LOCUS       sequence-425943-        8148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DTL. C-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8148)
  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 8148)
  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-C_DTL:1-730
FEATURES             Location/Qualifiers
     source          1..8148
                     /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             3323..3835
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3911..3929)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3966..4087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3977..3996)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4031..4050
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4332..4689
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4332..4523
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4355..4375)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4540..4675
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4720..5757
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5736..5744
                     /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    5781..5829
                     /note="synthetic polyadenylation signal"
     promoter        5985..6089
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6090..6158,6159..6950)
                     /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(6308..6327)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7108..7696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7188..7207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7812..8095
                     /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 caccatgctc ttcaattcgg tgctccgcca gccccagctt ggcgtcctga gaaatggatg
     1141 gtcttcacaa taccctcttc aatcccttct gactggttat cagtgcagtg gtaatgatga
     1201 acacacttct tatggagaaa caggagtccc agttcctcct tttggatgta ccttctcttc
     1261 tgctcccaat atggaacatg tactagcagt tgccaatgaa gaaggctttg ttcgattgta
     1321 taacacagaa tcacaaagtt tcagaaagaa gtgcttcaaa gaatggatgg ctcactggaa
     1381 tgccgtcttt gacctggcct gggttcctgg tgaacttaaa cttgttacag cagcaggtga
     1441 tcaaacagcc aaattttggg acgtaaaagc tggtgagctg attggaacat gcaaaggtca
     1501 tcaatgcagc ctcaagtcag ttgccttttc taagtttgag aaagctgtat tctgtacggg
     1561 tggaagagat ggcaacatta tggtctggga taccaggtgc aacaaaaaag atgggtttta
     1621 taggcaagtg aatcaaatca gtggagctca caatacctca gacaagcaaa ccccttcaaa
     1681 acccaagaag aaacagaatt caaaaggact tgctccttct gtggatttcc agcaaagtgt
     1741 tactgtggtc ctctttcaag acgagaatac cttagtctca gcaggagctg tggatgggat
     1801 aatcaaagta tgggatttac gtaagaatta tactgcttat cgacaagaac ccatagcatc
     1861 caagtctttc ctgtacccag gtagcagcac tcgaaaactt ggatattcaa gtctgatttt
     1921 ggattccact ggctctactt tatttgctaa ttgcacagac gataacatct acatgtttaa
     1981 tatgactggg ttgaagactt ctccagtggc tattttcaat ggacaccaga actctacctt
     2041 ttatgtaaaa tccagcctta gtccagatga ccagttttta gtcagtggct caagtgatga
     2101 agctgcctac atatggaagg tctccacacc ctggcaacct cctactgtgc tcctgggtca
     2161 ttctcaagag gtcacgtctg tgtgctggtg tccatctgac ttcacaaaga ttgctacctg
     2221 ttctgatgac aatacactaa aaatctggcg cttgaataga ggcttagagg agaaaccagg
     2281 aggtgataaa ctttccacgg tgggttgggc ctctcagaag aaaaaagagt caagacctgg
     2341 cctagtaaca gtaacgagta gccagagtac tcctgccaaa gcccccaggg caaagtgcaa
     2401 tccatccaat tcttccccgt catccgcagc ttgtgcccca agctgtgctg gagacctccc
     2461 tcttccttca aatactccta cgttctctat taaaacctct cctgccaagg cccggtctcc
     2521 catcaacaga agaggctctg tctcctccgt ctctcccaag ccaccttcat ctttcaagat
     2581 gtcgattaga aactgggtga cccgaacacc ttcctcatca ccacccatca ctccacctgc
     2641 ttcggagacc aagatcatgt ctccgagaaa agcccttatt cctgtgagcc agaagtcatc
     2701 ccaagcagag gcttgctctg agtctagaaa tagagtaaag aggaggctag actcaagctg
     2761 tctggagagt gtgaaacaaa agtgtgtgaa gagttgtaac tgtgtgactg agcttgatgg
     2821 ccaagttgaa aatcttcatt tggatctgtg ctgccttgct ggtaaccagg aagaccttag
     2881 taaggactct ctaggtccta ccaaatcaag caaaattgaa ggagctggta ccagtatctc
     2941 agagcctccg tctcctatca gtccgtatgc ttcagaaagc tgtggaacgc tacctcttcc
     3001 tttgagacct tgtggagaag ggtctgaaat ggtaggcaaa gagaatagtt ccccagagaa
     3061 taaaaactgg ttgttggcca tggcagccaa acggaaggct gagaatccat ctccacgaag
     3121 tccgtcatcc cagacaccca attccaggag acagagcgga aagaaattgc caagcccggt
     3181 caccatcacg cccagctcca tgaggaaaat ctgcacatac ttccatagaa agtcccagga
     3241 ggacttctgt ggtcctgaac actcaacaga gttatctaga tttgggccca attcctgcag
     3301 gcgagctctc ggctcgagcg gcgtcttcac actcgaagat ttcgttgggg actggcgaca
     3361 gacagccggc tacaacctgg accaagtcct tgaacaggga ggtgtgtcca gtttgtttca
     3421 gaatctcggg gtgtccgtaa ctccgatcca aaggattgtc ctgagcggtg aaaatgggct
     3481 gaagatcgac atccatgtca tcatcccgta tgaaggtctg agcggcgacc aaatgggcca
     3541 gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat catcacttta aggtgatcct
     3601 gcactatggc acactggtaa tcgacggggt tacgccgaac atgatcgact atttcggacg
