LOCUS       sequence-426004-        8097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DTL. N-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8097)
  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 8097)
  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_DTL:1-730
FEATURES             Location/Qualifiers
     source          1..8097
                     /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(3865..3883)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3915..4036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3926..3945)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3980..3999
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4281..4638
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4281..4472
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4304..4324)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4489..4624
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4669..5706
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5685..5693
                     /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    5730..5778
                     /note="synthetic polyadenylation signal"
     promoter        5934..6038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6039..6107,6108..6899)
                     /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(6257..6276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7057..7645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7137..7156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7761..8044
                     /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 ctcttcaatt
     1621 cggtgctccg ccagccccag cttggcgtcc tgagaaatgg atggtcttca caataccctc
     1681 ttcaatccct tctgactggt tatcagtgca gtggtaatga tgaacacact tcttatggag
     1741 aaacaggagt cccagttcct ccttttggat gtaccttctc ttctgctccc aatatggaac
     1801 atgtactagc agttgccaat gaagaaggct ttgttcgatt gtataacaca gaatcacaaa
     1861 gtttcagaaa gaagtgcttc aaagaatgga tggctcactg gaatgccgtc tttgacctgg
     1921 cctgggttcc tggtgaactt aaacttgtta cagcagcagg tgatcaaaca gccaaatttt
     1981 gggacgtaaa agctggtgag ctgattggaa catgcaaagg tcatcaatgc agcctcaagt
     2041 cagttgcctt ttctaagttt gagaaagctg tattctgtac gggtggaaga gatggcaaca
     2101 ttatggtctg ggataccagg tgcaacaaaa aagatgggtt ttataggcaa gtgaatcaaa
     2161 tcagtggagc tcacaatacc tcagacaagc aaaccccttc aaaacccaag aagaaacaga
     2221 attcaaaagg acttgctcct tctgtggatt tccagcaaag tgttactgtg gtcctctttc
     2281 aagacgagaa taccttagtc tcagcaggag ctgtggatgg gataatcaaa gtatgggatt
     2341 tacgtaagaa ttatactgct tatcgacaag aacccatagc atccaagtct ttcctgtacc
     2401 caggtagcag cactcgaaaa cttggatatt caagtctgat tttggattcc actggctcta
     2461 ctttatttgc taattgcaca gacgataaca tctacatgtt taatatgact gggttgaaga
     2521 cttctccagt ggctattttc aatggacacc agaactctac cttttatgta aaatccagcc
     2581 ttagtccaga tgaccagttt ttagtcagtg gctcaagtga tgaagctgcc tacatatgga
     2641 aggtctccac accctggcaa cctcctactg tgctcctggg tcattctcaa gaggtcacgt
     2701 ctgtgtgctg gtgtccatct gacttcacaa agattgctac ctgttctgat gacaatacac
     2761 taaaaatctg gcgcttgaat agaggcttag aggagaaacc aggaggtgat aaactttcca
     2821 cggtgggttg ggcctctcag aagaaaaaag agtcaagacc tggcctagta acagtaacga
     2881 gtagccagag tactcctgcc aaagccccca gggcaaagtg caatccatcc aattcttccc
     2941 cgtcatccgc agcttgtgcc ccaagctgtg ctggagacct ccctcttcct tcaaatactc
     3001 ctacgttctc tattaaaacc tctcctgcca aggcccggtc tcccatcaac agaagaggct
     3061 ctgtctcctc cgtctctccc aagccacctt catctttcaa gatgtcgatt agaaactggg
     3121 tgacccgaac accttcctca tcaccaccca tcactccacc tgcttcggag accaagatca
     3181 tgtctccgag aaaagccctt attcctgtga gccagaagtc atcccaagca gaggcttgct
     3241 ctgagtctag aaatagagta aagaggaggc tagactcaag ctgtctggag agtgtgaaac
     3301 aaaagtgtgt gaagagttgt aactgtgtga ctgagcttga tggccaagtt gaaaatcttc
     3361 atttggatct gtgctgcctt gctggtaacc aggaagacct tagtaaggac tctctaggtc
     3421 ctaccaaatc aagcaaaatt gaaggagctg gtaccagtat ctcagagcct ccgtctccta
     3481 tcagtccgta tgcttcagaa agctgtggaa cgctacctct tcctttgaga ccttgtggag
     3541 aagggtctga aatggtaggc aaagagaata gttccccaga gaataaaaac tggttgttgg
     3601 ccatggcagc caaacggaag gctgagaatc catctccacg aagtccgtca tcccagacac
     3661 ccaattccag gagacagagc ggaaagaaat tgccaagccc ggtcaccatc acgcccagct
     3721 ccatgaggaa aatctgcaca tacttccata gaaagtccca ggaggacttc tgtggtcctg
     3781 aacactcaac agagttataa taatctagat ttgggcccaa ttcctgcagg attttgcggc
     3841 cgcttgctga gttggctgct gccaccgctg agcaataact agcataaccc cttggccgct
