LOCUS       sequence-423337-        8199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR91. C-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8199)
  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 8199)
  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_WDR91:1-747
FEATURES             Location/Qualifiers
     source          1..8199
                     /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"
     regulatory      1079..1088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3374..3886
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3962..3980)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4017..4138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4028..4047)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4082..4101
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4383..4740
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4383..4574
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4406..4426)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4591..4726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4771..5808
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5787..5795
                     /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    5832..5880
                     /note="synthetic polyadenylation signal"
     promoter        6036..6140
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6141..6209,6210..7001)
                     /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(6359..6378)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7159..7747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7239..7258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7863..8146
                     /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 caccatggcg gaggccgtgg agcgcactga cgagctggtc cgggagtacc tgctcttccg
     1141 cgggttcacg cacacactgc ggcagctgga cgccgagatc aaggcggaca aggagaaggg
     1201 gttccgggtg gataagattg tggaccagct gcagcagtta atgcaggtgt atgacttggc
     1261 tgcccttcgg gattattgga gctacttgga gcgtcggctc ttcagccgct tggaggatat
     1321 atacagaccc acaatccaca agctgaaaac cagcctgttt cgattttatc ttgtctacac
     1381 aatccagaca aacagaaatg acaaggctca ggagttcttt gcaaagcagg ccacggaact
     1441 ccagaaccag gctgagtgga aggattggtt tgtcctgccc ttcctgccat ccccggacac
     1501 caaccccacc tttgctacct acttttctcg acagtgggct gacaccttca ttgtgtccct
     1561 gcacaacttc ctgagcgtcc tgtttcagtg catgccagtc cctgtgatcc tgaactttga
     1621 tgcggagtgt cagaggacta accaggttca agaagaaaat gaagttctgc gtcagaagct
     1681 ttttgcattg caagctgaaa tccaccgact gaagaaagag gagcaacagc cagaagagga
     1741 agaggccttg gtccaacaca aattgcctcc ttatgtctcc aacatggacc gcctggggga
     1801 ctcggaactt gccatggtgt gcagccaaag gaatgcctcc ctctcccagt cacctcgtgt
     1861 gggcttcctg tcctcgctgc tgcctcagag taagaagagc ccctcaaggt tgtcgcctgc
     1921 tcagggccct cctcaacctc agagctcggc caagaaagag tccttcggtg gtcagggcac
     1981 caagggaaag gacccgacgt ccggagccaa ggatgggaag agcctcctca gcgggctggc
     2041 cactggggag tccggttggt cacagcaccg gcagcggcgc ctgcaggacc atggcaagga
     2101 gaggaaggag cttttctcca caaccacttc ccagtgtgca gagaagaaac cagaagccag
     2161 tggcccagag gctgagccct gcccagagct ccacacggag ccagtggagc cactgactcg
     2221 ggcatcctcg gcaggccctg agggtggagg agtccgcccc gagcagccct ttattgtgct
     2281 gggacaggag gagtacgggg aacaccactc atccatcatg cactgcagag tggactgctc
     2341 tgggaggaga gtcgccagct tagacgtaga tggggtcatc aaagtgtggt ccttcaaccc
     2401 catcatgcag accaaagcat cctccatttc caaatcaccg ctgctgtctt tggaatgggc
     2461 caccaaacgg gacagactgc tcttgctggg cagtggtgtg ggaacagtgc gtctctatga
     2521 cacggaagcc aagaagaatc tctgtgaaat caatatcaac gacaacatgc ccagaatcct
     2581 gtctcttgcg tgcagcccca acggggcctc tttcgtctgt tcggcagcag ctccgagcct
     2641 cacttcccag gtggacttct cagcaccaga catcggcagc aagggcatga accaggttcc
     2701 tggcaggctg ctgctgtggg acacgaaaac catgaagcag cagctccagt tctccctgga
     2761 tccagaaccc attgctatca actgtacagc cttcaatcac aacgggaacc tgctggtcac
     2821 aggggcagct gatggcgtca tccggctgtt tgacatgcag cagcatgagt gcgcgatgag
     2881 ctggagggcc cactacgggg aggtctactc tgtggagttc agctatgatg agaacaccgt
     2941 gtacagcatc ggcgaggacg ggaagttcat ccagtggaac atccacaaga gtggcctcaa
     3001 ggtatccgag tacagcctcc cctcagatgc cacgggcccc tttgtgctgt ctggatacag
     3061 cggctacaag caggttcaag tccccagggg ccgactcttc gcttttgact cggagggaaa
     3121 ttacatgctg acatgttctg ccacaggcgg cgtcatctac aagctgggtg gcgatgagaa
     3181 ggttctggag agctgcttga gcctaggtgg ccaccgagcc cctgtggtca ccgtggactg
     3241 gagcactgcc atggactgtg ggacctgcct caccgcctcc atggatggca agatcaagct
     3301 gaccaccctc ctggcccata aagcctctag atttgggccc aattcctgca ggcgagctct
     3361 cggctcgagc ggcgtcttca cactcgaaga tttcgttggg gactggcgac agacagccgg
     3421 ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg
     3481 ggtgtccgta actccgatcc aaaggattgt cctgagcggt gaaaatgggc tgaagatcga
     3541 catccatgtc atcatcccgt atgaaggtct gagcggcgac caaatgggcc agatcgaaaa
     3601 aatttttaag gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgcactatgg
