LOCUS       sequence-425990-        7722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human BTRC. N-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7722)
  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 7722)
  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_BTRC:1-605
FEATURES             Location/Qualifiers
     source          1..7722
                     /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(3490..3508)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3540..3661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3551..3570)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3605..3624
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3906..4263
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3906..4097
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3929..3949)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4114..4249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4294..5331
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5310..5318
                     /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    5355..5403
                     /note="synthetic polyadenylation signal"
     promoter        5559..5663
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5664..5732,5733..6524)
                     /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(5882..5901)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6682..7270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6762..6781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7386..7669
                     /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 gacccggccg
     1621 aggcggtgct gcaagagaag gcactcaagt ttatgtgctc tatgcccagg tctctgtggc
     1681 tgggctgctc cagcctggcg gacagcatgc cttcgctgcg atgcctgtat aacccaggga
     1741 ctggcgcact cacagctttc cagaattcct cagagagaga agactgtaat aatggcgaac
     1801 cccctaggaa gataatacca gagaagaatt cacttagaca gacatacaac agctgtgcca
     1861 gactctgctt aaaccaagaa acagtatgtt tagcaagcac tgctatgaag actgagaatt
     1921 gtgtggccaa aacaaaactt gccaatggca cttccagtat gattgtgccc aagcaacgga
     1981 aactctcagc aagctatgaa aaggaaaagg aactgtgtgt caaatacttt gagcagtggt
     2041 cagagtcaga tcaagtggaa tttgtggaac atcttatatc ccaaatgtgt cattaccaac
     2101 atgggcacat aaactcgtat cttaaaccta tgttgcagag agatttcata actgctctgc
     2161 cagctcgggg attggatcat attgctgaga acattctgtc atacctggat gccaaatcac
     2221 tatgtgctgc tgaacttgtg tgcaaggaat ggtaccgagt gacctctgat ggcatgctgt
     2281 ggaagaagct tatcgagaga atggtcagga cagattctct gtggagaggc ctggcagaac
     2341 gaagaggatg gggacagtat ttattcaaaa acaaacctcc tgacgggaat gctcctccca
     2401 actcttttta tagagcactt tatcctaaaa ttatacaaga cattgagaca atagaatcta
     2461 attggagatg tggaagacat agtttacaga gaattcactg ccgaagtgaa acaagcaaag
     2521 gagtttactg tttacagtat gatgatcaga aaatagtaag cggccttcga gacaacacaa
     2581 tcaagatctg ggataaaaac acattggaat gcaagcgaat tctcacaggc catacaggtt
     2641 cagtcctctg tctccagtat gatgagagag tgatcataac aggatcatcg gattccacgg
     2701 tcagagtgtg ggatgtaaat acaggtgaaa tgctaaacac gttgattcac cattgtgaag
     2761 cagttctgca cttgcgtttc aataatggca tgatggtgac ctgctccaaa gatcgttcca
     2821 ttgctgtatg ggatatggcc tccccaactg acattaccct ccggagggtg ctggtcggac
     2881 accgagctgc tgtcaatgtt gtagactttg atgacaagta cattgtttct gcatctgggg
     2941 atagaactat aaaggtatgg aacacaagta cttgtgaatt tgtaaggacc ttaaatggac
     3001 acaaacgagg cattgcctgt ttgcagtaca gggacaggct ggtagtgagt ggctcatctg
     3061 acaacactat cagattatgg gacatagaat gtggtgcatg tttacgagtg ttagaaggcc
     3121 atgaggaatt ggtgcgttgt attcgatttg ataacaagag gatagtcagt ggggcctatg
     3181 atggaaaaat taaagtgtgg gatcttgtgg ctgctttgga cccccgtgct cctgcaggga
     3241 cactctgtct acggaccctt gtggagcatt ccggaagagt ttttcgacta cagtttgatg
     3301 aattccagat tgtcagtagt tcacatgatg acacaatcct catctgggac ttcctaaatg
     3361 atccagctgc ccaagctgaa cccccccgtt ccccttctcg aacatacacc tacatctcca
     3421 gataataatc tagatttggg cccaattcct gcaggatttt gcggccgctt gctgagttgg
     3481 ctgctgccac cgctgagcaa taactagcat aaccccttgg ccgcttcgag cagacatgat
     3541 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3601 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3661 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     3721 tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc
     3781 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3841 acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
     3901 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3961 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4081 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4141 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4201 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4261 aaaaagctcg attcttctga cactagcgcc accatgaaga agcccgaact caccgctacc
     4321 agcgttgaaa aatttctcat cgagaagttc gacagtgtga gcgacctgat gcagttgtcg
     4381 gagggcgaag agagccgagc cttcagcttc gatgtcggcg gacgcggcta tgtactgcgg
     4441 gtgaatagct gcgctgatgg cttctacaaa gaccgctacg tgtaccgcca cttcgccagc
     4501 gctgcactac ccatccccga agtgttggac atcggcgagt tcagcgagag cctgacatac
     4561 tgcatcagta gacgcgccca aggcgttact ctccaagacc tccccgaaac agagctgcct
     4621 gctgtgttac agcctgtcgc cgaagctatg gatgctattg ccgccgccga cctcagtcaa
     4681 accagcggct tcggcccatt cgggccccaa ggcatcggcc agtacacaac ctggcgggat
     4741 ttcatttgcg ccattgctga tccccatgtc taccactggc agaccgtgat ggacgacacc
     4801 gtgtccgcca gcgtagctca agccctggac gaactgatgc tgtgggccga agactgtccc
     4861 gaggtgcgcc acctcgtcca tgccgacttc ggcagcaaca acgtcctgac cgacaacggc
     4921 cgcatcaccg ccgtaatcga ctggtccgaa gctatgttcg gggacagtca gtacgaggtg
     4981 gccaacatct tcttctggcg gccctggctg gcttgcatgg agcagcagac tcgctacttc
     5041 gagcgccggc atcccgagct ggccggcagc cctcgtctgc gagcctacat gctgcgcatc
     5101 ggcctggatc agctctacca gagcctcgtg gacggcaact tcgacgatgc tgcctgggct
     5161 caaggccgct gcgatgccat cgtccgcagc ggggccggca ccgtcggtcg cacacaaatc
     5221 gctcgccgga gcgcagccgt atggaccgac ggctgcgtcg aggtgctggc cgacagcggc
     5281 aaccgccggc ccagtacacg accgcgcgct aaggaggtag gtcgagttta gactctagcc
     5341 atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     5401 gtgaatcgat agcgataagg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc
     5461 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtaatgg
     5521 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5581 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5641 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5701 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5761 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5821 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact ttcaaagttc
     5881 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5941 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6001 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6061 ccaacttact tctgacaact atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6121 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6181 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6241 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6301 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6361 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6421 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6481 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaattcgaa atgaccgacc
     6541 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6601 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6661 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6721 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6781 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6841 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6901 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6961 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7021 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7081 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7141 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7201 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7261 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7321 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7381 atagtccgga aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg
     7441 aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat
     7501 aggtattcat acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt
     7561 tgccattttt acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa
     7621 ttaagggatt atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca
     7681 gtactccggc cacgatgcgt ccggcgtaga ggatcgagat ct
//
