LOCUS       sequence-423341-        8142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human WDR81 fragment M1 to L745. N-terminal 
            NanoLuc luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8142)
  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 8142)
  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_WDR81:1-745
FEATURES             Location/Qualifiers
     source          1..8142
                     /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(3910..3928)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3960..4081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3971..3990)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4025..4044
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4326..4683
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4326..4517
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4349..4369)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4534..4669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4714..5751
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5730..5738
                     /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    5775..5823
                     /note="synthetic polyadenylation signal"
     promoter        5979..6083
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6084..6152,6153..6944)
                     /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(6302..6321)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7102..7690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7182..7201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7806..8089
                     /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 gcacagggct
     1621 ctgggggtcg agaaggtgca ctccggacgc ctgctggtgg atggcacagc ccgccttccc
     1681 ctgacatgca ggaattgctt cgctccgtag aaagggatct ctctatagat ccacgccagc
     1741 ttgcgccggc acctggaggt acgcatgtcg tagccctggt accggccagg tggctcgcca
     1801 gtctgagaga tagacgactc ccattgggac cttgccccag ggctgaggga cttggtgagg
     1861 cggaggtgcg aaccttgctg cagaggtctg tccagaggct tcctgcgggt tggacaaggg
     1921 tagaggtgca cggtctccga aaacggcggt tgtcctatcc gctgggtggt ggcctccctt
     1981 ttgaggacgg aagttgcggc cctgaaactc tcacccggtt tatgcaagaa gtagccgcac
     2041 aaaactaccg aaatctgtgg aggcatgctt atcatacgta cgggcaaccg tattcccatt
     2101 cccctgcgcc gagcgcagta cccgcactgg atagtgtgcg acaagctctt cagcgggttt
     2161 atggttgttc ttttttgccc gttggcgaaa cgacgcaatg cccctcttac gctcgagagg
     2221 gaccttgtcc tccacgaggg agccctgcct gtccttcctt gttgcgggca gaggctctcc
     2281 tcgagagtcc ggagatgctg tatgttgtgc acccttacgt gcagtttagc ctgcatgatg
     2341 tggtgacctt cagcccagca aagttgacta atagtcaggc aaaagtgctc tttatcttgt
     2401 ttcgagtcct gcgagccatg gacgcttgcc accggcaagg tctcgcctgc ggtgcattgt
     2461 ctctttacca tattgccgtc gatgagaagc tctgcagtga gcttagattg gatctgagtg
     2521 cttacgaacg gcctgaggag gacgagaatg aggaggcccc agtggcgcga gacgaagcgg
     2581 gtatcgtatc ccaagaggag cagggtggcc aacccggtca acccacgggt caagaagagc
     2641 ttagatctct cgtgcttgac tgggtccacg gcagaatctc caacttccac tatttgatgc
     2701 aactcaacag actggcgggg cggaggcagg gtgacccgaa ttaccatcca gtcctgccgt
     2761 gggtggtaga tttcacaacc ccacatggtc ggttccgaga cctccggaaa tctaagttca
     2821 ggttgaacaa gggcgataag caactggact ttacctatga aatgactcgg caggcctttg
     2881 tagcgggcgg agcagggggg ggagagcctc cccacgtgcc gcatcatatc tcagatgtcc
     2941 tgtccgacat cacctactat gtgtacaaag ccagaaggac tcctcggtct gttttgtgtg
     3001 gacacgttcg cgcccaatgg gaaccccatg aatatccagc gtcaatggag cgaatgcaga
     3061 attggacgcc agacgaatgt attcctgagt tctacaccga cccgagtata tttcgatcta
     3121 ttcaccccga tatgcctgac ttggatgtgc ccgcatggtg ctcaagtagc caggagttcg
     3181 tagctgccca tcgcgccttg ttggaatcac gggaagtaag ccgggatctg caccattgga
     3241 ttgaccttac attcggatac aaattgcagg gtaaagaggc agttaaggaa aagaacgttt
     3301 gtcttcatct ggttgacgca cacacgcatc ttgcgtccta cggggttgtc caattgttcg
     3361 atcaacccca tccgcaaagg ctcgctggcg ccccagctct tgccccagaa ccgccgctta
     3421 tacctaaact tctggtgcag acgatacaag aaaccactgg tcgagaggat tttaccgaaa
     3481 atcctgggca attgcccaac ggagtcggca gaccagtatt ggaagcaacc ccctgtgaag
     3541 cttcttggac tagggataga cctgtggccg gtgaagatga cctggagcaa gccaccgaag
     3601 cactggatag tataagcctc gcgggaaagg ccggggacca gctcggaagc agctctcaag
     3661 ccagtcctgg cctcctcagc ttttcagtcg cgagtgcctc aagacctggg cggcgcaaca
     3721 aagccgccgg tgccgaccca ggcgaagggg aggaagggag gatattgctt ccagaagggt
     3781 tcaatcctat gcaggccttg gaggagctgg aaaaaacagg aaacttcctt gctaagggcc
     3841 tttaataatc tagatttggg cccaattcct gcaggatttt gcggccgctt gctgagttgg
     3901 ctgctgccac cgctgagcaa taactagcat aaccccttgg ccgcttcgag cagacatgat
