LOCUS       sequence-425872-        5840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PRPF19. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5840)
  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 5840)
  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_PRPF19:1-504
FEATURES             Location/Qualifiers
     source          1..5840
                     /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    2903..3024
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2914..2933)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3139..3172)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3182..3229
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3261..4055
                     /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(3315..3334)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3925..3944
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4213..4801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4293..4312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4927..4975
                     /note="synthetic polyadenylation signal"
     CDS             complement(4999..5397)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5394..5403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5428..5785)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5442..5577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5594..5785
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5742..5762
                     /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 ccatgtccct aatctgctcc
     1261 atctctaacg aagtgccgga gcacccatgt gtatcccctg tctctaatca tgtttatgag
     1321 cggcggctca tcgagaagta cattgcggag aatggtaccg accccatcaa caaccagcct
     1381 ctctccgagg agcagctcat cgacatcaaa gttgctcacc caatccggcc caagcctccc
     1441 tcagccacca gcatcccggc cattctgaaa gctttgcagg atgagtggga tgcagtcatg
     1501 ctgcacagct tcactctgcg ccagcagctg cagacaaccc gccaagagct gtcacacgct
     1561 ctgtaccagc acgatgccgc ctgccgtgtc attgcccgtc tcaccaagga agtcactgct
     1621 gcccgagaag ctctggctac cctgaaacca caggctggcc tcattgtgcc ccaggctgtg
     1681 ccaagttccc aaccaagtgt tgtgggtgcg ggtgagccaa tggatttggg tgagctggtg
     1741 ggaatgaccc cagagattat tcagaagctt caagacaaag ccactgtgct aaccacggag
     1801 cgcaagaaga gagggaagac tgtgcctgag gagctggtga agccagaaga gctcagcaaa
     1861 taccggcagg tggcatccca cgtggggttg cacagtgcca gcattcctgg gatcctggcc
     1921 ctggacctct gcccgtccga caccaacaag atcctcactg gtggggcgga taaaaatgtc
     1981 gttgtgtttg acaaaagttc tgaacaaatc ctggctaccc tcaaaggcca taccaagaag
     2041 gtcaccagcg tggtgtttca cccttcccag gacctggtgt tttctgcttc ccccgatgcc
     2101 actatcagga tttggtcggt ccccaatgcc tcttgtgtac aggtggttcg ggcccatgag
     2161 agtgctgtga caggcctcag ccttcatgcc actggcgact atctcctgag ctcctccgat
     2221 gatcagtact gggctttctc tgacatccag acagggcgtg tgctcaccaa ggtgacagat
     2281 gagacctccg gctgctctct cacctgtgca cagttccacc ctgacggact catctttgga
     2341 acaggaacca tggactctca gatcaagatc tgggacttga aggaacgtac taatgtggcc
     2401 aacttccctg gccactcggg ccccatcact agcatcgcct tctctgagaa tggttactac
     2461 ctggctacag cggctgatga ctcctctgtc aagctctggg atctgcgcaa gcttaagaac
     2521 tttaagactt tgcagctgga taacaacttt gaggtaaagt cactgatctt tgaccagagt
     2581 ggtacctacc tggctcttgg gggcacggat gtccagatct acatctgcaa acaatggacg
     2641 gagattcttc actttacaga gcatagcggc ctgaccacag gggtggcctt cgggcatcac
     2701 gccaagttca tcgcttcaac aggcatggac agaagcctca agttctacag cctgggctcg
     2761 agcggtggga gctccggtgt gagcggctgg cggctgttca agaagattag ctaagtttaa
     2821 acggccgcga ctctagactg caggcatgca agctgatttg gctgctgcca ccgctgagca
     2881 aggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2941 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3001 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3061 cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc
     3121 ttaattaaga cttctaatat aacttcgtat agcatacatt atacgaagtt atggttccac
     3181 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     3241 ctaaacccag gaggcagatc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3301 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3361 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3421 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3481 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3541 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3601 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3661 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3721 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3781 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3841 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3901 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3961 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4021 tcgccttcta tcgccttctt gacgagttct tctgattcga aatgaccgac caagcgacgc
     4081 ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4141 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4201 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4261 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4321 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4381 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4441 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4501 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4561 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4621 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     4681 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4741 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca
     4801 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4861 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg
     4921 aatcgtcaca caaaaaacca acacacagat gtaatgaaaa taaagatatt ttattgcggc
     4981 catcgtgatg gctagagttt agccctccca cacataacca gagggcagca attcacgaat
     5041 cccaactgcc gtcggctgtc catcactgtc cttcactatg gctttgatcc caggatgcag
     5101 atcgagaagc acctgtcggc accgtccgca ggggctcaag atgcccctgt tctcatttcc
     5161 gatcgcgacg atacaagtca ggttgccagc tgccgcagca gcagcagtgc ccagcaccac
     5221 gagttctgca caaggtcccc cagtaaaatg atatacattg acaccagtga agatgcggcc
     5281 gtcgctagag agagctgcgc tggcgacgct gtagtcttca gagatgggga tgctgttgat
     5341 tgtagccgtt gctctttcaa tgagggtgga ttcttcttga gacaaaggct tggccatggt
     5401 ggcgctagtg tcagaagaat cgagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     5461 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     5521 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     5581 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     5641 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     5701 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     5761 ccacagctgg ttctttccgc ctcagaaggt acctaaccaa gttcctcttt cagaggttat
     5821 ttcaggccat ggtgctgcgc
//