     3661 gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc actgtaacag ggaccctgtg
     3721 gaacggcaac aaaattatcg acgagcgcct gatcaacccc gacggctccc tgctgttccg
     3781 agtaaccatc aacggagtga ccggctggcg gctgtgcgaa cgcattctgg cgtaaggccg
     3841 cgactctagg gcatgcaagc tgatccggct gctaacaaag cccgaaagga agctgagttg
     3901 gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc ttcgagcaga
     3961 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4021 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4081 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     4141 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat
     4201 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4261 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     4321 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     4381 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4441 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4501 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4561 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4621 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4681 ttttgcaaaa agctcgattc ttctgacact agcgccacca tgaagaagcc cgaactcacc
     4741 gctaccagcg ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga cctgatgcag
     4801 ttgtcggagg gcgaagagag ccgagccttc agcttcgatg tcggcggacg cggctatgta
     4861 ctgcgggtga atagctgcgc tgatggcttc tacaaagacc gctacgtgta ccgccacttc
     4921 gccagcgctg cactacccat ccccgaagtg ttggacatcg gcgagttcag cgagagcctg
     4981 acatactgca tcagtagacg cgcccaaggc gttactctcc aagacctccc cgaaacagag
     5041 ctgcctgctg tgttacagcc tgtcgccgaa gctatggatg ctattgccgc cgccgacctc
     5101 agtcaaacca gcggcttcgg cccattcggg ccccaaggca tcggccagta cacaacctgg
     5161 cgggatttca tttgcgccat tgctgatccc catgtctacc actggcagac cgtgatggac
     5221 gacaccgtgt ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg ggccgaagac
     5281 tgtcccgagg tgcgccacct cgtccatgcc gacttcggca gcaacaacgt cctgaccgac
     5341 aacggccgca tcaccgccgt aatcgactgg tccgaagcta tgttcgggga cagtcagtac
     5401 gaggtggcca acatcttctt ctggcggccc tggctggctt gcatggagca gcagactcgc
     5461 tacttcgagc gccggcatcc cgagctggcc ggcagccctc gtctgcgagc ctacatgctg
     5521 cgcatcggcc tggatcagct ctaccagagc ctcgtggacg gcaacttcga cgatgctgcc
     5581 tgggctcaag gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt cggtcgcaca
     5641 caaatcgctc gccggagcgc agccgtatgg accgacggct gcgtcgaggt gctggccgac
     5701 agcggcaacc gccggcccag tacacgaccg cgcgctaagg aggtaggtcg agtttagact
     5761 ctagccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     5821 ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga
     5881 tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg
     5941 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6001 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6061 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6121 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6181 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6241 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttca
     6301 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6361 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6421 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6481 ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc gcttttttgc
     6541 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6601 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6661 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6721 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6781 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6841 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6901 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ttcgaaatga
     6961 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7021 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7081 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7141 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7201 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7261 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7321 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7381 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7441 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7501 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7561 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7621 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7681 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7741 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7801 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     7861 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     7921 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     7981 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     8041 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     8101 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
