LOCUS       sequence-425916-        5460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PAAF1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5460)
  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 5460)
  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_PAAF1:1-392
FEATURES             Location/Qualifiers
     source          1..5460
                     /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    2523..2644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2534..2553)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2759..2792)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2802..2849
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2881..3675
                     /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(2935..2954)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3545..3564
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3833..4421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3913..3932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4547..4595
                     /note="synthetic polyadenylation signal"
     CDS             complement(4619..5017)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5014..5023
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5048..5405)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5062..5197
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5214..5405
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5362..5382
                     /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 tggcggcgcc
     1261 tttgaggatt cagagcgact gggcgcaagc cctcaggaag gatgaagggg aggcctggct
     1321 gagctgtcat cccccaggga aaccatcttt gtatggcagc ctgacttgtc aaggaattgg
     1381 cctagatggc atcccagagg ttacagtttc agaaggattt actgtgaatg aaataaacaa
     1441 gaaaagcatt catatttcat gtccaaagga aaatgcatct tctaagtttt tggcaccata
     1501 tactactttt tccagaattc atacaaagag tataacatgc ctggacattt ccagcagagg
     1561 aggtcttggt gtgtcttcta gtactgacgg gaccatgaaa atctggcagg cttccaatgg
     1621 agaactcagg agagtattgg aaggacatgt gtttgatgtg aattgttgca ggtttttccc
     1681 atcaggcctt gtggtcctga gtgggggaat ggatgcccag ctgaagatat ggtcagctga
     1741 agatgctagc tgcgtggtga ccttcaaagg tcacaaagga ggtatcctgg atacagccat
     1801 cgttgatcgg gggaggaatg tggtgtctgc ttctcgagat gggacagcac gactttggga
     1861 ttgtgggcgc tcagcctgct tgggagtcct tgcagattgt ggttcttcta tcaatggagt
     1921 ggcggtgggt gctgctgaca actccataaa ccttggctcc cctgagcaga tgcccagtga
     1981 acgggaggtt ggaacagagg ccaaaatgct gctcttggcc cgggaagata agaaacttca
     2041 gtgcttggga ctacagagca ggcagctggt gttcctcttt attggctcag acgctttcaa
     2101 ctgctgtact tttctctctg gcttcttgct attggctggg actcaagatg gaaacattta
     2161 tcagctggat gtgaggagtc caagggctcc ggtacaagtc atccacagat caggagcacc
     2221 agttctatcc ctgctaagtg tcagagatgg attcattgct agccaaggtg atggaagctg
     2281 ttttattgtc cagcaagact tagactatgt cactgagctc actggggctg actgtgaccc
     2341 tgtgtacaag gtagccacat gggagaagca gatctacaca tgctgtcgag acggtcttgt
     2401 acgacgctac cagctttctg acctctaatc tagaggtcaa gcttcggggg tttaaacctg
     2461 caggcatgca agctgatttg gctgctgcca ccgctgagca aggccgcttc gagcagacat
     2521 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2581 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2641 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     2701 ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc ttaattaaga cttctaatat
     2761 aacttcgtat agcatacatt atacgaagtt atggttccac tgttgacaat taatcatcgg
     2821 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc
     2881 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     2941 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3001 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3061 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3121 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3181 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3241 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3301 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3361 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3421 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3481 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3541 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3601 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3661 gacgagttct tctgattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact
     3721 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3781 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3841 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3901 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3961 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4021 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4081 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4141 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4201 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4261 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4321 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4381 tttgcaagca gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt
     4441 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4501 tatcaaaaag gatcttcacc tagatccttt tatagtccgg aatcgtcaca caaaaaacca
     4561 acacacagat gtaatgaaaa taaagatatt ttattgcggc catcgtgatg gctagagttt
     4621 agccctccca cacataacca gagggcagca attcacgaat cccaactgcc gtcggctgtc
     4681 catcactgtc cttcactatg gctttgatcc caggatgcag atcgagaagc acctgtcggc
     4741 accgtccgca ggggctcaag atgcccctgt tctcatttcc gatcgcgacg atacaagtca
     4801 ggttgccagc tgccgcagca gcagcagtgc ccagcaccac gagttctgca caaggtcccc
     4861 cagtaaaatg atatacattg acaccagtga agatgcggcc gtcgctagag agagctgcgc
     4921 tggcgacgct gtagtcttca gagatgggga tgctgttgat tgtagccgtt gctctttcaa
     4981 tgagggtgga ttcttcttga gacaaaggct tggccatggt ggcgctagtg tcagaagaat
     5041 cgagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     5101 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     5161 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     5221 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     5281 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     5341 gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc
     5401 ctcagaaggt acctaaccaa gttcctcttt cagaggttat ttcaggccat ggtgctgcgc
//
