgctgatccc catgtctacc actggcagac cgtgatggac
     5641 gacaccgtgt ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg ggccgaagac
     5701 tgtcccgagg tgcgccacct cgtccatgcc gacttcggca gcaacaacgt cctgaccgac
     5761 aacggccgca tcaccgccgt aatcgactgg tccgaagcta tgttcgggga cagtcagtac
     5821 gaggtggcca acatcttctt ctggcggccc tggctggctt gcatggagca gcagactcgc
     5881 tacttcgagc gccggcatcc cgagctggcc ggcagccctc gtctgcgagc ctacatgctg
     5941 cgcatcggcc tggatcagct ctaccagagc ctcgtggacg gcaacttcga cgatgctgcc
     6001 tgggctcaag gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt cggtcgcaca
     6061 caaatcgctc gccggagcgc agccgtatgg accgacggct gcgtcgaggt gctggccgac
     6121 agcggcaacc gccggcccag tacacgaccg cgcgctaagg aggtaggtcg agtttagact
     6181 ctagccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     6241 ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga
     6301 tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg
     6361 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6421 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6481 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6541 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6601 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6661 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttca
     6721 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6781 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6841 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6901 ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc gcttttttgc
     6961 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     7021 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     7081 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     7141 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7201 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7261 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7321 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ttcgaaatga
     7381 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7441 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7501 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7561 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7621 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7681 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7741 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7801 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7861 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7921 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7981 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     8041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     8101 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8161 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8221 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     8281 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     8341 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     8401 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     8461 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     8521 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
