LOCUS       sequence-431699-        7335 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR41. C-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7335)
  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 7335)
  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_WDR41:1-459
FEATURES             Location/Qualifiers
     source          1..7335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        100..303
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     254..274
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          439..571
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     554..573
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     615..634
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        615..633
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2092..2604
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2680..2698)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2735..2856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2746..2765)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2800..2819
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3101..3458
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3101..3292
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3124..3144)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3309..3444
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3489..4526
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4505..4513
                     /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    4550..4598
                     /note="synthetic polyadenylation signal"
     promoter        4754..4858
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4859..4927,4928..5719)
                     /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(5077..5096)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5877..6465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5957..5976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6581..6864
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        join(7055..7335,1..99)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
       61 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      121 accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      181 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac
      241 cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      301 gctggtttag tgaaccgtca gatcactaga agctttattg cggtagttta tcacagttaa
      361 attgctaacg cagtcagtgc ttctgacaca acagtctcga acttaagctg cagaagttgg
      421 tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag
      481 aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt
      541 actgacatcc actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct
      601 taaggctaga gtattaatac gactcactat agggctagca aagcgatcgc ttccgaattc
      661 gccaccatgt tgcgatggct gatcggggga ggccgagaac cgcagggact ggccgagaaa
      721 tctcctttac agacaatagg tgaagaacaa acccagaatc cctacactga actgctagta
      781 ctgaaggctc atcatgatat tgtacgattt ctggtacagt tagatgacta cagatttgca
      841 tctgctggtg atgatggaat tgtagttgtg tggaatgccc agacagggga aaaactttta
      901 gaactgaatg gacacactca aaagataaca gctattatta catttccttc cttggaatct
      961 tgtgaagaga aaaatcaact catcttgaca gcctctgctg atagaacagt tattgtgtgg
     1021 gatggtgata ctaccagaca agttcagaga atatcatgct tccagtctac tgtaaagtgt
     1081 ttaactgttc ttcagagact agatgtttgg ctttctggtg ggaatgacct gtgtgtgtgg
     1141 aaccgaaaat tagatctcct gtgtaagact agccaccttt ctgatacagg tattagtgct
     1201 ttggttgaaa tacctaagaa ctgtgttgtg gcagcagttg gcaaagaact gataattttc
     1261 aggttggtag cacccacaga aggatcacta gaatgggata ttcttgaagt taagcgcctc
     1321 cttgatcacc aggataatat tctctcattg attaatgtca atgatttgag ttttgtcacc
     1381 ggctcccacg tcggagagct gatcatctgg gatgccctgg actggaccat gcaggcctat
     1441 gaacgcaact tctgggaccc atctccacaa ctggacaccc aacaagaaat aaaactctgt
     1501 caaaaatcaa atgacatttc tattcatcat ttcacatgtg atgaagagaa tgtatttgct
     1561 gcagttggaa ggggtttata cgtgtatagc cttcaaatga agcgtgtgat tgcctgccag
     1621 aaaactgcac atgactccaa tgtcctgcac gttgccagac ttccaaacag gcagttaatc
     1681 tcatgctcag aagatggcag tgtacgcatt tgggagttaa gagaaaaaca gcagcttgca
     1741 gctgagcctg taccaacagg tttttttaac atgtggggat ttggaagagt cagcaaacaa
     1801 gccagccaac ctgttaaaaa gcagcaagaa aatgctactt catgttcact ggagcttatt
     1861 ggagatttga ttggacactc atcatctgtg gagatgtttc tatactttga agatcatgga
     1921 ctagtgacgt gctccgctga tcatctcatt attttgtgga aaaatggaga gcgagaatct
     1981 ggattgcgca gtttaagatt atttcaaaaa ttagaggaga atggtgactt ataccttgct
     2041 gtctctagat ttgggcccaa ttcctgcagg cgagctctcg gctcgagcgg cgtcttcaca
     2101 ctcgaagatt tcgttgggga ctggcgacag acagccggct acaacctgga ccaagtcctt
     2161 gaacagggag gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac tccgatccaa
     2221 aggattgtcc tgagcggtga aaatgggctg aagatcgaca tccatgtcat catcccgtat
     2281 gaaggtctga gcggcgacca aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct
     2341 gtggatgatc atcactttaa ggtgatcctg cactatggca cactggtaat cgacggggtt
     2401 acgccgaaca tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc
     2461 aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca aaattatcga cgagcgcctg
     2521 atcaaccccg acggctccct gctgttccga gtaaccatca acggagtgac cggctggcgg
