LOCUS       sequence-425962-        5796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PRPF19. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5796)
  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 5796)
  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_PRPF19:1-504
FEATURES             Location/Qualifiers
     source          1..5796
                     /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    2859..2980
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2870..2889)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3095..3128)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3138..3185
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3217..4011
                     /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(3271..3290)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3881..3900
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4169..4757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4249..4268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4883..4931
                     /note="synthetic polyadenylation signal"
     CDS             complement(4955..5353)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5350..5359
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5384..5741)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5398..5533
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5550..5741
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5698..5718
                     /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 tgtccctaat
     1261 ctgctccatc tctaacgaag tgccggagca cccatgtgta tcccctgtct ctaatcatgt
     1321 ttatgagcgg cggctcatcg agaagtacat tgcggagaat ggtaccgacc ccatcaacaa
     1381 ccagcctctc tccgaggagc agctcatcga catcaaagtt gctcacccaa tccggcccaa
     1441 gcctccctca gccaccagca tcccggccat tctgaaagct ttgcaggatg agtgggatgc
     1501 agtcatgctg cacagcttca ctctgcgcca gcagctgcag acaacccgcc aagagctgtc
     1561 acacgctctg taccagcacg atgccgcctg ccgtgtcatt gcccgtctca ccaaggaagt
     1621 cactgctgcc cgagaagctc tggctaccct gaaaccacag gctggcctca ttgtgcccca
     1681 ggctgtgcca agttcccaac caagtgttgt gggtgcgggt gagccaatgg atttgggtga
     1741 gctggtggga atgaccccag agattattca gaagcttcaa gacaaagcca ctgtgctaac
     1801 cacggagcgc aagaagagag ggaagactgt gcctgaggag ctggtgaagc cagaagagct
     1861 cagcaaatac cggcaggtgg catcccacgt ggggttgcac agtgccagca ttcctgggat
     1921 cctggccctg gacctctgcc cgtccgacac caacaagatc ctcactggtg gggcggataa
     1981 aaatgtcgtt gtgtttgaca aaagttctga acaaatcctg gctaccctca aaggccatac
     2041 caagaaggtc accagcgtgg tgtttcaccc ttcccaggac ctggtgtttt ctgcttcccc
     2101 cgatgccact atcaggattt ggtcggtccc caatgcctct tgtgtacagg tggttcgggc
     2161 ccatgagagt gctgtgacag gcctcagcct tcatgccact ggcgactatc tcctgagctc
     2221 ctccgatgat cagtactggg ctttctctga catccagaca gggcgtgtgc tcaccaaggt
     2281 gacagatgag acctccggct gctctctcac ctgtgcacag ttccaccctg acggactcat
     2341 ctttggaaca ggaaccatgg actctcagat caagatctgg gacttgaagg aacgtactaa
     2401 tgtggccaac ttccctggcc actcgggccc catcactagc atcgccttct ctgagaatgg
     2461 ttactacctg gctacagcgg ctgatgactc ctctgtcaag ctctgggatc tgcgcaagct
     2521 taagaacttt aagactttgc agctggataa caactttgag gtaaagtcac tgatctttga
     2581 ccagagtggt acctacctgg ctcttggggg cacggatgtc cagatctaca tctgcaaaca
     2641 atggacggag attcttcact ttacagagca tagcggcctg accacagggg tggccttcgg
     2701 gcatcacgcc aagttcatcg cttcaacagg catggacaga agcctcaagt tctacagcct
     2761 gtaatctaga ggtcaagctt cgggggttta aacctgcagg catgcaagct gatttggctg
     2821 ctgccaccgc tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg
     2881 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2941 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3001 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     3061 caaatgtggt aaaatcttaa ttaagacttc taatataact tcgtatagca tacattatac
     3121 gaagttatgg ttccactgtt gacaattaat catcggcata gtatatcggc atagtataat
     3181 acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac
     3241 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3301 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3361 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3421 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3481 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3541 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3601 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3661 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3721 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3781 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3841 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3901 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3961 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg
     4021 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     4081 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4141 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4201 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4261 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4321 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4381 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4441 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4501 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4561 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4621 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4681 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4741 gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4801 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4861 tccttttata gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa
     4921 gatattttat tgcggccatc gtgatggcta gagtttagcc ctcccacaca taaccagagg
     4981 gcagcaattc acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt
     5041 tgatcccagg atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc
     5101 ccctgttctc atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag
     5161 cagtgcccag caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac
     5221 cagtgaagat gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga
     5281 tggggatgct gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca
     5341 aaggcttggc catggtggcg ctagtgtcag aagaatcgag ctttttgcaa aagcctaggc
     5401 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     5461 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     5521 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     5581 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
     5641 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     5701 ctaactgaca cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc
     5761 ctctttcaga ggttatttca ggccatggtg ctgcgc
//
