LOCUS       sequence-423370-        7183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human RBBP4. N-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7183)
  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 7183)
  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: pNLF1-N_RBBP4:1-425
FEATURES             Location/Qualifiers
     source          1..7183
                     /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             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(2951..2969)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3001..3122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3012..3031)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3066..3085
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3367..3724
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3367..3558
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3390..3410)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3575..3710
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3755..4792
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4771..4779
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4816..4864
                     /note="synthetic polyadenylation signal"
     promoter        5020..5124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5125..5193,5194..5985)
                     /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(5343..5362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6143..6731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6223..6242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6847..7130
                     /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 ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgcttc cgaattcatg gccgacaagg
     1621 aagcagcctt cgacgacgca gtggaagaac gagtgatcaa cgaggaatac aaaatatgga
     1681 aaaagaacac cccttttctt tatgatttgg tgatgaccca tgctctggag tggcccagcc
     1741 taactgccca gtggcttcca gatgtaacca gaccagaagg gaaagatttc agcattcatc
     1801 gacttgtcct ggggacacac acatcggatg aacaaaacca tcttgttata gccagtgtgc
     1861 agctccctaa tgatgatgct cagtttgatg cgtcacacta cgacagtgag aaaggagaat
     1921 ttggaggttt tggttcagtt agtggaaaaa ttgaaataga aatcaagatc aaccatgaag
     1981 gagaagtaaa cagggcccgt tatatgcccc agaacccttg tatcatcgca acaaagactc
     2041 cttccagtga tgttcttgtc tttgactata caaaacatcc ttctaaacca gatccttctg
     2101 gagagtgcaa cccagacttg cgtctccgtg gacatcagaa ggaaggctat gggctttctt
     2161 ggaacccaaa tctcagtggg cacttactta gtgcttcaga tgaccatacc atctgcctgt
     2221 gggacatcag tgccgttcca aaggagggaa aagtggtaga tgcgaagacc atctttacag
     2281 ggcatacggc agtagtagaa gatgtttcct ggcatctact ccatgagtct ctgtttgggt
     2341 cagttgctga tgatcagaaa cttatgattt gggatactcg ttcaaacaat acttccaaac
     2401 caagccactc agttgatgct cacactgctg aagtgaactg cctttctttc aatccttata
     2461 gtgagttcat tcttgccaca ggatcagctg acaagactgt tgccttgtgg gatctgagaa
     2521 atctgaaact taagttgcat tcctttgagt cacataagga tgaaatattc caggttcagt
     2581 ggtcacctca caatgagact attttagctt ccagtggtac tgatcgcaga ctgaatgtct
     2641 gggatttaag taaaattgga gaggaacaat ccccagaaga tgcagaagac gggccaccag
     2701 agttgttgtt tattcatggt ggtcatactg ccaagatatc tgatttctcc tggaatccca
     2761 atgaaccttg ggtgatttgt tctgtatcag aagacaatat catgcaagtg tggcaaatgg
     2821 cagagaacat ttataatgat gaagaccctg aaggaagcgt ggatccagaa ggacaagggt
     2881 cctgagatat ctagatttgg gcccaattcc tgcaggattt tgcggccgct tgctgagttg
     2941 gctgctgcca ccgctgagca ataactagca taaccccttg gccgcttcga gcagacatga
     3001 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3061 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3121 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     3181 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg
     3241 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3301 tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     3361 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3421 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3481 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3541 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3601 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3661 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3721 caaaaagctc gattcttctg acactagcgc caccatgaag aagcccgaac tcaccgctac
     3781 cagcgttgaa aaatttctca tcgagaagtt cgacagtgtg agcgacctga tgcagttgtc
     3841 ggagggcgaa gagagccgag ccttcagctt cgatgtcggc ggacgcggct atgtactgcg
     3901 ggtgaatagc tgcgctgatg gcttctacaa agaccgctac gtgtaccgcc acttcgccag
     3961 cgctgcacta cccatccccg aagtgttgga catcggcgag ttcagcgaga gcctgacata
     4021 ctgcatcagt agacgcgccc aaggcgttac tctccaagac ctccccgaaa cagagctgcc
     4081 tgctgtgtta cagcctgtcg ccgaagctat ggatgctatt gccgccgccg acctcagtca
     4141 aaccagcggc ttcggcccat tcgggcccca aggcatcggc cagtacacaa cctggcggga
     4201 tttcatttgc gccattgctg atccccatgt ctaccactgg cagaccgtga tggacgacac
     4261 cgtgtccgcc agcgtagctc aagccctgga cgaactgatg ctgtgggccg aagactgtcc
     4321 cgaggtgcgc cacctcgtcc atgccgactt cggcagcaac aacgtcctga ccgacaacgg
     4381 ccgcatcacc gccgtaatcg actggtccga agctatgttc ggggacagtc agtacgaggt
     4441 ggccaacatc ttcttctggc ggccctggct ggcttgcatg gagcagcaga ctcgctactt
     4501 cgagcgccgg catcccgagc tggccggcag ccctcgtctg cgagcctaca tgctgcgcat
     4561 cggcctggat cagctctacc agagcctcgt ggacggcaac ttcgacgatg ctgcctgggc
     4621 tcaaggccgc tgcgatgcca tcgtccgcag cggggccggc accgtcggtc gcacacaaat
     4681 cgctcgccgg agcgcagccg tatggaccga cggctgcgtc gaggtgctgg ccgacagcgg
     4741 caaccgccgg cccagtacac gaccgcgcgc taaggaggta ggtcgagttt agactctagc
     4801 catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     4861 tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc
     4921 cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtaatg
     4981 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5041 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5101 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5161 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5221 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5281 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac tttcaaagtt
     5341 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5401 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5461 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5521 gccaacttac ttctgacaac tatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5581 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5641 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5701 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5761 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5821 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5881 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5941 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaattcga aatgaccgac
     6001 caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6061 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6121 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6181 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6241 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6301 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6361 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6421 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6481 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6541 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     6601 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6661 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6721 aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6781 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6841 tatagtccgg aaatacagga acgcacgctg gatggccctt cgctgggatg gtgaaaccat
     6901 gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc tctggttgca
     6961 taggtattca tacggttaaa atttatcagg cgcgattgcg gcagtttttc gggtggtttg
     7021 ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca
     7081 attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga gataaaccgc
     7141 agtactccgg ccacgatgcg tccggcgtag aggatcgaga tct
//
