LOCUS       sequence-423377-        5898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human RBBP5. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5898)
  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 5898)
  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_RBBP5:1-538
FEATURES             Location/Qualifiers
     source          1..5898
                     /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    2961..3082
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2972..2991)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3197..3230)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3240..3287
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3319..4113
                     /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(3373..3392)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3983..4002
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4271..4859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4351..4370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4985..5033
                     /note="synthetic polyadenylation signal"
     CDS             complement(5057..5455)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5452..5461
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5486..5843)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5500..5635
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5652..5843
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5800..5820
                     /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 tgaacctcga
     1261 gttgctggag tcctttgggc agaactatcc agaggaagct gatggaactt tggattgtat
     1321 cagcatggct ttgacttgca cctttaacag gtggggcaca ctgcttgcag ttggctgtaa
     1381 tgatggccga attgtcatct gggatttctt gacaagaggc attgctaaaa taattagtgc
     1441 acacatccat ccagtgtgtt ctttatgctg gagccgagat ggtcataaac tcgtgagtgc
     1501 ttccactgat aacatagtgt cacagtggga tgttctttca ggcgactgtg accagaggtt
     1561 tcgattccct tcacccatct taaaagtcca atatcatcca cgagatcaga acaaggttct
     1621 cgtgtgtccc atgaaatctg ctcctgtcat gttgaccctt tcagattcca aacatgttgt
     1681 tctgccagtg gacgatgact ccgatttgaa cgttgtggca tcttttgata ggcgagggga
     1741 atatatttat acgggaaacg caaaaggcaa gattttggtc ctaaaaacag attctcagga
     1801 tcttgttgct tccttcagag tgacaactgg aacaagcaat accacagcca ttaagtcaat
     1861 tgagtttgcc cggaagggga gttgcttttt aattaacacg gcagatcgaa taatcagagt
     1921 ttatgatggc agagaaatct taacatgtgg aagagatgga gagcctgaac ctatgcagaa
     1981 attgcaggat ttggtgaata ggaccccatg gaagaaatgt tgtttctctg gggatgggga
     2041 atacatcgtg gcaggttctg cccggcagca tgccctgtac atctgggaga agagcattgg
     2101 caacctggtg aagattctcc atgggacgag aggagaactc ctcttggatg tagcttggca
     2161 tcctgttcga cccatcatag catccatttc cagtggagtg gtatctatct gggcacagaa
     2221 tcaagtagaa aactggagtg catttgcacc agacttcaaa gaattggatg aaaatgtaga
     2281 atacgaagaa agggaatcag agtttgatat tgaagatgaa gataagagtg agcctgagca
     2341 gacaggggct gatgctgcag aagatgagga agtggatgtc accagcgtgg accctattgc
     2401 tgccttctgt agcagtgatg aagagctgga agattcaaag gctctattgt atttacccat
     2461 tgcccctgag gtagaagacc cagaagaaaa tccttacggc cccccaccgg atgcagtcca
     2521 aacctccttg atggatgaag gggctagttc agagaagaag aggcagtcct cagcagatgg
     2581 gtcccagcca cctaagaaga aacccaaaac aaccaatata gaacttcaag gagtaccaaa
     2641 tgatgaagtc catccactac tgggtgtgaa gggggatggc aaatccaaga agaagcaagc
     2701 aggccggcct aaaggatcaa aaggtaaaga gaaagattct ccatttaaac cgaaactcta
     2761 caaaggggac agaggtttac ctctggaagg atcagcgaag ggtaaagtgc aggcggaact
     2821 cagccagccc ttgacagcag gaggagcaat ctcagaactg ttataatcta gaggtcaagc
     2881 ttcgggggtt taaacctgca ggcatgcaag ctgatttggc tgctgccacc gctgagcaag
     2941 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3001 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3061 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3121 gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatctt
     3181 aattaagact tctaatataa cttcgtatag catacattat acgaagttat ggttccactg
     3241 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     3301 aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3361 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3421 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3481 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3541 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3601 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3661 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3721 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3781 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3841 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3901 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3961 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4021 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4081 gccttctatc gccttcttga cgagttcttc tgattcgaaa tgaccgacca agcgacgccc
     4141 aaccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4201 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4261 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4321 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4381 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4441 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4501 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4561 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4621 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4681 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4741 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4801 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag
     4861 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4921 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa
     4981 tcgtcacaca aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca
     5041 tcgtgatggc tagagtttag ccctcccaca cataaccaga gggcagcaat tcacgaatcc
     5101 caactgccgt cggctgtcca tcactgtcct tcactatggc tttgatccca ggatgcagat
     5161 cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat gcccctgttc tcatttccga
     5221 tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc agcagtgccc agcaccacga
     5281 gttctgcaca aggtccccca gtaaaatgat atacattgac accagtgaag atgcggccgt
     5341 cgctagagag agctgcgctg gcgacgctgt agtcttcaga gatggggatg ctgttgattg
     5401 tagccgttgc tctttcaatg agggtggatt cttcttgaga caaaggcttg gccatggtgg
     5461 cgctagtgtc agaagaatcg agctttttgc aaaagcctag gcctccaaaa aagcctcctc
     5521 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     5581 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     5641 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
     5701 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
     5761 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
     5821 acagctggtt ctttccgcct cagaaggtac ctaaccaagt tcctctttca gaggttattt
     5881 caggccatgg tgctgcgc
//
