LOCUS       sequence-423364-        6966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR5. C-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6966)
  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 6966)
  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_WDR5:1-334
FEATURES             Location/Qualifiers
     source          1..6966
                     /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             2141..2653
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2729..2747)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2784..2905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2795..2814)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2849..2868
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3150..3507
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3150..3341
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3173..3193)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3358..3493
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3538..4575
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4554..4562
                     /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    4599..4647
                     /note="synthetic polyadenylation signal"
     promoter        4803..4907
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4908..4976,4977..5768)
                     /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(5126..5145)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5926..6514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6006..6025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6630..6913
                     /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 acggaggaga agaagcccga gaccgaggcc gccagagcac agccaacccc
     1141 ttcgtcatcc gccactcaga gcaagcctac acctgtgaag ccaaactatg ctctaaagtt
     1201 cacccttgct ggccacacca aagcagtgtc ctccgtgaaa ttcagcccga atggagagtg
     1261 gctggcaagt tcatctgctg ataaacttat taaaatttgg ggcgcgtatg atgggaaatt
     1321 tgagaaaacc atatctggtc acaagctggg aatatccgat gtagcctggt cgtcagattc
     1381 taaccttctt gtttctgcct cagatgacaa aaccttgaag atatgggacg tgagctcggg
     1441 caagtgtctg aaaaccctga agggacacag taattatgtc ttttgctgca acttcaatcc
     1501 ccagtccaac cttattgtct caggatcctt tgacgaaagc gtgaggatat gggatgtgaa
     1561 aacagggaag tgcctcaaga ctttgccagc tcactcggat ccagtctcgg ccgttcattt
     1621 taatcgtgat ggatccttga tagtttcaag tagctatgat ggtctctgtc gcatctggga
     1681 caccgcctcg ggccagtgcc tgaagacgct catcgatgac gacaaccccc ccgtgtcttt
     1741 tgtgaagttc tccccgaacg gcaaatacat cctggccgcc acgctggaca acactctgaa
     1801 gctctgggac tacagcaagg ggaagtgcct gaagacgtac actggccaca agaatgagaa
     1861 atactgcata tttgccaatt tctctgttac tggtgggaag tggattgtgt ctggctcaga
     1921 ggataacctt gtttacatct ggaaccttca gacgaaagag attgtacaga aactacaagg
     1981 ccacacagat gtcgtgatct caacagcttg tcacccaaca gaaaacatca tcgcctctgc
     2041 tgcgctagaa aatgacaaaa caattaaact gtggaagagt gactgccgga tatctagatt
     2101 tgggcccaat tcctgcaggc gagctctcgg ctcgagcggc gtcttcacac tcgaagattt
     2161 cgttggggac tggcgacaga cagccggcta caacctggac caagtccttg aacagggagg
     2221 tgtgtccagt ttgtttcaga atctcggggt gtccgtaact ccgatccaaa ggattgtcct
     2281 gagcggtgaa aatgggctga agatcgacat ccatgtcatc atcccgtatg aaggtctgag
     2341 cggcgaccaa atgggccaga tcgaaaaaat ttttaaggtg gtgtaccctg tggatgatca
     2401 tcactttaag gtgatcctgc actatggcac actggtaatc gacggggtta cgccgaacat
     2461 gatcgactat ttcggacggc cgtatgaagg catcgccgtg ttcgacggca aaaagatcac
     2521 tgtaacaggg accctgtgga acggcaacaa aattatcgac gagcgcctga tcaaccccga
     2581 cggctccctg ctgttccgag taaccatcaa cggagtgacc ggctggcggc tgtgcgaacg
     2641 cattctggcg taaggccgcg actctagggc atgcaagctg atccggctgc taacaaagcc
     2701 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     2761 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2821 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2881 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2941 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     3001 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     3061 tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc
     3121 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     3181 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3241 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3301 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3361 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3421 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3481 ttttttggag gcctaggctt ttgcaaaaag ctcgattctt ctgacactag cgccaccatg
     3541 aagaagcccg aactcaccgc taccagcgtt gaaaaatttc tcatcgagaa gttcgacagt
     3601 gtgagcgacc tgatgcagtt gtcggagggc gaagagagcc gagccttcag cttcgatgtc
     3661 ggcggacgcg gctatgtact gcgggtgaat agctgcgctg atggcttcta caaagaccgc
     3721 tacgtgtacc gccacttcgc cagcgctgca ctacccatcc ccgaagtgtt ggacatcggc
     3781 gagttcagcg agagcctgac atactgcatc agtagacgcg cccaaggcgt tactctccaa
     3841 gacctccccg aaacagagct gcctgctgtg ttacagcctg tcgccgaagc tatggatgct
     3901 attgccgccg ccgacctcag tcaaaccagc ggcttcggcc cattcgggcc ccaaggcatc
     3961 ggccagtaca caacctggcg ggatttcatt tgcgccattg ctgatcccca tgtctaccac
     4021 tggcagaccg tgatggacga caccgtgtcc gccagcgtag ctcaagccct ggacgaactg
     4081 atgctgtggg ccgaagactg tcccgaggtg cgccacctcg tccatgccga cttcggcagc
     4141 aacaacgtcc tgaccgacaa cggccgcatc accgccgtaa tcgactggtc cgaagctatg
     4201 ttcggggaca gtcagtacga ggtggccaac atcttcttct ggcggccctg gctggcttgc
     4261 atggagcagc agactcgcta cttcgagcgc cggcatcccg agctggccgg cagccctcgt
     4321 ctgcgagcct acatgctgcg catcggcctg gatcagctct accagagcct cgtggacggc
     4381 aacttcgacg atgctgcctg ggctcaaggc cgctgcgatg ccatcgtccg cagcggggcc
     4441 ggcaccgtcg gtcgcacaca aatcgctcgc cggagcgcag ccgtatggac cgacggctgc
     4501 gtcgaggtgc tggccgacag cggcaaccgc cggcccagta cacgaccgcg cgctaaggag
     4561 gtaggtcgag tttagactct agccatcacg atggccgcaa taaaatatct ttattttcat
     4621 tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg
     4681 cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct
     4741 gcggactggc tttctacgta atggtttctt agacgtcagg tggcactttt cggggaaatg
     4801 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4861 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4921 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4981 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5041 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5101 caatgatgag cactttcaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5161 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5221 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5281 taaccatgag tgataacact gcggccaact tacttctgac aactatcgga ggaccgaagg
     5341 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5401 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5461 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5521 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5581 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5641 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5701 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5761 attggtaatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc
     5821 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5881 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5941 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6001 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6061 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6121 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6181 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6241 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6301 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6361 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6421 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6481 gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg
     6541 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6601 aaggatcttc acctagatcc ttttatagtc cggaaataca ggaacgcacg ctggatggcc
     6661 cttcgctggg atggtgaaac catgaaaaat ggcagcttca gtggattaag tgggggtaat
     6721 gtggcctgta ccctctggtt gcataggtat tcatacggtt aaaatttatc aggcgcgatt
     6781 gcggcagttt ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt gatcgcgctg
     6841 aacgcgtttt agcggtgcgt acaattaagg gattatggta aatccactta ctgtctgccc
     6901 tcgtagccat cgagataaac cgcagtactc cggccacgat gcgtccggcg tagaggatcg
     6961 agatct
//