     3901 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3961 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     4021 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     4081 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta
     4141 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     4201 tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
     4261 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4321 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4381 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4441 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4501 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4561 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4621 ggcctaggct tttgcaaaaa gctcgattct tctgacacta gcgccaccat gaagaagccc
     4681 gaactcaccg ctaccagcgt tgaaaaattt ctcatcgaga agttcgacag tgtgagcgac
     4741 ctgatgcagt tgtcggaggg cgaagagagc cgagccttca gcttcgatgt cggcggacgc
     4801 ggctatgtac tgcgggtgaa tagctgcgct gatggcttct acaaagaccg ctacgtgtac
     4861 cgccacttcg ccagcgctgc actacccatc cccgaagtgt tggacatcgg cgagttcagc
     4921 gagagcctga catactgcat cagtagacgc gcccaaggcg ttactctcca agacctcccc
     4981 gaaacagagc tgcctgctgt gttacagcct gtcgccgaag ctatggatgc tattgccgcc
     5041 gccgacctca gtcaaaccag cggcttcggc ccattcgggc cccaaggcat cggccagtac
     5101 acaacctggc gggatttcat ttgcgccatt gctgatcccc atgtctacca ctggcagacc
     5161 gtgatggacg acaccgtgtc cgccagcgta gctcaagccc tggacgaact gatgctgtgg
     5221 gccgaagact gtcccgaggt gcgccacctc gtccatgccg acttcggcag caacaacgtc
     5281 ctgaccgaca acggccgcat caccgccgta atcgactggt ccgaagctat gttcggggac
     5341 agtcagtacg aggtggccaa catcttcttc tggcggccct ggctggcttg catggagcag
     5401 cagactcgct acttcgagcg ccggcatccc gagctggccg gcagccctcg tctgcgagcc
     5461 tacatgctgc gcatcggcct ggatcagctc taccagagcc tcgtggacgg caacttcgac
     5521 gatgctgcct gggctcaagg ccgctgcgat gccatcgtcc gcagcggggc cggcaccgtc
     5581 ggtcgcacac aaatcgctcg ccggagcgca gccgtatgga ccgacggctg cgtcgaggtg
     5641 ctggccgaca gcggcaaccg ccggcccagt acacgaccgc gcgctaagga ggtaggtcga
     5701 gtttagactc tagccatcac gatggccgca ataaaatatc tttattttca ttacatctgt
     5761 gtgttggttt tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt gcgttttccc
     5821 ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg
     5881 ctttctacgt aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5941 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     6001 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6061 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6121 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6181 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6241 gcactttcaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6301 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6361 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6421 gtgataacac tgcggccaac ttacttctga caactatcgg aggaccgaag gagctaaccg
     6481 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6541 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6601 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6661 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6721 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6781 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6841 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaat
     6901 tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg
     6961 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7021 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7081 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7141 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7201 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7261 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7321 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7381 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7441 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7501 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7561 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7621 tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7681 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7741 cacctagatc cttttatagt ccggaaatac aggaacgcac gctggatggc ccttcgctgg
     7801 gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa tgtggcctgt
     7861 accctctggt tgcataggta ttcatacggt taaaatttat caggcgcgat tgcggcagtt
     7921 tttcgggtgg tttgttgcca tttttacctg tctgctgccg tgatcgcgct gaacgcgttt
     7981 tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca
     8041 tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatct
//
