LOCUS       sequence-425950-        5961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CHAF1B. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5961)
  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 5961)
  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_CHAF1B:1-559
FEATURES             Location/Qualifiers
     source          1..5961
                     /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    3024..3145
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3035..3054)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3260..3293)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3303..3350
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3382..4176
                     /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(3436..3455)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4046..4065
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4334..4922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4414..4433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5048..5096
                     /note="synthetic polyadenylation signal"
     CDS             complement(5120..5518)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5515..5524
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5549..5906)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5563..5698
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5715..5906
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5863..5883
                     /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 tgaaagtcat
     1261 cacttgtgaa atagcctggc acaacaagga gcccgtgtac agcctggact tccagcatgg
     1321 gacggctggg aggatccaca gactggcgtc tgccggcgtg gacaccaatg tcaggatctg
     1381 gaaggtagaa aagggaccag atggaaaagc catcgtggaa tttttgtcca atcttgctcg
     1441 tcataccaaa gccgtcaatg ttgtgcgttt ttctccaact ggggaaattt tagcatcggg
     1501 aggagatgat gctgtcatcc tattgtggaa ggtgaatgat aacaaggagc cggagcagat
     1561 cgcttttcag gatgaggacg aggcccagct gaacaaggag aactggacgg ttgtgaagac
     1621 tctgcggggc cacttagaag atgtgtatga tatttgctgg gcaactgatg ggaatttaat
     1681 ggcttctgcc tctgtggata acacagccat catatgggat gtcagcaaag gacaaaagat
     1741 atcaattttt aatgaacata aaagttatgt ccaaggagta acctgggacc ctttgggtca
     1801 atatgttgct actctgagct gtgacagggt gctgcgagta tacagtatac agaagaagcg
     1861 tgtggctttc aatgtttcga agatgctgtc tggaataggg gctgaaggag aggcaagaag
     1921 ctaccggatg tttcacgacg acagcatgaa gtctttcttc cgtagactga gtttcactcc
     1981 cgacggatct ttgcttctca cgccagctgg atgtgtggaa tctggtgaaa atgtaatgaa
     2041 taccacttat gttttctcca ggaagaatct taaaaggccc atcgctcatc ttccatgtcc
     2101 tggaaaagcc actcttgctg ttcgctgctg tccggtctac tttgaactga ggccagtggt
     2161 ggaaacaggt gtggagctga tgagtctgcc ctaccgcctg gtgtttgctg tggcctcgga
     2221 ggattccgtg cttctgtatg acacccagca gtccttccct tttggttacg tgtctaatat
     2281 acattaccac accctcagtg acatttcatg gtccagcgat ggtgccttcc tggccatttc
     2341 ttccacggac ggttactgct catttgtgac atttgagaaa gatgaacttg gaattccttt
     2401 gaaagagaag ccagttttga acatgagaac tcctgataca gcaaagaaaa ccaagagtca
     2461 gacacatcga gggtcttcgc caggacccag accggtagag ggaacccctg ccagcagaac
     2521 ccaagacccc agcagccccg gcacgactcc ccctcaggcc agacaggccc cagccccaac
     2581 agtcatcagg gaccctccct ccatcactcc tgctgtcaaa agccccttgc cggggccttc
     2641 ggaggagaag accctgcagc ccagtagtca aaacacaaaa gcccacccat cccggagggt
     2701 cactctgaac acactgcaag cctggagcaa gacaacaccc cggagaataa acttaacacc
     2761 cttaaagacg gacactccac caagttctgt accaaccagt gtgatttcca ccccttctac
     2821 agaagaaatt cagtcagaga cgcctggaga cgctcagggc agtcccccag agctaaagcg
     2881 gcccagactc gatgaaaaca aaggaggcac ggaaagtctg gacccttaat ctagaggtca
     2941 agcttcgggg gtttaaacct gcaggcatgc aagctgattt ggctgctgcc accgctgagc
     3001 aaggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3061 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3121 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3181 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     3241 cttaattaag acttctaata taacttcgta tagcatacat tatacgaagt tatggttcca
     3301 ctgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     3361 actaaaccca ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc
     3421 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3481 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3541 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3601 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3661 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3721 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3781 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3841 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3901 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3961 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4021 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4081 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4141 atcgccttct atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg
     4201 cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4261 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4321 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4381 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4441 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4501 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4561 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4621 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4681 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4741 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4801 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4861 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc
     4921 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4981 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg
     5041 gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg
     5101 ccatcgtgat ggctagagtt tagccctccc acacataacc agagggcagc aattcacgaa
     5161 tcccaactgc cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca
     5221 gatcgagaag cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc
     5281 cgatcgcgac gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca
     5341 cgagttctgc acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc
     5401 cgtcgctaga gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga
     5461 ttgtagccgt tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg
     5521 tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc
     5581 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
     5641 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     5701 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     5761 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     5821 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     5881 tccacagctg gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta
     5941 tttcaggcca tggtgctgcg c
//
