LOCUS       sequence-423334-        8372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human WDR91. N-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8372)
  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 8372)
  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: pHTN-HaloTag_WDR91:1-747
FEATURES             Location/Qualifiers
     source          1..8372
                     /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      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1957
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(4323..4341)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4373..4494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4384..4403)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4438..4457
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4739..5096
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4739..4930
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4762..4782)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4947..5082
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5147..5941
                     /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(5201..5220)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5811..5830
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6005..6053
                     /note="synthetic polyadenylation signal"
     promoter        6209..6313
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6314..6382,6383..7174)
                     /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(6532..6551)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7332..7920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7412..7431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8036..8319
                     /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 gccaccatgg cagaaatcgg
     1081 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1141 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1201 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1261 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1321 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1381 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1441 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1501 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1561 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1621 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1681 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1741 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1801 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1861 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1921 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccactg aggatctgta
     1981 ctttcagagc gataacgcga tcgcttccga attcatggcg gaggccgtgg agcgcactga
     2041 cgagctggtc cgggagtacc tgctcttccg cgggttcacg cacacactgc ggcagctgga
     2101 cgccgagatc aaggcggaca aggagaaggg gttccgggtg gataagattg tggaccagct
     2161 gcagcagtta atgcaggtgt atgacttggc tgcccttcgg gattattgga gctacttgga
     2221 gcgtcggctc ttcagccgct tggaggatat atacagaccc acaatccaca agctgaaaac
     2281 cagcctgttt cgattttatc ttgtctacac aatccagaca aacagaaatg acaaggctca
     2341 ggagttcttt gcaaagcagg ccacggaact ccagaaccag gctgagtgga aggattggtt
     2401 tgtcctgccc ttcctgccat ccccggacac caaccccacc tttgctacct acttttctcg
     2461 acagtgggct gacaccttca ttgtgtccct gcacaacttc ctgagcgtcc tgtttcagtg
     2521 catgccagtc cctgtgatcc tgaactttga tgcggagtgt cagaggacta accaggttca
     2581 agaagaaaat gaagttctgc gtcagaagct ttttgcattg caagctgaaa tccaccgact
     2641 gaagaaagag gagcaacagc cagaagagga agaggccttg gtccaacaca aattgcctcc
     2701 ttatgtctcc aacatggacc gcctggggga ctcggaactt gccatggtgt gcagccaaag
     2761 gaatgcctcc ctctcccagt cacctcgtgt gggcttcctg tcctcgctgc tgcctcagag
     2821 taagaagagc ccctcaaggt tgtcgcctgc tcagggccct cctcaacctc agagctcggc
     2881 caagaaagag tccttcggtg gtcagggcac caagggaaag gacccgacgt ccggagccaa
     2941 ggatgggaag agcctcctca gcgggctggc cactggggag tccggttggt cacagcaccg
     3001 gcagcggcgc ctgcaggacc atggcaagga gaggaaggag cttttctcca caaccacttc
     3061 ccagtgtgca gagaagaaac cagaagccag tggcccagag gctgagccct gcccagagct
     3121 ccacacggag ccagtggagc cactgactcg ggcatcctcg gcaggccctg agggtggagg
     3181 agtccgcccc gagcagccct ttattgtgct gggacaggag gagtacgggg aacaccactc
     3241 atccatcatg cactgcagag tggactgctc tgggaggaga gtcgccagct tagacgtaga
     3301 tggggtcatc aaagtgtggt ccttcaaccc catcatgcag accaaagcat cctccatttc
     3361 caaatcaccg ctgctgtctt tggaatgggc caccaaacgg gacagactgc tcttgctggg
     3421 cagtggtgtg ggaacagtgc gtctctatga cacggaagcc aagaagaatc tctgtgaaat
     3481 caatatcaac gacaacatgc ccagaatcct gtctcttgcg tgcagcccca acggggcctc
     3541 tttcgtctgt tcggcagcag ctccgagcct cacttcccag gtggacttct cagcaccaga
     3601 catcggcagc aagggcatga accaggttcc tggcaggctg ctgctgtggg acacgaaaac
     3661 catgaagcag cagctccagt tctccctgga tccagaaccc attgctatca actgtacagc
     3721 cttcaatcac aacgggaacc tgctggtcac aggggcagct gatggcgtca tccggctgtt
     3781 tgacatgcag cagcatgagt gcgcgatgag ctggagggcc cactacgggg aggtctactc
     3841 tgtggagttc agctatgatg agaacaccgt gtacagcatc ggcgaggacg ggaagttcat
     3901 ccagtggaac atccacaaga gtggcctcaa ggtatccgag tacagcctcc cctcagatgc
     3961 cacgggcccc tttgtgctgt ctggatacag cggctacaag caggttcaag tccccagggg
     4021 ccgactcttc gcttttgact cggagggaaa ttacatgctg acatgttctg ccacaggcgg
     4081 cgtcatctac aagctgggtg gcgatgagaa ggttctggag agctgcttga gcctaggtgg
     4141 ccaccgagcc cctgtggtca ccgtggactg gagcactgcc atggactgtg ggacctgcct
     4201 caccgcctcc atggatggca agatcaagct gaccaccctc ctggcccata aagcctgatg
     4261 atctagattt gggcccaatt cctgcaggat tttgcggccg cttgctgagt tggctgctgc
     4321 caccgctgag caataactag cataacccct tggccgcttc gagcagacat gataagatac
     4381 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     4441 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4501 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc
     4561 aagtaaaacc tctacaaatg tggtaaaatc gaattttaac aaaatattaa cgcttacaat
     4621 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgcgga
     4681 tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct
     4741 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4801 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4861 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4921 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4981 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     5041 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5101 ttgattcttc tgacacaaca gtctcgaact taaggctaga gccaccatga ttgaacaaga
     5161 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     5221 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5281 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     5341 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5401 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5461 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5521 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5581 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5641 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     5701 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5761 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5821 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5881 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5941 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg
     6001 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
     6061 agcgataagg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac
     6121 attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtaatgg tttcttagac
     6181 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6241 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6301 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6361 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6421 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6481 gagttttcgc cccgaagaac gttttccaat gatgagcact ttcaaagttc tgctatgtgg
     6541 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6601 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6661 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6721 tctgacaact atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6781 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6841 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6901 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6961 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7021 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7081 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7141 tgagataggt gcctcactga ttaagcattg gtaattcgaa atgaccgacc aagcgacgcc
     7201 caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7261 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7321 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7381 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7441 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7501 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7561 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7621 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7681 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7741 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7801 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7861 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa
     7921 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7981 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga
     8041 aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg aaaaatggca
     8101 gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat aggtattcat
     8161 acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt tgccattttt
     8221 acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa ttaagggatt
     8281 atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca gtactccggc
     8341 cacgatgcgt ccggcgtaga ggatcgagat ct
//
