LOCUS       sequence-425964-        5850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PRPF4. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5850)
  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 5850)
  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_PRPF4:1-522
FEATURES             Location/Qualifiers
     source          1..5850
                     /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    2913..3034
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2924..2943)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3149..3182)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3192..3239
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3271..4065
                     /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(3325..3344)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3935..3954
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4223..4811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4303..4322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4937..4985
                     /note="synthetic polyadenylation signal"
     CDS             complement(5009..5407)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5404..5413
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5438..5795)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5452..5587
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5604..5795
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5752..5772
                     /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 tggcttcctc
     1261 gcgagcctct tccacgcagg caaccaaaac taaagcaccc gacgacttag ttgctccggt
     1321 cgtgaagaaa ccacacatct attatggaag tttggaagag aaggagaggg agcgtctggc
     1381 caaaggagag tctgggattt tggggaaaga cggacttaaa gcagggatcg aagctggaaa
     1441 tattaatata acctctggag aagtgtttga aattgaagag catatcagcg agcgacaggc
     1501 agaagtattg gctgagtttg agagaaggaa gcgagcccgg cagatcaatg tttccacaga
     1561 tgactcagag gtcaaagctt gccttagagc cttgggggaa cccatcacac tttttggaga
     1621 gggtcctgct gaaagaagag aaaggttaag aaatatcctc tcagttgtcg gtactgatgc
     1681 cttgaaaaag accaaaaagg atgatgagaa gtctaaaaag tccaaagaag agtatcagca
     1741 aacctggtat catgaaggac caaatagctt gaaggtggca agactatgga ttgctaatta
     1801 ttcgttgccc agggcaatga aacgcttgga agaggcccga ctccataagg agattcctga
     1861 gacaacaagg acctcccaga tgcaagagct gcacaagtct ctccggtctt tgaataattt
     1921 ttgcagtcag attggggatg atcggcctat ctcctactgt cactttagtc ccaattccaa
     1981 gatgctggcc acagcttgtt ggagtgggct ttgcaagctc tggtctgttc ctgattgcaa
     2041 cctccttcac actcttcgag ggcataacac aaatgtagga gcaattgtat tccatcccaa
     2101 atccactgtc tccttggacc caaaagatgt caacctggcc tcttgtgcgg ctgatggctc
     2161 tgtgaagctt tggagtctcg acagtgatga accagtggca gatattgaag gccatacagt
     2221 gcgtgtggcg cgggtaatgt ggcatccttc aggacgtttc ctgggcacca cctgctatga
     2281 ccgttcatgg cgcttatggg atttggaggc tcaagaggag atcctgcatc aggaaggcca
     2341 tagcatgggt gtgtatgaca ttgccttcca tcaagatggc tctttggctg gcactggggg
     2401 actggatgca tttggtcgag tttgggacct acgcacagga cgttgtatca tgttcttaga
     2461 aggccacctg aaagaaatct atggaataaa tttctccccc aatggctatc acattgcaac
     2521 cggcagtggt gacaacacct gcaaagtgtg ggacctccga cagcggcgtt gcgtctacac
     2581 catccctgct catcagaact tagtgactgg tgtcaagttt gagcctatcc atgggaactt
     2641 cttgcttact ggtgcctatg ataacacagc caagatctgg acgcacccag gctggtcccc
     2701 gctgaagact ctggctggcc acgaaggcaa agtgatgggc ctagatattt cttccgatgg
     2761 gcagctcata gccacttgct catatgacag gaccttcaag ctgtggatgg ctgaataatc
     2821 tagaggtcaa gcttcggggg tttaaacctg caggcatgca agctgatttg gctgctgcca
     2881 ccgctgagca aggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     2941 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3001 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3061 gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg
     3121 tggtaaaatc ttaattaaga cttctaatat aacttcgtat agcatacatt atacgaagtt
     3181 atggttccac tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     3241 aggtgaggaa ctaaacccag gaggcagatc atgattgaac aagatggatt gcacgcaggt
     3301 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3361 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3421 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3481 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3541 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3601 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3661 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3721 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3781 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3841 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3901 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3961 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4021 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgattcga aatgaccgac
     4081 caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4141 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4201 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4261 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4321 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4381 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4441 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4501 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4561 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4621 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4681 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4741 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4801 aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4861 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4921 tatagtccgg aatcgtcaca caaaaaacca acacacagat gtaatgaaaa taaagatatt
     4981 ttattgcggc catcgtgatg gctagagttt agccctccca cacataacca gagggcagca
     5041 attcacgaat cccaactgcc gtcggctgtc catcactgtc cttcactatg gctttgatcc
     5101 caggatgcag atcgagaagc acctgtcggc accgtccgca ggggctcaag atgcccctgt
     5161 tctcatttcc gatcgcgacg atacaagtca ggttgccagc tgccgcagca gcagcagtgc
     5221 ccagcaccac gagttctgca caaggtcccc cagtaaaatg atatacattg acaccagtga
     5281 agatgcggcc gtcgctagag agagctgcgc tggcgacgct gtagtcttca gagatgggga
     5341 tgctgttgat tgtagccgtt gctctttcaa tgagggtgga ttcttcttga gacaaaggct
     5401 tggccatggt ggcgctagtg tcagaagaat cgagcttttt gcaaaagcct aggcctccaa
     5461 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     5521 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     5581 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     5641 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
     5701 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
     5761 gacacacatt ccacagctgg ttctttccgc ctcagaaggt acctaaccaa gttcctcttt
     5821 cagaggttat ttcaggccat ggtgctgcgc
//
