LOCUS       sequence-423444-        5504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PAAF1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5504)
  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 5504)
  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_PAAF1:1-392
FEATURES             Location/Qualifiers
     source          1..5504
                     /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"
     regulatory      1241..1250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2567..2688
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2578..2597)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2803..2836)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2846..2893
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2925..3719
                     /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(2979..2998)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3589..3608
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3877..4465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3957..3976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4591..4639
                     /note="synthetic polyadenylation signal"
     CDS             complement(4663..5061)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5058..5067
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5092..5449)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5106..5241
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5258..5449
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5406..5426
                     /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 ccatggcggc gcctttgagg
     1261 attcagagcg actgggcgca agccctcagg aaggatgaag gggaggcctg gctgagctgt
     1321 catcccccag ggaaaccatc tttgtatggc agcctgactt gtcaaggaat tggcctagat
     1381 ggcatcccag aggttacagt ttcagaagga tttactgtga atgaaataaa caagaaaagc
     1441 attcatattt catgtccaaa ggaaaatgca tcttctaagt ttttggcacc atatactact
     1501 ttttccagaa ttcatacaaa gagtataaca tgcctggaca tttccagcag aggaggtctt
     1561 ggtgtgtctt ctagtactga cgggaccatg aaaatctggc aggcttccaa tggagaactc
     1621 aggagagtat tggaaggaca tgtgtttgat gtgaattgtt gcaggttttt cccatcaggc
     1681 cttgtggtcc tgagtggggg aatggatgcc cagctgaaga tatggtcagc tgaagatgct
     1741 agctgcgtgg tgaccttcaa aggtcacaaa ggaggtatcc tggatacagc catcgttgat
     1801 cgggggagga atgtggtgtc tgcttctcga gatgggacag cacgactttg ggattgtggg
     1861 cgctcagcct gcttgggagt ccttgcagat tgtggttctt ctatcaatgg agtggcggtg
     1921 ggtgctgctg acaactccat aaaccttggc tcccctgagc agatgcccag tgaacgggag
     1981 gttggaacag aggccaaaat gctgctcttg gcccgggaag ataagaaact tcagtgcttg
     2041 ggactacaga gcaggcagct ggtgttcctc tttattggct cagacgcttt caactgctgt
     2101 acttttctct ctggcttctt gctattggct gggactcaag atggaaacat ttatcagctg
     2161 gatgtgagga gtccaagggc tccggtacaa gtcatccaca gatcaggagc accagttcta
     2221 tccctgctaa gtgtcagaga tggattcatt gctagccaag gtgatggaag ctgttttatt
     2281 gtccagcaag acttagacta tgtcactgag ctcactgggg ctgactgtga ccctgtgtac
     2341 aaggtagcca catgggagaa gcagatctac acatgctgtc gagacggtct tgtacgacgc
     2401 taccagcttt ctgacctcgg ctcgagcggt gggagctccg gtgtgagcgg ctggcggctg
     2461 ttcaagaaga ttagctaagt ttaaacggcc gcgactctag actgcaggca tgcaagctga
     2521 tttggctgct gccaccgctg agcaaggccg cttcgagcag acatgataag atacattgat
     2581 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2641 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2701 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa
     2761 aacctctaca aatgtggtaa aatcttaatt aagacttcta atataacttc gtatagcata
     2821 cattatacga agttatggtt ccactgttga caattaatca tcggcatagt atatcggcat
     2881 agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt gaacaagatg
     2941 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3001 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3061 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3121 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3181 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3241 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3301 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3361 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3421 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3481 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3541 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3601 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3661 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgat
     3721 tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg
     3781 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3841 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3901 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3961 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4021 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4081 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4141 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4201 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4261 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4321 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4381 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4441 tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4501 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4561 cacctagatc cttttatagt ccggaatcgt cacacaaaaa accaacacac agatgtaatg
     4621 aaaataaaga tattttattg cggccatcgt gatggctaga gtttagccct cccacacata
     4681 accagagggc agcaattcac gaatcccaac tgccgtcggc tgtccatcac tgtccttcac
     4741 tatggctttg atcccaggat gcagatcgag aagcacctgt cggcaccgtc cgcaggggct
     4801 caagatgccc ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc
     4861 agcagcagca gtgcccagca ccacgagttc tgcacaaggt cccccagtaa aatgatatac
     4921 attgacacca gtgaagatgc ggccgtcgct agagagagct gcgctggcga cgctgtagtc
     4981 ttcagagatg gggatgctgt tgattgtagc cgttgctctt tcaatgaggg tggattcttc
     5041 ttgagacaaa ggcttggcca tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa
     5101 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     5161 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     5221 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     5281 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     5341 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     5401 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga aggtacctaa
     5461 ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgc
//
