LOCUS       sequence-423342-        8370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human BOP1. N-terminal HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8370)
  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 8370)
  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_BOP1:1-746
FEATURES             Location/Qualifiers
     source          1..8370
                     /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(4321..4339)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4371..4492
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4382..4401)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4436..4455
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4737..5094
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4737..4928
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4760..4780)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4945..5080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5145..5939
                     /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(5199..5218)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5809..5828
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6003..6051
                     /note="synthetic polyadenylation signal"
     promoter        6207..6311
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6312..6380,6381..7172)
                     /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(6530..6549)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7330..7918)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7410..7429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8034..8317
                     /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 ggttcgcggg gtgcggggcg
     2041 cacggcggcg ccgagcgtgc ggccggagaa gcggcggtct gagcccgaac tggagcctga
     2101 gcccgagccg gagccccccc tcctctgcac ctctcctctc agccacagca ccggcagcga
     2161 ttctggcgtc tccgacagcg aggagagtgt gttctcaggc ctggaagatt ccggcagtga
     2221 cagcagtgag gatgatgacg aaggcgacga ggagggagag gacggagccc ttgatgacga
     2281 gggccacagt gggattaaaa agaccactga ggagcaggtg caggccagca ctccttgccc
     2341 gaggacagag atggcgagcg cccggattgg ggatgagtat gcggaggaca gctctgatga
     2401 ggaggacatc cggaacacgg tgggcaacgt gcccttggag tggtacgatg acttccccca
     2461 cgtgggctac gacctggatg gcaggcgcat ctacaagccc ctgcggaccc gggatgagct
     2521 ggaccagttc ctggacaaga tggacgatcc tgactactgg cgcaccgtgc aggacccgat
     2581 gacagggcgg gacctgagac tgacggatga gcaggtggcc ctggtgcggc ggctgcagag
     2641 tggccagttt ggggatgtgg gcttcaaccc ctatgagccg gctgtcgact tcttcagcgg
     2701 ggacgtcatg atccacccgg tgaccaaccg cccggccgac aagcgcagct tcatcccctc
     2761 cctggtggag aaggagaagg tctctcgcat ggtgcacgcc atcaagatgg gctggatcca
     2821 gcctcgccgg ccccgagacc ccacccccag cttctatgac ctgtgggccc aggaggaccc
     2881 caacgccgtg ctcgggcgcc acaagatgca cgtacctgct cccaagctgg ccctgccagg
     2941 ccacgccgag tcgtacaacc caccccctga atacctgctc agcgaggagg agcgcttggc
     3001 gtgggaacag caggagccag gcgagaggaa gctgagcttt ttgccacgca agttcccgag
     3061 cctgcgggcc gtgcctgcct acggacgctt catccaggaa cgcttcgagc gctgccttga
     3121 cctgtacctg tgcccacggc agcgcaagat gagggtgaat gtagaccctg aggacctcat
     3181 ccccaagctg cctcggccga gggacctgca gcccttcccc acgtgccagg ccctggtcta
     3241 caggggccac agtgaccttg tccggtgcct cagtgtctct cctgggggcc agtggctggt
     3301 ttcaggctct gacgacggct ccctgcggct ctgggaggtg gccactgccc gctgtgtgag
     3361 gactgttccc gtggggggcg tggtgaagag tgtggcctgg aaccccagcc ccgctgtctg
     3421 cctggtggct gcagccgtgg aggactcggt gctgctgctg aacccagctc tgggggaccg
     3481 gctggtggcg ggcagcacag atcagctgtt gagcgccttc gtcccgcctg aggagccccc
     3541 cttgcagccg gcccgctggc tggaggcctc agaggaggag cgccaagtgg gcctgcggct
     3601 gcgcatctgc cacgggaagc cagtgacgca ggtgacctgg cacgggcgtg gggactacct
     3661 ggccgtggtg ctggccaccc aaggccacac ccaggtgctg attcaccagc tgagccgtcg
     3721 ccgcagccag agtccgttcc gccgcagcca cggacaggtg cagcgagtgg ccttccaccc
     3781 tgcccggccc ttcctgttgg tggcgtccca gcgcagcgtc cgcctctacc acctgctgcg
     3841 ccaggagctc accaagaagc tgatgcccaa ctgcaagtgg gtgtccagcc tggcggtgca
     3901 ccctgcaggt gacaacgtca tctgtgggag ctacgatagc aagctggtgt ggtttgacct
     3961 ggatctttcc accaagccat acaggatgct gagacaccac aagaaggctc tgcgggctgt
     4021 ggccttccac ccgcggtacc cactctttgc gtcaggctcg gacgacggca gtgtcatcgt
     4081 ctgccatggc atggtgtaca atgaccttct gcagaacccc ttgctggtgc ccgtcaaggt
     4141 gctgaaggga cacgtgctga cccgagatct gggagtgctg gacgtcatct tccaccccac
     4201 ccagccgtgg gtcttctcct cgggggcaga cgggactgtc cgcctcttca cctaggatat
     4261 ctagatttgg gcccaattcc tgcaggattt tgcggccgct tgctgagttg gctgctgcca
     4321 ccgctgagca ataactagca taaccccttg gccgcttcga gcagacatga taagatacat
     4381 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     4441 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4501 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa
     4561 gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg cttacaattt
     4621 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgcggatc
     4681 tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg taccttctga
     4741 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4861 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4921 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4981 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     5041 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
     5101 gattcttctg acacaacagt ctcgaactta aggctagagc caccatgatt gaacaagatg
     5161 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5221 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5281 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5341 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5401 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5461 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5521 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5581 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5641 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5701 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5761 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5821 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5881 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5941 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgatggcc
     6001 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     6061 cgataaggat cctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat
     6121 tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtaatggtt tcttagacgt
     6181 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6241 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6301 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6361 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6421 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6481 gttttcgccc cgaagaacgt tttccaatga tgagcacttt caaagttctg ctatgtggcg
     6541 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6601 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6661 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6721 tgacaactat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6781 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6841 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6901 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6961 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7021 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7081 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7141 agataggtgc ctcactgatt aagcattggt aattcgaaat gaccgaccaa gcgacgccca
     7201 accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7261 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7321 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7381 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7441 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7501 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7561 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7621 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7681 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7741 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7801 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7861 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga
     7921 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7981 gattttggtc atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa
     8041 tacaggaacg cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc
     8101 ttcagtggat taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac
     8161 ggttaaaatt tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac
     8221 ctgtctgctg ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat
     8281 ggtaaatcca cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca
     8341 cgatgcgtcc ggcgtagagg atcgagatct
//
