LOCUS       sequence-423335-        8384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR91. C-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8384)
  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 8384)
  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: pHTC-HaloTag_WDR91:1-747
FEATURES             Location/Qualifiers
     source          1..8384
                     /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             3338..3358
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             3374..4258
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(4330..4348)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4385..4506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4396..4415)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4450..4469
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4751..5108
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4751..4942
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4774..4794)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4959..5094
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5159..5953
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5213..5232)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5823..5842
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6017..6065
                     /note="synthetic polyadenylation signal"
     promoter        6221..6325
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6326..6394,6395..7186)
                     /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(6544..6563)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7344..7932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7424..7443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8048..8331
                     /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 ccgaattcac
     1081 catggcggag gccgtggagc gcactgacga gctggtccgg gagtacctgc tcttccgcgg
     1141 gttcacgcac acactgcggc agctggacgc cgagatcaag gcggacaagg agaaggggtt
     1201 ccgggtggat aagattgtgg accagctgca gcagttaatg caggtgtatg acttggctgc
     1261 ccttcgggat tattggagct acttggagcg tcggctcttc agccgcttgg aggatatata
     1321 cagacccaca atccacaagc tgaaaaccag cctgtttcga ttttatcttg tctacacaat
     1381 ccagacaaac agaaatgaca aggctcagga gttctttgca aagcaggcca cggaactcca
     1441 gaaccaggct gagtggaagg attggtttgt cctgcccttc ctgccatccc cggacaccaa
     1501 ccccaccttt gctacctact tttctcgaca gtgggctgac accttcattg tgtccctgca
     1561 caacttcctg agcgtcctgt ttcagtgcat gccagtccct gtgatcctga actttgatgc
     1621 ggagtgtcag aggactaacc aggttcaaga agaaaatgaa gttctgcgtc agaagctttt
     1681 tgcattgcaa gctgaaatcc accgactgaa gaaagaggag caacagccag aagaggaaga
     1741 ggccttggtc caacacaaat tgcctcctta tgtctccaac atggaccgcc tgggggactc
     1801 ggaacttgcc atggtgtgca gccaaaggaa tgcctccctc tcccagtcac ctcgtgtggg
     1861 cttcctgtcc tcgctgctgc ctcagagtaa gaagagcccc tcaaggttgt cgcctgctca
     1921 gggccctcct caacctcaga gctcggccaa gaaagagtcc ttcggtggtc agggcaccaa
     1981 gggaaaggac ccgacgtccg gagccaagga tgggaagagc ctcctcagcg ggctggccac
     2041 tggggagtcc ggttggtcac agcaccggca gcggcgcctg caggaccatg gcaaggagag
     2101 gaaggagctt ttctccacaa ccacttccca gtgtgcagag aagaaaccag aagccagtgg
     2161 cccagaggct gagccctgcc cagagctcca cacggagcca gtggagccac tgactcgggc
     2221 atcctcggca ggccctgagg gtggaggagt ccgccccgag cagcccttta ttgtgctggg
     2281 acaggaggag tacggggaac accactcatc catcatgcac tgcagagtgg actgctctgg
     2341 gaggagagtc gccagcttag acgtagatgg ggtcatcaaa gtgtggtcct tcaaccccat
     2401 catgcagacc aaagcatcct ccatttccaa atcaccgctg ctgtctttgg aatgggccac
     2461 caaacgggac agactgctct tgctgggcag tggtgtggga acagtgcgtc tctatgacac
     2521 ggaagccaag aagaatctct gtgaaatcaa tatcaacgac aacatgccca gaatcctgtc
     2581 tcttgcgtgc agccccaacg gggcctcttt cgtctgttcg gcagcagctc cgagcctcac
     2641 ttcccaggtg gacttctcag caccagacat cggcagcaag ggcatgaacc aggttcctgg
     2701 caggctgctg ctgtgggaca cgaaaaccat gaagcagcag ctccagttct ccctggatcc
     2761 agaacccatt gctatcaact gtacagcctt caatcacaac gggaacctgc tggtcacagg
     2821 ggcagctgat ggcgtcatcc ggctgtttga catgcagcag catgagtgcg cgatgagctg
     2881 gagggcccac tacggggagg tctactctgt ggagttcagc tatgatgaga acaccgtgta
     2941 cagcatcggc gaggacggga agttcatcca gtggaacatc cacaagagtg gcctcaaggt
     3001 atccgagtac agcctcccct cagatgccac gggccccttt gtgctgtctg gatacagcgg
     3061 ctacaagcag gttcaagtcc ccaggggccg actcttcgct tttgactcgg agggaaatta
     3121 catgctgaca tgttctgcca caggcggcgt catctacaag ctgggtggcg atgagaaggt
     3181 tctggagagc tgcttgagcc taggtggcca ccgagcccct gtggtcaccg tggactggag
     3241 cactgccatg gactgtggga cctgcctcac cgcctccatg gatggcaaga tcaagctgac
     3301 caccctcctg gcccataaag ccctcgagcc aaccactgag gatctgtact ttcagagcga
     3361 taacgatgga tccgaaatcg gtactggctt tccattcgac ccccattatg tggaagtcct
     3421 gggcgagcgc atgcactacg tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct
     3481 gcacggtaac ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac
     3541 ccatcgctgc attgctccag acctgatcgg tatgggcaaa tccgacaaac cagacctggg
     3601 ttatttcttc gacgaccacg tccgcttcat ggatgccttc atcgaagccc tgggtctgga