     2581 ctgtgcgaac gcattctggc gtaaggccgc gactctaggg catgcaagct gatccggctg
     2641 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     2701 aaccccttgg ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     2761 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2821 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2881 atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa
     2941 tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     3001 ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc
     3061 ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc
     3121 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3181 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3241 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3301 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3361 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3421 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgattct tctgacacta
     3481 gcgccaccat gaagaagccc gaactcaccg ctaccagcgt tgaaaaattt ctcatcgaga
     3541 agttcgacag tgtgagcgac ctgatgcagt tgtcggaggg cgaagagagc cgagccttca
     3601 gcttcgatgt cggcggacgc ggctatgtac tgcgggtgaa tagctgcgct gatggcttct
     3661 acaaagaccg ctacgtgtac cgccacttcg ccagcgctgc actacccatc cccgaagtgt
     3721 tggacatcgg cgagttcagc gagagcctga catactgcat cagtagacgc gcccaaggcg
     3781 ttactctcca agacctcccc gaaacagagc tgcctgctgt gttacagcct gtcgccgaag
     3841 ctatggatgc tattgccgcc gccgacctca gtcaaaccag cggcttcggc ccattcgggc
     3901 cccaaggcat cggccagtac acaacctggc gggatttcat ttgcgccatt gctgatcccc
     3961 atgtctacca ctggcagacc gtgatggacg acaccgtgtc cgccagcgta gctcaagccc
     4021 tggacgaact gatgctgtgg gccgaagact gtcccgaggt gcgccacctc gtccatgccg
     4081 acttcggcag caacaacgtc ctgaccgaca acggccgcat caccgccgta atcgactggt
     4141 ccgaagctat gttcggggac agtcagtacg aggtggccaa catcttcttc tggcggccct
     4201 ggctggcttg catggagcag cagactcgct acttcgagcg ccggcatccc gagctggccg
     4261 gcagccctcg tctgcgagcc tacatgctgc gcatcggcct ggatcagctc taccagagcc
     4321 tcgtggacgg caacttcgac gatgctgcct gggctcaagg ccgctgcgat gccatcgtcc
     4381 gcagcggggc cggcaccgtc ggtcgcacac aaatcgctcg ccggagcgca gccgtatgga
     4441 ccgacggctg cgtcgaggtg ctggccgaca gcggcaaccg ccggcccagt acacgaccgc
     4501 gcgctaagga ggtaggtcga gtttagactc tagccatcac gatggccgca ataaaatatc
     4561 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatcct
     4621 ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca
     4681 gcaccgtttc tgcggactgg ctttctacgt aatggtttct tagacgtcag gtggcacttt
     4741 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4801 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4861 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4921 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4981 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5041 agaacgtttt ccaatgatga gcactttcaa agttctgcta tgtggcgcgg tattatcccg
     5101 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5161 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5221 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caactatcgg
     5281 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5341 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5401 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5461 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5521 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5581 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5641 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5701 actgattaag cattggtaat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct
     5761 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5821 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5881 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5941 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6001 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6061 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6121 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6181 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6241 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6301 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6361 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6421 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc
     6481 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6541 agattatcaa aaaggatctt cacctagatc cttttatagt ccggaaatac aggaacgcac
     6601 gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa
     6661 gtgggggtaa tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat
     6721 caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg
     6781 tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt
     6841 actgtctgcc ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc
     6901 gtagaggatc gagatcttca atattggcca ttagccatat tattcattgg ttatatagca
     6961 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
     7021 tatattggct catgtccaat atgaccgcca tgttggcatt gattattgac tagttattaa
     7081 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     7141 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     7201 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     7261 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
     7321 cctattgacg tcaat
//
