LOCUS       sequence-423363-        6910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR5. N-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6910)
  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 6910)
  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_WDR5:1-334
FEATURES             Location/Qualifiers
     source          1..6910
                     /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(2678..2696)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2728..2849
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2739..2758)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2793..2812
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3094..3451
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3094..3285
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3117..3137)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3302..3437
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3482..4519
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4498..4506
                     /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    4543..4591
                     /note="synthetic polyadenylation signal"
     promoter        4747..4851
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4852..4920,4921..5712)
                     /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(5070..5089)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5870..6458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5950..5969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6574..6857
                     /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 gcgacggagg
     1621 agaagaagcc cgagaccgag gccgccagag cacagccaac cccttcgtca tccgccactc
     1681 agagcaagcc tacacctgtg aagccaaact atgctctaaa gttcaccctt gctggccaca
     1741 ccaaagcagt gtcctccgtg aaattcagcc cgaatggaga gtggctggca agttcatctg
     1801 ctgataaact tattaaaatt tggggcgcgt atgatgggaa atttgagaaa accatatctg
     1861 gtcacaagct gggaatatcc gatgtagcct ggtcgtcaga ttctaacctt cttgtttctg
     1921 cctcagatga caaaaccttg aagatatggg acgtgagctc gggcaagtgt ctgaaaaccc
     1981 tgaagggaca cagtaattat gtcttttgct gcaacttcaa tccccagtcc aaccttattg
     2041 tctcaggatc ctttgacgaa agcgtgagga tatgggatgt gaaaacaggg aagtgcctca
     2101 agactttgcc agctcactcg gatccagtct cggccgttca ttttaatcgt gatggatcct
     2161 tgatagtttc aagtagctat gatggtctct gtcgcatctg ggacaccgcc tcgggccagt
     2221 gcctgaagac gctcatcgat gacgacaacc cccccgtgtc ttttgtgaag ttctccccga
     2281 acggcaaata catcctggcc gccacgctgg acaacactct gaagctctgg gactacagca
     2341 aggggaagtg cctgaagacg tacactggcc acaagaatga gaaatactgc atatttgcca
     2401 atttctctgt tactggtggg aagtggattg tgtctggctc agaggataac cttgtttaca
     2461 tctggaacct tcagacgaaa gagattgtac agaaactaca aggccacaca gatgtcgtga
     2521 tctcaacagc ttgtcaccca acagaaaaca tcatcgcctc tgctgcgcta gaaaatgaca
     2581 aaacaattaa actgtggaag agtgactgct aagatatcta gatttgggcc caattcctgc
     2641 aggattttgc ggccgcttgc tgagttggct gctgccaccg ctgagcaata actagcataa
     2701 ccccttggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     2761 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2821 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2881 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     2941 aaatcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     3001 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     3061 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     3121 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3181 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3241 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3301 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3361 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3421 aggctttttt ggaggcctag gcttttgcaa aaagctcgat tcttctgaca ctagcgccac
     3481 catgaagaag cccgaactca ccgctaccag cgttgaaaaa tttctcatcg agaagttcga
     3541 cagtgtgagc gacctgatgc agttgtcgga gggcgaagag agccgagcct tcagcttcga
     3601 tgtcggcgga cgcggctatg tactgcgggt gaatagctgc gctgatggct tctacaaaga
     3661 ccgctacgtg taccgccact tcgccagcgc tgcactaccc atccccgaag tgttggacat
     3721 cggcgagttc agcgagagcc tgacatactg catcagtaga cgcgcccaag gcgttactct
     3781 ccaagacctc cccgaaacag agctgcctgc tgtgttacag cctgtcgccg aagctatgga
     3841 tgctattgcc gccgccgacc tcagtcaaac cagcggcttc ggcccattcg ggccccaagg
     3901 catcggccag tacacaacct ggcgggattt catttgcgcc attgctgatc cccatgtcta
     3961 ccactggcag accgtgatgg acgacaccgt gtccgccagc gtagctcaag ccctggacga
     4021 actgatgctg tgggccgaag actgtcccga ggtgcgccac ctcgtccatg ccgacttcgg
     4081 cagcaacaac gtcctgaccg acaacggccg catcaccgcc gtaatcgact ggtccgaagc
     4141 tatgttcggg gacagtcagt acgaggtggc caacatcttc ttctggcggc cctggctggc
     4201 ttgcatggag cagcagactc gctacttcga gcgccggcat cccgagctgg ccggcagccc
     4261 tcgtctgcga gcctacatgc tgcgcatcgg cctggatcag ctctaccaga gcctcgtgga
     4321 cggcaacttc gacgatgctg cctgggctca aggccgctgc gatgccatcg tccgcagcgg
     4381 ggccggcacc gtcggtcgca cacaaatcgc tcgccggagc gcagccgtat ggaccgacgg
     4441 ctgcgtcgag gtgctggccg acagcggcaa ccgccggccc agtacacgac cgcgcgctaa
     4501 ggaggtaggt cgagtttaga ctctagccat cacgatggcc gcaataaaat atctttattt
     4561 tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat cctctttgcg
     4621 cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt
     4681 ttctgcggac tggctttcta cgtaatggtt tcttagacgt caggtggcac ttttcgggga
     4741 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4801 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4861 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4921 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4981 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5041 tttccaatga tgagcacttt caaagttctg ctatgtggcg cggtattatc ccgtattgac
     5101 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5161 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5221 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaactat cggaggaccg
     5281 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5341 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5401 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5461 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5521 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5581 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5641 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5701 aagcattggt aattcgaaat gaccgaccaa gcgacgccca accggtatca gctcactcaa
     5761 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5821 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5881 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5941 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6001 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6061 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6121 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6181 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6241 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6301 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6361 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6421 gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta
     6481 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6541 caaaaaggat cttcacctag atccttttat agtccggaaa tacaggaacg cacgctggat
     6601 ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc ttcagtggat taagtggggg
     6661 taatgtggcc tgtaccctct ggttgcatag gtattcatac ggttaaaatt tatcaggcgc
     6721 gattgcggca gtttttcggg tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc
     6781 gctgaacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct
     6841 gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg
     6901 atcgagatct
//
