LOCUS       sequence-423339-        7119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human WDR81 fragment E1211 to S1614. 
            N-terminal NanoLuc luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7119)
  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 7119)
  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_WDR81:1211-1614
FEATURES             Location/Qualifiers
     source          1..7119
                     /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(2887..2905)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2937..3058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2948..2967)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3002..3021
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3303..3660
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3303..3494
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3326..3346)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3511..3646
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3691..4728
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4707..4715
                     /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    4752..4800
                     /note="synthetic polyadenylation signal"
     promoter        4956..5060
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5061..5129,5130..5921)
                     /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(5279..5298)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6079..6667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6159..6178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6783..7066
                     /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 cgaattcgaa caatcagaag
     1621 gtaaagaaca aaagatcctc cttgacaccg cctgtaagat ggtccgctgg ctctccgcta
     1681 agttgggacc aacggttgcg agtcggcacg tggcgcggaa tctcctgaga ctgctcacaa
     1741 gctgctacgt tggccctact aggcaacagt tcacagtctc ctctggagaa tcccccccac
     1801 tctccgctgg aaacatctac cagaagagac cagtactggg tgacatcgtc tctgggccag
     1861 tgttgagctg cttgttgcac atcgcacgat tgtatggcga accggtactt acgtatcaat
     1921 acctgccata tatttcttac ctcgtggccc ccggctcagc ttctgggccc tcccgcctca
     1981 atagccgaaa agaagcgggt ctgcttgctg ctgtcacgtt gactcaaaaa attattgtat
     2041 accttagcga cacgacactg atggacatct tgccaagaat cagccatgag gtgctgcttc
     2101 cagtcttgtc atttttgacc agtcttgtta cggggtttcc gtccggtgcc caagcaagga
     2161 ctatcctttg tgtaaaaaca atatcattga tagcactcat ttgccttagg attggacagg
     2221 agatggtaca acaacatttg tccgaacccg tagctacttt tttccaagtc tttagtcagc
     2281 tgcacgagct ccgccagcag gacttgaagt tggatcctgc tggtagggga gagggccaac
     2341 tgccccaggt cgttttcagc gatgggcaac aaaggccagt cgacccggcc ctcctggatg
     2401 aactccaaaa agttttcacc ctggaaatgg cctacacgat atacgtgcct ttttcatgcc
     2461 ttttgggtga tataatcaga aaaattatcc ctaatcatga actcgttgga gaacttgctg
     2521 ccctgtatct tgaaagcatt tcacccagtt caaggaatcc agcaagtgtt gaaccgacaa
     2581 tgcctggtac gggacccgag tgggatcctc atggtggtgg ctgcccgcaa gacgacgggc
     2641 attctggtac attcggtagc gtccttgtcg gcaatcgcat tcagatacct aatgactccc
     2701 gccccgagaa tcctggtcct ttgggaccga tatcaggggt gggtggggga ggactcggaa
     2761 gcggctccga cgacaatgcc ctcaaacagg aactgccacg aagtgtccac ggcctgtcat
     2821 aataatctag atttgggccc aattcctgca ggattttgcg gccgcttgct gagttggctg
     2881 ctgccaccgc tgagcaataa ctagcataac cccttggccg cttcgagcag acatgataag
     2941 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3001 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3061 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt
     3121 aagcaagtaa aacctctaca aatgtggtaa aatcgaattt taacaaaata ttaacgctta
     3181 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     3241 cggatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc
     3301 ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3361 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3421 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3481 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3541 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3601 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3661 aagctcgatt cttctgacac tagcgccacc atgaagaagc ccgaactcac cgctaccagc
     3721 gttgaaaaat ttctcatcga gaagttcgac agtgtgagcg acctgatgca gttgtcggag
     3781 ggcgaagaga gccgagcctt cagcttcgat gtcggcggac gcggctatgt actgcgggtg
     3841 aatagctgcg ctgatggctt ctacaaagac cgctacgtgt accgccactt cgccagcgct
     3901 gcactaccca tccccgaagt gttggacatc ggcgagttca gcgagagcct gacatactgc
     3961 atcagtagac gcgcccaagg cgttactctc caagacctcc ccgaaacaga gctgcctgct
     4021 gtgttacagc ctgtcgccga agctatggat gctattgccg ccgccgacct cagtcaaacc
     4081 agcggcttcg gcccattcgg gccccaaggc atcggccagt acacaacctg gcgggatttc
     4141 atttgcgcca ttgctgatcc ccatgtctac cactggcaga ccgtgatgga cgacaccgtg
     4201 tccgccagcg tagctcaagc cctggacgaa ctgatgctgt gggccgaaga ctgtcccgag
     4261 gtgcgccacc tcgtccatgc cgacttcggc agcaacaacg tcctgaccga caacggccgc
     4321 atcaccgccg taatcgactg gtccgaagct atgttcgggg acagtcagta cgaggtggcc
     4381 aacatcttct tctggcggcc ctggctggct tgcatggagc agcagactcg ctacttcgag
     4441 cgccggcatc ccgagctggc cggcagccct cgtctgcgag cctacatgct gcgcatcggc
     4501 ctggatcagc tctaccagag cctcgtggac ggcaacttcg acgatgctgc ctgggctcaa
     4561 ggccgctgcg atgccatcgt ccgcagcggg gccggcaccg tcggtcgcac acaaatcgct
     4621 cgccggagcg cagccgtatg gaccgacggc tgcgtcgagg tgctggccga cagcggcaac
     4681 cgccggccca gtacacgacc gcgcgctaag gaggtaggtc gagtttagac tctagccatc
     4741 acgatggccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg
     4801 aatcgatagc gataaggatc ctctttgcgc ttgcgttttc ccttgtccag atagcccagt
     4861 agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac gtaatggttt
     4921 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4981 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5041 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5101 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5161 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5221 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttc aaagttctgc
     5281 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5341 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5401 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5461 acttacttct gacaactatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5521 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5581 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5641 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5701 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5761 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5821 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5881 agatcgctga gataggtgcc tcactgatta agcattggta attcgaaatg accgaccaag
     5941 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6001 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6241 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6481 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6661 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     6781 gtccggaaat acaggaacgc acgctggatg gcccttcgct gggatggtga aaccatgaaa
     6841 aatggcagct tcagtggatt aagtgggggt aatgtggcct gtaccctctg gttgcatagg
     6901 tattcatacg gttaaaattt atcaggcgcg attgcggcag tttttcgggt ggtttgttgc
     6961 catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta
     7021 agggattatg gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta
     7081 ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