     3661 agaggtcgtc ctggtcattc acgactgggg ctccgctctg ggtttccact gggccaagcg
     3721 caatccagag cgcgtcaaag gtattgcatt tatggagttc atccgcccta tcccgacctg
     3781 ggacgaatgg ccagaatttg cccgcgagac cttccaggcc ttccgcacca ccgacgtcgg
     3841 ccgcaagctg atcatcgatc agaacgtttt tatcgagggt acgctgccga tgggtgtcgt
     3901 ccgcccgctg actgaagtcg agatggacca ttaccgcgag ccgttcctga atcctgttga
     3961 ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc gccggtgagc cagcgaacat
     4021 cgtcgcgctg gtcgaagaat acatggactg gctgcaccag tcccctgtcc cgaagctgct
     4081 gttctggggc accccaggcg ttctgatccc accggccgaa gccgctcgcc tggccaaaag
     4141 cctgcctaac tgcaaggctg tggacatcgg cccgggtctg aatctgctgc aagaagacaa
     4201 cccggacctg atcggcagcg agatcgcgcg ctggctgtct actctggaga tttccggtta
     4261 atagaattgg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     4321 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct tcgagcagac
     4381 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     4441 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     4501 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     4561 gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt
     4621 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4681 cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     4741 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4801 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4861 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4921 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4981 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5041 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5101 tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaaggcta gagccaccat
     5161 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5221 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5281 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5341 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5401 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5461 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5521 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5581 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5641 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5701 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5761 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5821 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5881 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5941 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     6001 ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     6061 gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg ttttcccttg tccagatagc
     6121 ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtaat
     6181 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     6241 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6301 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6361 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6421 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6481 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca ctttcaaagt
     6541 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6601 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6661 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6721 ggccaactta cttctgacaa ctatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6781 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6841 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6901 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6961 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     7021 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     7081 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     7141 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaattcg aaatgaccga
     7201 ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7261 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7321 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7381 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7441 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7501 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7561 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7621 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7681 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7741 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7801 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7861 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7921 aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7981 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     8041 ttatagtccg gaaatacagg aacgcacgct ggatggccct tcgctgggat ggtgaaacca
     8101 tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc
     8161 ataggtattc atacggttaa aatttatcag gcgcgattgc ggcagttttt cgggtggttt
     8221 gttgccattt ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac
     8281 aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg agataaaccg
     8341 cagtactccg gccacgatgc gtccggcgta gaggatcgag atct
//
