LOCUS       sequence-426277-        6407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of human WDR5 fragment A2 to C334. N-terminal 
            10X His tag with TEV cleavage and C-terminal biotin acceptor peptide
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6407)
  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 6407)
  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: pNICBio2_WDR5:2-334
FEATURES             Location/Qualifiers
     source          1..6407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(51..68)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(121..165)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(1186..1206)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(1219..1248)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             1259..1281
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1296..1320
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(1320..1339)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1321..1339)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1423..1442)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1648..1725
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1726..2808
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYQLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1745..1764)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3617..3808
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3802..3824)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3910..4049
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3965..3984
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4064..4081)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4235..4823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4315..4334)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4934..4953)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4945..5760
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4945..5760
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5018..5037)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5852..6307)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5989..6010)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6201..6220
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(6344..6391)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     6387..6405
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 gcagcagcca actcagcttc ctttcgggct ttgttagcag ccggatctca gtggtggtgg
       61 tggtggtgct cgagtgcggc cgcaagcttg tcgacggagc tcgaattcgg atccttatca
      121 ttcatgccat tcaatcttct gcgcttcaaa gatatcattc aggccgtatc cacctttact
      181 gctgcagtca ctcttccaca gtttaattgt tttgtcattt tctagcgcag cagaggcgat
      241 gatgttttct gttgggtgac aagctgttga gatcacgaca tctgtgtggc cttgtagttt
      301 ctgtacaatc tctttcgtct gaaggttcca gatgtaaaca aggttatcct ctgagccaga
      361 cacaatccac ttcccaccag taacagagaa attggcaaat atgcagtatt tctcattctt
      421 gtggccagtg tacgtcttca ggcacttccc cttgctgtag tcccagagct tcagagtgtt
      481 gtccagcgtg gcggccagga tgtatttgcc gttcggggag aacttcacaa aagacacggg
      541 ggggttgtcg tcatcgatga gcgtcttcag gcactggccc gaggcggtgt cccagatgcg
      601 acagagacca tcatagctac ttgaaactat caaggatcca tcacgattaa aatgaacggc
      661 cgagactgga tccgagtgag ctggcaaagt cttgaggcac ttccctgttt tcacatccca
      721 tatcctcacg ctttcgtcaa aggatcctga gacaataagg ttggactggg gattgaagtt
      781 gcagcaaaag acataattac tgtgtccctt cagggttttc agacacttgc ccgagctcac
      841 gtcccatatc ttcaaggttt tgtcatctga ggcagaaaca agaaggttag aatctgacga
      901 ccaggctaca tcggatattc ccagcttgtg accagatatg gttttctcaa atttcccatc
      961 atacgcgccc caaattttaa taagtttatc agcagatgaa cttgccagcc actctccatt
     1021 cgggctgaat ttcacggagg acactgcttt ggtgtggcca gcaagggtga actttagagc
     1081 atagtttggc ttcacaggtg taggcttgct ctgagtggcg gatgacgaag gggttggctg
     1141 tgctctggcg gcctcggtct cgggcttctt ctcctccgtc gccatggatt ggaagtacag
     1201 gttctcggta cccagatcgt gatgatggtg atgatgatga tgatggtgca tatgtatatc
     1261 tccttcttaa agttaaacaa aattatttct agaggggaat tgttatccgc tcacaattcc
     1321 cctatagtga gtcgtattaa tttcgcggga tcgagatctc gatcctctac gccggacgca
     1381 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
     1441 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
     1501 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
     1561 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt
     1621 cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt
     1681 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     1741 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     1801 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     1861 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     1921 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     1981 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     2041 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     2101 tatcagctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     2161 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     2221 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     2281 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     2341 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     2401 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     2461 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     2521 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     2581 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     2641 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     2701 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     2761 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     2821 taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt
     2881 caacccagtc agctccttcc ggtgggcgcg ggacatgact atcgtcgccg cacttatgac
     2941 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg
     3001 cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat
     3061 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa
     3121 gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc tcctgtcgtt
     3181 gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg
     3241 cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat
     3301 ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat
     3361 tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt
     3421 attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac
     3481 cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg
     3541 tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc
     3601 ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat
     3661 cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc
     3721 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg
     3781 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     3841 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3901 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     3961 tatgcggcat cagagcagat tgtactgaga gtgcaccatt gcggtgtgaa ataccgcaca
     4021 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
     4081 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4141 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4201 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4261 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4321 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4381 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4441 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4501 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4561 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4621 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     4681 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4741 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4801 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4861 agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact gtctgcttac
     4921 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc ttgctctagg
     4981 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     5041 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg
     5101 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     5161 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     5221 gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa
     5281 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     5341 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     5401 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     5461 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     5521 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     5581 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     5641 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     5701 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     5761 gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5821 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     5881 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5941 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     6001 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     6061 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     6121 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     6181 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     6241 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6301 ggcgcgtccc attcgccaat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga
     6361 cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtg
//
