LOCUS       sequence-425985-        6099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human BTRC. a N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6099)
  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 6099)
  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: pBiT3.1-N_BTRC:1-605
FEATURES             Location/Qualifiers
     source          1..6099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3162..3283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3173..3192)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3398..3431)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3441..3488
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3520..4314
                     /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(3574..3593)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4184..4203
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4472..5060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4552..4571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5186..5234
                     /note="synthetic polyadenylation signal"
     CDS             complement(5258..5656)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5653..5662
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5687..6044)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5701..5836
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5853..6044
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6001..6021
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggtta gcgatcgcca tggtgagcgg
     1201 ctggcggctg ttcaagaaga ttagcgggag ctccggtggc tcgagcggta tggacccggc
     1261 cgaggcggtg ctgcaagaga aggcactcaa gtttatgtgc tctatgccca ggtctctgtg
     1321 gctgggctgc tccagcctgg cggacagcat gccttcgctg cgatgcctgt ataacccagg
     1381 gactggcgca ctcacagctt tccagaattc ctcagagaga gaagactgta ataatggcga
     1441 accccctagg aagataatac cagagaagaa ttcacttaga cagacataca acagctgtgc
     1501 cagactctgc ttaaaccaag aaacagtatg tttagcaagc actgctatga agactgagaa
     1561 ttgtgtggcc aaaacaaaac ttgccaatgg cacttccagt atgattgtgc ccaagcaacg
     1621 gaaactctca gcaagctatg aaaaggaaaa ggaactgtgt gtcaaatact ttgagcagtg
     1681 gtcagagtca gatcaagtgg aatttgtgga acatcttata tcccaaatgt gtcattacca
     1741 acatgggcac ataaactcgt atcttaaacc tatgttgcag agagatttca taactgctct
     1801 gccagctcgg ggattggatc atattgctga gaacattctg tcatacctgg atgccaaatc
     1861 actatgtgct gctgaacttg tgtgcaagga atggtaccga gtgacctctg atggcatgct
     1921 gtggaagaag cttatcgaga gaatggtcag gacagattct ctgtggagag gcctggcaga
     1981 acgaagagga tggggacagt atttattcaa aaacaaacct cctgacggga atgctcctcc
     2041 caactctttt tatagagcac tttatcctaa aattatacaa gacattgaga caatagaatc
     2101 taattggaga tgtggaagac atagtttaca gagaattcac tgccgaagtg aaacaagcaa
     2161 aggagtttac tgtttacagt atgatgatca gaaaatagta agcggccttc gagacaacac
     2221 aatcaagatc tgggataaaa acacattgga atgcaagcga attctcacag gccatacagg
     2281 ttcagtcctc tgtctccagt atgatgagag agtgatcata acaggatcat cggattccac
     2341 ggtcagagtg tgggatgtaa atacaggtga aatgctaaac acgttgattc accattgtga
     2401 agcagttctg cacttgcgtt tcaataatgg catgatggtg acctgctcca aagatcgttc
     2461 cattgctgta tgggatatgg cctccccaac tgacattacc ctccggaggg tgctggtcgg
     2521 acaccgagct gctgtcaatg ttgtagactt tgatgacaag tacattgttt ctgcatctgg
     2581 ggatagaact ataaaggtat ggaacacaag tacttgtgaa tttgtaagga ccttaaatgg
     2641 acacaaacga ggcattgcct gtttgcagta cagggacagg ctggtagtga gtggctcatc
     2701 tgacaacact atcagattat gggacataga atgtggtgca tgtttacgag tgttagaagg
     2761 ccatgaggaa ttggtgcgtt gtattcgatt tgataacaag aggatagtca gtggggccta
     2821 tgatggaaaa attaaagtgt gggatcttgt ggctgctttg gacccccgtg ctcctgcagg
     2881 gacactctgt ctacggaccc ttgtggagca ttccggaaga gtttttcgac tacagtttga
     2941 tgaattccag attgtcagta gttcacatga tgacacaatc ctcatctggg acttcctaaa
     3001 tgatccagct gcccaagctg aacccccccg ttccccttct cgaacataca cctacatctc
     3061 cagataatct agaggtcaag cttcgggggt ttaaacctgc aggcatgcaa gctgatttgg
     3121 ctgctgccac cgctgagcaa ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3181 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3241 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3301 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     3361 ctacaaatgt ggtaaaatct taattaagac ttctaatata acttcgtata gcatacatta
     3421 tacgaagtta tggttccact gttgacaatt aatcatcggc atagtatatc ggcatagtat
     3481 aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg
     3541 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3601 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3661 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3721 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3781 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3841 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3901 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3961 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4021 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4081 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4141 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4201 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4261 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa
     4321 atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat
     4381 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4441 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4501 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4561 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4621 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4681 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4741 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4801 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4861 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4921 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4981 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5041 gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5101 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5161 agatcctttt atagtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat
     5221 aaagatattt tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag
     5281 agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg
     5341 ctttgatccc aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga
     5401 tgcccctgtt ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag
     5461 cagcagtgcc cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga
     5521 caccagtgaa gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag
     5581 agatggggat gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag
     5641 acaaaggctt ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta
     5701 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     5761 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     5821 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     5881 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     5941 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     6001 accctaactg acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag
     6061 ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
