LOCUS       sequence-425940-        5709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human RRP9. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5709)
  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 5709)
  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_RRP9:1-475
FEATURES             Location/Qualifiers
     source          1..5709
                     /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    2772..2893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2783..2802)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3008..3041)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3051..3098
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3130..3924
                     /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(3184..3203)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3794..3813
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4082..4670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4162..4181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4796..4844
                     /note="synthetic polyadenylation signal"
     CDS             complement(4868..5266)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5263..5272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5297..5654)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5311..5446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5463..5654
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5611..5631
                     /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 tgtcggcaac
     1261 agcggctgct cgtaagcggg gaaagccggc ctctggggcc ggggctggcg cgggggccgg
     1321 caagcggcgg cgaaaggccg actctgcggg ggacaggggc aaatccaagg gtggcggcaa
     1381 gatgaatgag gagatctcca gcgactctga gagcgagagc ctagctccaa ggaagcctga
     1441 ggaggaggag gaggaggagc tggaggaaac tgcacaggaa aagaagctgc gcttggccaa
     1501 gctctaccta gagcagctcc gtcagcaaga ggaggagaag gctgaggccc gtgcatttga
     1561 ggaggaccag gtggcggggc gcctgaagga ggatgtgctt gagcagaggg gcaggctgca
     1621 gaagttggtg gcaaaagaga tccaggcccc agcctcagct gacattcgcg ttttacgggg
     1681 gcaccagctc tctatcacat gtttggtcgt cacccccgat gactcagcca tcttctctgc
     1741 tgccaaagac tgcagcatca ttaagtggag cgtggagagt ggacggaagc tgcatgtgat
     1801 tcctcgagcc aagaagggtg ccgagggaaa gccccctggc cacagcagcc acgtcctctg
     1861 catggccatc tcctccgacg gcaagtacct tgcctctggt gaccgcagca agctcattct
     1921 catttgggag gcccagagct gccagcactt gtacaccttc acaggacacc gggatgcagt
     1981 gtcgggtctg gcattccgca gaggcaccca ccagctctac agcacatccc acgatcgctc
     2041 cgtgaaggtg tggaatgtgg cagagaactc ctacgtggag acgctcttcg gacaccagga
     2101 cgctgtggct gcactggatg ccttgagccg ggagtgctgt gtgacggctg ggggccggga
     2161 tgggactgta cgtgtgtgga agatccccga ggagtcccag cttgtcttct atggccacca
     2221 gggctccatc gactgcatcc acctaatcaa tgaggagcac atggtgtccg gcgcggacga
     2281 tggctctgtg gccttgtggg gtctctccaa gaagcgacca cttgccctgc agcgtgaagc
     2341 tcacgggctg cggggagagc caggcctgga gcagcccttc tggatatcgt cggtggcagc
     2401 cctcctcaac acagaccttg tggccacagg ctcccacagc tcctgtgtgc ggctttggca
     2461 gtgtggggaa ggcttccggc agcttgacct tctctgtgac atccccctgg tgggttttat
     2521 caacagcctc aagttctcca gctctgggga cttcctggtg gctggggtag ggcaggagca
     2581 caggcttggc cgatggtgga gaatcaaaga ggctcggaat tctgtctgca tcatcccact
     2641 ccgcagggtc cctgtacccc cagctgctgg ttcctaatct agaggtcaag cttcgggggt
     2701 ttaaacctgc aggcatgcaa gctgatttgg ctgctgccac cgctgagcaa ggccgcttcg
     2761 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2821 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2881 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     2941 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatct taattaagac
     3001 ttctaatata acttcgtata gcatacatta tacgaagtta tggttccact gttgacaatt
     3061 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg
     3121 aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3181 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3241 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3301 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3361 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3421 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3481 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3541 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3601 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3661 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3721 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3781 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3841 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3901 cgccttcttg acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat
     3961 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4021 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4081 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4141 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4201 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4261 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4321 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4381 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4441 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4501 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4561 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4621 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt
     4681 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4741 tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga atcgtcacac
     4801 aaaaaaccaa cacacagatg taatgaaaat aaagatattt tattgcggcc atcgtgatgg
     4861 ctagagttta gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg
     4921 tcggctgtcc atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca
     4981 cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga
     5041 tacaagtcag gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac
     5101 aaggtccccc agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga
     5161 gagctgcgct ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg
     5221 ctctttcaat gagggtggat tcttcttgag acaaaggctt ggccatggtg gcgctagtgt
     5281 cagaagaatc gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct
     5341 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc
     5401 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
     5461 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     5521 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc
     5581 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagctggt
     5641 tctttccgcc tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg
     5701 gtgctgcgc
//
