LOCUS       sequence-423343-        8423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length humanBOP1. C-terminal HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8423)
  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 8423)
  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_BOP1:1-746
FEATURES             Location/Qualifiers
     source          1..8423
                     /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      1079..1088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3377..3397
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             3413..4297
                     /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(4369..4387)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4424..4545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4435..4454)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4489..4508
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4790..5147
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4790..4981
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4813..4833)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4998..5133
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5198..5992
                     /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(5252..5271)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5862..5881
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6056..6104
                     /note="synthetic polyadenylation signal"
     promoter        6260..6364
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6365..6433,6434..7225)
                     /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(6583..6602)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7383..7971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7463..7482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8087..8370
                     /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 ccgaattcgc
     1081 caccatggcg ggttcgcggg gtgcggggcg cacggcggcg ccgagcgtgc ggccggagaa
     1141 gcggcggtct gagcccgaac tggagcctga gcccgagccg gagccccccc tcctctgcac
     1201 ctctcctctc agccacagca ccggcagcga ttctggcgtc tccgacagcg aggagagtgt
     1261 gttctcaggc ctggaagatt ccggcagtga cagcagtgag gatgatgacg aaggcgacga
     1321 ggagggagag gacggagccc ttgatgacga gggccacagt gggattaaaa agaccactga
     1381 ggagcaggtg caggccagca ctccttgccc gaggacagag atggcgagcg cccggattgg
     1441 ggatgagtat gcggaggaca gctctgatga ggaggacatc cggaacacgg tgggcaacgt
     1501 gcccttggag tggtacgatg acttccccca cgtgggctac gacctggatg gcaggcgcat
     1561 ctacaagccc ctgcggaccc gggatgagct ggaccagttc ctggacaaga tggacgatcc
     1621 tgactactgg cgcaccgtgc aggacccgat gacagggcgg gacctgagac tgacggatga
     1681 gcaggtggcc ctggtgcggc ggctgcagag tggccagttt ggggatgtgg gcttcaaccc
     1741 ctatgagccg gctgtcgact tcttcagcgg ggacgtcatg atccacccgg tgaccaaccg
     1801 cccggccgac aagcgcagct tcatcccctc cctggtggag aaggagaagg tctctcgcat
     1861 ggtgcacgcc atcaagatgg gctggatcca gcctcgccgg ccccgagacc ccacccccag
     1921 cttctatgac ctgtgggccc aggaggaccc caacgccgtg ctcgggcgcc acaagatgca
     1981 cgtacctgct cccaagctgg ccctgccagg ccacgccgag tcgtacaacc caccccctga
     2041 atacctgctc agcgaggagg agcgcttggc gtgggaacag caggagccag gcgagaggaa
     2101 gctgagcttt ttgccacgca agttcccgag cctgcgggcc gtgcctgcct acggacgctt
     2161 catccaggaa cgcttcgagc gctgccttga cctgtacctg tgcccacggc agcgcaagat
     2221 gagggtgaat gtagaccctg aggacctcat ccccaagctg cctcggccga gggacctgca
     2281 gcccttcccc acgtgccagg ccctggtcta caggggccac agtgaccttg tccggtgcct
     2341 cagtgtctct cctgggggcc agtggctggt ttcaggctct gacgacggct ccctgcggct
     2401 ctgggaggtg gccactgccc gctgtgtgag gactgttccc gtggggggcg tggtgaagag
     2461 tgtggcctgg aaccccagcc ccgctgtctg cctggtggct gcagccgtgg aggactcggt
     2521 gctgctgctg aacccagctc tgggggaccg gctggtggcg ggcagcacag atcagctgtt
     2581 gagcgccttc gtcccgcctg aggagccccc cttgcagccg gcccgctggc tggaggcctc
     2641 agaggaggag cgccaagtgg gcctgcggct gcgcatctgc cacgggaagc cagtgacgca
     2701 ggtgacctgg cacgggcgtg gggactacct ggccgtggtg ctggccaccc aaggccacac
     2761 ccaggtgctg attcaccagc tgagccgtcg ccgcagccag agtccgttcc gccgcagcca
     2821 cggacaggtg cagcgagtgg ccttccaccc tgcccggccc ttcctgttgg tggcgtccca
     2881 gcgcagcgtc cgcctctacc acctgctgcg ccaggagctc accaagaagc tgatgcccaa
     2941 ctgcaagtgg gtgtccagcc tggcggtgca ccctgcaggt gacaacgtca tctgtgggag
     3001 ctacgatagc aagctggtgt ggtttgacct ggatctttcc accaagccat acaggatgct
     3061 gagacaccac aagaaggctc tgcgggctgt ggccttccac ccgcggtacc cactctttgc
     3121 gtcaggctcg gacgacggca gtgtcatcgt ctgccatggc atggtgtaca atgaccttct
     3181 gcagaacccc ttgctggtgc ccgtcaaggt gctgaaggga cacgtgctga cccgagatct
     3241 gggagtgctg gacgtcatct tccaccccac ccagccgtgg gtcttctcct cgggggcaga
     3301 cgggactgtc cgcctcttca cccggatatc tagatttggg cccaattcct gcaggcgagc
     3361 tctcgagcca accactgagg atctgtactt tcagagcgat aacgatggat ccgaaatcgg
     3421 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     3481 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     3541 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     3601 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     3661 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     3721 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     3781 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     3841 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     3901 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     3961 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     4021 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     4081 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     4141 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     4201 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     4261 gatcgcgcgc tggctgtcta ctctggagat ttccggttaa tagaattggc atgcaagctg
     4321 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     4381 aactagcata accccttggg gcggccgctt cgagcagaca tgataagata cattgatgag
     4441 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4501 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     4561 attcatttta tgtttcaggt tcagggggag atgtgggagg tttttttaag caagtaaaac
     4621 ctctacaaat gtggtaaaat cgaattttaa caaaatatta acgcttacaa tttcctgatg
     4681 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag
     4741 caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa
     4801 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4861 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4921 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4981 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5041 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     5101 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttgattctt
     5161 ctgacacaac agtctcgaac ttaaggctag agccaccatg attgaacaag atggattgca
     5221 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5281 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5341 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5401 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5461 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5521 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5581 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5641 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5701 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5761 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5821 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5881 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5941 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6001 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     6061 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     6121 gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg
     6181 gggtcagcac cgtttctgcg gactggcttt ctacgtaatg gtttcttaga cgtcaggtgg
     6241 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6301 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6361 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6421 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6481 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6541 ccccgaagaa cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg gcgcggtatt
     6601 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6661 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6721 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6781 tatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6841 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6901 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6961 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7021 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7081 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7141 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7201 tgcctcactg attaagcatt ggtaattcga aatgaccgac caagcgacgc ccaaccggta
     7261 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     7321 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7381 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7441 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7501 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7561 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7621 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7681 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7741 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7801 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     7861 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7921 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca agaagatcct
     7981 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8041 gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg aaatacagga
     8101 acgcacgctg gatggccctt cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg
     8161 gattaagtgg gggtaatgtg gcctgtaccc tctggttgca taggtattca tacggttaaa
     8221 atttatcagg cgcgattgcg gcagtttttc gggtggtttg ttgccatttt tacctgtctg
     8281 ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca attaagggat tatggtaaat
     8341 ccacttactg tctgccctcg tagccatcga gataaaccgc agtactccgg ccacgatgcg
     8401 tccggcgtag aggatcgaga tct
//