     3661 cacactggta atcgacgggg ttacgccgaa catgatcgac tatttcggac ggccgtatga
     3721 aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa
     3781 caaaattatc gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat
     3841 caacggagtg accggctggc ggctgtgcga acgcattctg gcgtaaggcc gcgactctag
     3901 ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     3961 accgctgagc aataactagc ataacccctt ggggcggccg cttcgagcag acatgataag
     4021 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4081 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4141 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt
     4201 aagcaagtaa aacctctaca aatgtggtaa aatcgaattt taacaaaata ttaacgctta
     4261 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     4321 cggatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc
     4381 ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4441 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4501 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4561 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4621 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4681 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4741 aagctcgatt cttctgacac tagcgccacc atgaagaagc ccgaactcac cgctaccagc
     4801 gttgaaaaat ttctcatcga gaagttcgac agtgtgagcg acctgatgca gttgtcggag
     4861 ggcgaagaga gccgagcctt cagcttcgat gtcggcggac gcggctatgt actgcgggtg
     4921 aatagctgcg ctgatggctt ctacaaagac cgctacgtgt accgccactt cgccagcgct
     4981 gcactaccca tccccgaagt gttggacatc ggcgagttca gcgagagcct gacatactgc
     5041 atcagtagac gcgcccaagg cgttactctc caagacctcc ccgaaacaga gctgcctgct
     5101 gtgttacagc ctgtcgccga agctatggat gctattgccg ccgccgacct cagtcaaacc
     5161 agcggcttcg gcccattcgg gccccaaggc atcggccagt acacaacctg gcgggatttc
     5221 atttgcgcca ttgctgatcc ccatgtctac cactggcaga ccgtgatgga cgacaccgtg
     5281 tccgccagcg tagctcaagc cctggacgaa ctgatgctgt gggccgaaga ctgtcccgag
     5341 gtgcgccacc tcgtccatgc cgacttcggc agcaacaacg tcctgaccga caacggccgc
     5401 atcaccgccg taatcgactg gtccgaagct atgttcgggg acagtcagta cgaggtggcc
     5461 aacatcttct tctggcggcc ctggctggct tgcatggagc agcagactcg ctacttcgag
     5521 cgccggcatc ccgagctggc cggcagccct cgtctgcgag cctacatgct gcgcatcggc
     5581 ctggatcagc tctaccagag cctcgtggac ggcaacttcg acgatgctgc ctgggctcaa
     5641 ggccgctgcg atgccatcgt ccgcagcggg gccggcaccg tcggtcgcac acaaatcgct
     5701 cgccggagcg cagccgtatg gaccgacggc tgcgtcgagg tgctggccga cagcggcaac
     5761 cgccggccca gtacacgacc gcgcgctaag gaggtaggtc gagtttagac tctagccatc
     5821 acgatggccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg
     5881 aatcgatagc gataaggatc ctctttgcgc ttgcgttttc ccttgtccag atagcccagt
     5941 agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac gtaatggttt
     6001 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6061 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6121 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6181 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6241 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6301 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttc aaagttctgc
     6361 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6421 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6481 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6541 acttacttct gacaactatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6601 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6661 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6721 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6781 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6841 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6901 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6961 agatcgctga gataggtgcc tcactgatta agcattggta attcgaaatg accgaccaag
     7021 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7081 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7141 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7201 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7261 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7321 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7381 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7441 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7501 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7561 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7621 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7681 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7741 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7801 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     7861 gtccggaaat acaggaacgc acgctggatg gcccttcgct gggatggtga aaccatgaaa
     7921 aatggcagct tcagtggatt aagtgggggt aatgtggcct gtaccctctg gttgcatagg
     7981 tattcatacg gttaaaattt atcaggcgcg attgcggcag tttttcgggt ggtttgttgc
     8041 catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta
     8101 agggattatg gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta
     8161 ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
