LOCUS       sequence-423346-        7799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human RBBP5. C-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7799)
  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 7799)
  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: pHTC-HaloTag_RBBP5:1-538
FEATURES             Location/Qualifiers
     source          1..7799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2753..2773
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2789..3673
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(3745..3763)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3800..3921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3811..3830)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3865..3884
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4166..4523
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4166..4357
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4189..4209)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4374..4509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4574..5368
                     /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(4628..4647)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5238..5257
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5432..5480
                     /note="synthetic polyadenylation signal"
     promoter        5636..5740
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5741..5809,5810..6601)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5959..5978)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6759..7347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6839..6858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7463..7746
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcaaa gcgatcgctt ccgaattcgc
     1081 caccatgaac ctcgagttgc tggagtcctt tgggcagaac tatccagagg aagctgatgg
     1141 aactttggat tgtatcagca tggctttgac ttgcaccttt aacaggtggg gcacactgct
     1201 tgcagttggc tgtaatgatg gccgaattgt catctgggat ttcttgacaa gaggcattgc
     1261 taaaataatt agtgcacaca tccatccagt gtgttcttta tgctggagcc gagatggtca
     1321 taaactcgtg agtgcttcca ctgataacat agtgtcacag tgggatgttc tttcaggcga
     1381 ctgtgaccag aggtttcgat tcccttcacc catcttaaaa gtccaatatc atccacgaga
     1441 tcagaacaag gttctcgtgt gtcccatgaa atctgctcct gtcatgttga ccctttcaga
     1501 ttccaaacat gttgttctgc cagtggacga tgactccgat ttgaacgttg tggcatcttt
     1561 tgataggcga ggggaatata tttatacggg aaacgcaaaa ggcaagattt tggtcctaaa
     1621 aacagattct caggatcttg ttgcttcctt cagagtgaca actggaacaa gcaataccac
     1681 agccattaag tcaattgagt ttgcccggaa ggggagttgc tttttaatta acacggcaga
     1741 tcgaataatc agagtttatg atggcagaga aatcttaaca tgtggaagag atggagagcc
     1801 tgaacctatg cagaaattgc aggatttggt gaataggacc ccatggaaga aatgttgttt
     1861 ctctggggat ggggaataca tcgtggcagg ttctgcccgg cagcatgccc tgtacatctg
     1921 ggagaagagc attggcaacc tggtgaagat tctccatggg acgagaggag aactcctctt
     1981 ggatgtagct tggcatcctg ttcgacccat catagcatcc atttccagtg gagtggtatc
     2041 tatctgggca cagaatcaag tagaaaactg gagtgcattt gcaccagact tcaaagaatt
     2101 ggatgaaaat gtagaatacg aagaaaggga atcagagttt gatattgaag atgaagataa
     2161 gagtgagcct gagcagacag gggctgatgc tgcagaagat gaggaagtgg atgtcaccag
     2221 cgtggaccct attgctgcct tctgtagcag tgatgaagag ctggaagatt caaaggctct
     2281 attgtattta cccattgccc ctgaggtaga agacccagaa gaaaatcctt acggcccccc
     2341 accggatgca gtccaaacct ccttgatgga tgaaggggct agttcagaga agaagaggca
     2401 gtcctcagca gatgggtccc agccacctaa gaagaaaccc aaaacaacca atatagaact
     2461 tcaaggagta ccaaatgatg aagtccatcc actactgggt gtgaaggggg atggcaaatc
     2521 caagaagaag caagcaggcc ggcctaaagg atcaaaaggt aaagagaaag attctccatt
     2581 taaaccgaaa ctctacaaag gggacagagg tttacctctg gaaggatcag cgaagggtaa
     2641 agtgcaggcg gaactcagcc agcccttgac agcaggagga gcaatctcag aactgttacg
     2701 gatatctaga tttgggccca attcctgcag gcgagctctc gagccaacca ctgaggatct
     2761 gtactttcag agcgataacg atggatccga aatcggtact ggctttccat tcgaccccca
     2821 ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac
     2881 ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc
     2941 gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga
     3001 caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga
     3061 agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt
     3121 ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg
     3181 ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg
     3241 caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct
     3301 gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt
     3361 cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg
     3421 tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc
     3481 tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc
     3541 tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtctgaatct
     3601 gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtctactct
     3661 ggagatttcc ggttaataga attggcatgc aagctgatcc ggctgctaac aaagcccgaa
     3721 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcgg
     3781 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3841 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3901 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3961 ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg taaaatcgaa
     4021 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     4081 tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa ataacctctg
     4141 aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     4201 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4261 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4321 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4381 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4441 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4501 ttggaggcct aggcttttgc aaaaagcttg attcttctga cacaacagtc tcgaacttaa
     4561 ggctagagcc accatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4621 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4681 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4741 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4801 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4861 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4921 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4981 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5041 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5101 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5161 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5221 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5281 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5341 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5401 gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt cattacatct
     5461 gtgtgttggt tttttgtgtg aatcgatagc gataaggatc ctctttgcgc ttgcgttttc
     5521 ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact
     5581 ggctttctac gtaatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5641 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5701 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5761 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5821 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5881 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5941 gagcactttc aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6001 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6061 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6121 gagtgataac actgcggcca acttacttct gacaactatc ggaggaccga aggagctaac
     6181 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6241 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6301 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6361 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6421 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6481 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6541 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6601 attcgaaatg accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata
     6661 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6721 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6781 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6841 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6901 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6961 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7021 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7081 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7141 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     7201 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7261 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7321 attacgcgca gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac
     7381 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7441 ttcacctaga tccttttata gtccggaaat acaggaacgc acgctggatg gcccttcgct
     7501 gggatggtga aaccatgaaa aatggcagct tcagtggatt aagtgggggt aatgtggcct
     7561 gtaccctctg gttgcatagg tattcatacg gttaaaattt atcaggcgcg attgcggcag
     7621 tttttcgggt ggtttgttgc catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt
     7681 tttagcggtg cgtacaatta agggattatg gtaaatccac ttactgtctg ccctcgtagc
     7741 catcgagata aaccgcagta ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
