LOCUS       sequence-425847-        5894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PRPF4. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5894)
  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 5894)
  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_PRPF4:1-522
FEATURES             Location/Qualifiers
     source          1..5894
                     /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    2957..3078
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2968..2987)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3193..3226)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3236..3283
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3315..4109
                     /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(3369..3388)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3979..3998
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4267..4855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4347..4366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4981..5029
                     /note="synthetic polyadenylation signal"
     CDS             complement(5053..5451)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5448..5457
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5482..5839)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5496..5631
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5648..5839
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5796..5816
                     /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 ccatggcttc ctcgcgagcc
     1261 tcttccacgc aggcaaccaa aactaaagca cccgacgact tagttgctcc ggtcgtgaag
     1321 aaaccacaca tctattatgg aagtttggaa gagaaggaga gggagcgtct ggccaaagga
     1381 gagtctggga ttttggggaa agacggactt aaagcaggga tcgaagctgg aaatattaat
     1441 ataacctctg gagaagtgtt tgaaattgaa gagcatatca gcgagcgaca ggcagaagta
     1501 ttggctgagt ttgagagaag gaagcgagcc cggcagatca atgtttccac agatgactca
     1561 gaggtcaaag cttgccttag agccttgggg gaacccatca cactttttgg agagggtcct
     1621 gctgaaagaa gagaaaggtt aagaaatatc ctctcagttg tcggtactga tgccttgaaa
     1681 aagaccaaaa aggatgatga gaagtctaaa aagtccaaag aagagtatca gcaaacctgg
     1741 tatcatgaag gaccaaatag cttgaaggtg gcaagactat ggattgctaa ttattcgttg
     1801 cccagggcaa tgaaacgctt ggaagaggcc cgactccata aggagattcc tgagacaaca
     1861 aggacctccc agatgcaaga gctgcacaag tctctccggt ctttgaataa tttttgcagt
     1921 cagattgggg atgatcggcc tatctcctac tgtcacttta gtcccaattc caagatgctg
     1981 gccacagctt gttggagtgg gctttgcaag ctctggtctg ttcctgattg caacctcctt
     2041 cacactcttc gagggcataa cacaaatgta ggagcaattg tattccatcc caaatccact
     2101 gtctccttgg acccaaaaga tgtcaacctg gcctcttgtg cggctgatgg ctctgtgaag
     2161 ctttggagtc tcgacagtga tgaaccagtg gcagatattg aaggccatac agtgcgtgtg
     2221 gcgcgggtaa tgtggcatcc ttcaggacgt ttcctgggca ccacctgcta tgaccgttca
     2281 tggcgcttat gggatttgga ggctcaagag gagatcctgc atcaggaagg ccatagcatg
     2341 ggtgtgtatg acattgcctt ccatcaagat ggctctttgg ctggcactgg gggactggat
     2401 gcatttggtc gagtttggga cctacgcaca ggacgttgta tcatgttctt agaaggccac
     2461 ctgaaagaaa tctatggaat aaatttctcc cccaatggct atcacattgc aaccggcagt
     2521 ggtgacaaca cctgcaaagt gtgggacctc cgacagcggc gttgcgtcta caccatccct
     2581 gctcatcaga acttagtgac tggtgtcaag tttgagccta tccatgggaa cttcttgctt
     2641 actggtgcct atgataacac agccaagatc tggacgcacc caggctggtc cccgctgaag
     2701 actctggctg gccacgaagg caaagtgatg ggcctagata tttcttccga tgggcagctc
     2761 atagccactt gctcatatga caggaccttc aagctgtgga tggctgaagg ctcgagcggt
     2821 gggagctccg gtgtgagcgg ctggcggctg ttcaagaaga ttagctaagt ttaaacggcc
     2881 gcgactctag actgcaggca tgcaagctga tttggctgct gccaccgctg agcaaggccg
     2941 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3001 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3061 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3121 gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcttaatt
     3181 aagacttcta atataacttc gtatagcata cattatacga agttatggtt ccactgttga
     3241 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     3301 ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3361 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3421 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3481 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3541 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3601 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3661 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3721 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3781 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3841 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3901 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     3961 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4021 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4081 tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc
     4141 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4201 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4261 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4321 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4381 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4441 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4501 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4561 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4621 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4681 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4741 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4801 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga
     4861 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4921 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt ccggaatcgt
     4981 cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg cggccatcgt
     5041 gatggctaga gtttagccct cccacacata accagagggc agcaattcac gaatcccaac
     5101 tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat gcagatcgag
     5161 aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat ttccgatcgc
     5221 gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca ccacgagttc
     5281 tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc ggccgtcgct
     5341 agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt tgattgtagc
     5401 cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca tggtggcgct
     5461 agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     5521 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     5581 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     5641 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5701 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     5761 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     5821 ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg ttatttcagg
     5881 ccatggtgct gcgc
//
