LOCUS       sequence-425884-        6005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CHAF1B. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6005)
  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 6005)
  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-C_CHAF1B:1-559
FEATURES             Location/Qualifiers
     source          1..6005
                     /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    3068..3189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3079..3098)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3304..3337)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3347..3394
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3426..4220
                     /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(3480..3499)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4090..4109
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4378..4966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4458..4477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5092..5140
                     /note="synthetic polyadenylation signal"
     CDS             complement(5164..5562)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5559..5568
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5593..5950)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5607..5742
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5759..5950
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5907..5927
                     /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 ataggggcga tcgcaacaag cttcgggaag
     1201 atctggagct agcggcgggg gatccggtgg gaattcgaca ccatgaaagt catcacttgt
     1261 gaaatagcct ggcacaacaa ggagcccgtg tacagcctgg acttccagca tgggacggct
     1321 gggaggatcc acagactggc gtctgccggc gtggacacca atgtcaggat ctggaaggta
     1381 gaaaagggac cagatggaaa agccatcgtg gaatttttgt ccaatcttgc tcgtcatacc
     1441 aaagccgtca atgttgtgcg tttttctcca actggggaaa ttttagcatc gggaggagat
     1501 gatgctgtca tcctattgtg gaaggtgaat gataacaagg agccggagca gatcgctttt
     1561 caggatgagg acgaggccca gctgaacaag gagaactgga cggttgtgaa gactctgcgg
     1621 ggccacttag aagatgtgta tgatatttgc tgggcaactg atgggaattt aatggcttct
     1681 gcctctgtgg ataacacagc catcatatgg gatgtcagca aaggacaaaa gatatcaatt
     1741 tttaatgaac ataaaagtta tgtccaagga gtaacctggg accctttggg tcaatatgtt
     1801 gctactctga gctgtgacag ggtgctgcga gtatacagta tacagaagaa gcgtgtggct
     1861 ttcaatgttt cgaagatgct gtctggaata ggggctgaag gagaggcaag aagctaccgg
     1921 atgtttcacg acgacagcat gaagtctttc ttccgtagac tgagtttcac tcccgacgga
     1981 tctttgcttc tcacgccagc tggatgtgtg gaatctggtg aaaatgtaat gaataccact
     2041 tatgttttct ccaggaagaa tcttaaaagg cccatcgctc atcttccatg tcctggaaaa
     2101 gccactcttg ctgttcgctg ctgtccggtc tactttgaac tgaggccagt ggtggaaaca
     2161 ggtgtggagc tgatgagtct gccctaccgc ctggtgtttg ctgtggcctc ggaggattcc
     2221 gtgcttctgt atgacaccca gcagtccttc ccttttggtt acgtgtctaa tatacattac
     2281 cacaccctca gtgacatttc atggtccagc gatggtgcct tcctggccat ttcttccacg
     2341 gacggttact gctcatttgt gacatttgag aaagatgaac ttggaattcc tttgaaagag
     2401 aagccagttt tgaacatgag aactcctgat acagcaaaga aaaccaagag tcagacacat
     2461 cgagggtctt cgccaggacc cagaccggta gagggaaccc ctgccagcag aacccaagac
     2521 cccagcagcc ccggcacgac tccccctcag gccagacagg ccccagcccc aacagtcatc
     2581 agggaccctc cctccatcac tcctgctgtc aaaagcccct tgccggggcc ttcggaggag
     2641 aagaccctgc agcccagtag tcaaaacaca aaagcccacc catcccggag ggtcactctg
     2701 aacacactgc aagcctggag caagacaaca ccccggagaa taaacttaac acccttaaag
     2761 acggacactc caccaagttc tgtaccaacc agtgtgattt ccaccccttc tacagaagaa
     2821 attcagtcag agacgcctgg agacgctcag ggcagtcccc cagagctaaa gcggcccaga
     2881 ctcgatgaaa acaaaggagg cacggaaagt ctggaccctg gctcgagcgg tgggagctcc
     2941 ggtgtgagcg gctggcggct gttcaagaag attagctaag tttaaacggc cgcgactcta
     3001 gactgcaggc atgcaagctg atttggctgc tgccaccgct gagcaaggcc gcttcgagca
     3061 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3121 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3181 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     3241 gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcttaat taagacttct
     3301 aatataactt cgtatagcat acattatacg aagttatggt tccactgttg acaattaatc
     3361 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa cccaggaggc
     3421 agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3481 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3541 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3601 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3661 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3721 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3781 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3841 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3901 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     3961 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4021 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4081 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4141 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4201 ttcttgacga gttcttctga ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc
     4261 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4321 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4381 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4441 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4501 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4561 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4621 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4681 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4741 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4801 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4861 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4921 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat
     4981 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5041 gagattatca aaaaggatct tcacctagat ccttttatag tccggaatcg tcacacaaaa
     5101 aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg tgatggctag
     5161 agtttagccc tcccacacat aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg
     5221 ctgtccatca ctgtccttca ctatggcttt gatcccagga tgcagatcga gaagcacctg
     5281 tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca tttccgatcg cgacgataca
     5341 agtcaggttg ccagctgccg cagcagcagc agtgcccagc accacgagtt ctgcacaagg
     5401 tcccccagta aaatgatata cattgacacc agtgaagatg cggccgtcgc tagagagagc
     5461 tgcgctggcg acgctgtagt cttcagagat ggggatgctg ttgattgtag ccgttgctct
     5521 ttcaatgagg gtggattctt cttgagacaa aggcttggcc atggtggcgc tagtgtcaga
     5581 agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     5641 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     5701 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     5761 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     5821 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     5881 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
     5941 tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag gccatggtgc
     6001 tgcgc
//