     3961 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4021 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4081 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     4141 tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc
     4201 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4261 acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
     4321 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4381 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4441 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4501 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4561 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4621 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4681 aaaaagctcg attcttctga cactagcgcc accatgaaga agcccgaact caccgctacc
     4741 agcgttgaaa aatttctcat cgagaagttc gacagtgtga gcgacctgat gcagttgtcg
     4801 gagggcgaag agagccgagc cttcagcttc gatgtcggcg gacgcggcta tgtactgcgg
     4861 gtgaatagct gcgctgatgg cttctacaaa gaccgctacg tgtaccgcca cttcgccagc
     4921 gctgcactac ccatccccga agtgttggac atcggcgagt tcagcgagag cctgacatac
     4981 tgcatcagta gacgcgccca aggcgttact ctccaagacc tccccgaaac agagctgcct
     5041 gctgtgttac agcctgtcgc cgaagctatg gatgctattg ccgccgccga cctcagtcaa
     5101 accagcggct tcggcccatt cgggccccaa ggcatcggcc agtacacaac ctggcgggat
     5161 ttcatttgcg ccattgctga tccccatgtc taccactggc agaccgtgat ggacgacacc
     5221 gtgtccgcca gcgtagctca agccctggac gaactgatgc tgtgggccga agactgtccc
     5281 gaggtgcgcc acctcgtcca tgccgacttc ggcagcaaca acgtcctgac cgacaacggc
     5341 cgcatcaccg ccgtaatcga ctggtccgaa gctatgttcg gggacagtca gtacgaggtg
     5401 gccaacatct tcttctggcg gccctggctg gcttgcatgg agcagcagac tcgctacttc
     5461 gagcgccggc atcccgagct ggccggcagc cctcgtctgc gagcctacat gctgcgcatc
     5521 ggcctggatc agctctacca gagcctcgtg gacggcaact tcgacgatgc tgcctgggct
     5581 caaggccgct gcgatgccat cgtccgcagc ggggccggca ccgtcggtcg cacacaaatc
     5641 gctcgccgga gcgcagccgt atggaccgac ggctgcgtcg aggtgctggc cgacagcggc
     5701 aaccgccggc ccagtacacg accgcgcgct aaggaggtag gtcgagttta gactctagcc
     5761 atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     5821 gtgaatcgat agcgataagg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc
     5881 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtaatgg
     5941 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     6001 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6061 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6121 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6181 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6241 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact ttcaaagttc
     6301 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6361 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6421 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6481 ccaacttact tctgacaact atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6541 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6601 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6661 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6721 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6781 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6841 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6901 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaattcgaa atgaccgacc
     6961 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7021 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7081 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7141 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7201 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7261 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7321 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7381 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7441 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7501 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7561 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7621 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7681 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7741 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7801 atagtccgga aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg
     7861 aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat
     7921 aggtattcat acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt
     7981 tgccattttt acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa
     8041 ttaagggatt atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca
     8101 gtactccggc cacgatgcgt ccggcgtaga ggatcgagat ct
//
