LOCUS       sequence-425976-        6518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DTL. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6518)
  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 6518)
  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: pBiT3.1-C_DTL:1-730
FEATURES             Location/Qualifiers
     source          1..6518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3581..3702
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3592..3611)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3817..3850)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3860..3907
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3939..4733
                     /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(3993..4012)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4603..4622
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4891..5479)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4971..4990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5605..5653
                     /note="synthetic polyadenylation signal"
     CDS             complement(5677..6075)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6072..6081
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6106..6463)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6120..6255
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6272..6463
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6420..6440
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggcga tcgcaacaag cttcgggaag
     1201 atctggagct agcggcgggg gatccggtgg gaattcgaca ccatgctctt caattcggtg
     1261 ctccgccagc cccagcttgg cgtcctgaga aatggatggt cttcacaata ccctcttcaa
     1321 tcccttctga ctggttatca gtgcagtggt aatgatgaac acacttctta tggagaaaca
     1381 ggagtcccag ttcctccttt tggatgtacc ttctcttctg ctcccaatat ggaacatgta
     1441 ctagcagttg ccaatgaaga aggctttgtt cgattgtata acacagaatc acaaagtttc
     1501 agaaagaagt gcttcaaaga atggatggct cactggaatg ccgtctttga cctggcctgg
     1561 gttcctggtg aacttaaact tgttacagca gcaggtgatc aaacagccaa attttgggac
     1621 gtaaaagctg gtgagctgat tggaacatgc aaaggtcatc aatgcagcct caagtcagtt
     1681 gccttttcta agtttgagaa agctgtattc tgtacgggtg gaagagatgg caacattatg
     1741 gtctgggata ccaggtgcaa caaaaaagat gggttttata ggcaagtgaa tcaaatcagt
     1801 ggagctcaca atacctcaga caagcaaacc ccttcaaaac ccaagaagaa acagaattca
     1861 aaaggacttg ctccttctgt ggatttccag caaagtgtta ctgtggtcct ctttcaagac
     1921 gagaatacct tagtctcagc aggagctgtg gatgggataa tcaaagtatg ggatttacgt
     1981 aagaattata ctgcttatcg acaagaaccc atagcatcca agtctttcct gtacccaggt
     2041 agcagcactc gaaaacttgg atattcaagt ctgattttgg attccactgg ctctacttta
     2101 tttgctaatt gcacagacga taacatctac atgtttaata tgactgggtt gaagacttct
     2161 ccagtggcta ttttcaatgg acaccagaac tctacctttt atgtaaaatc cagccttagt
     2221 ccagatgacc agtttttagt cagtggctca agtgatgaag ctgcctacat atggaaggtc
     2281 tccacaccct ggcaacctcc tactgtgctc ctgggtcatt ctcaagaggt cacgtctgtg
     2341 tgctggtgtc catctgactt cacaaagatt gctacctgtt ctgatgacaa tacactaaaa
     2401 atctggcgct tgaatagagg cttagaggag aaaccaggag gtgataaact ttccacggtg
     2461 ggttgggcct ctcagaagaa aaaagagtca agacctggcc tagtaacagt aacgagtagc
     2521 cagagtactc ctgccaaagc ccccagggca aagtgcaatc catccaattc ttccccgtca
     2581 tccgcagctt gtgccccaag ctgtgctgga gacctccctc ttccttcaaa tactcctacg
     2641 ttctctatta aaacctctcc tgccaaggcc cggtctccca tcaacagaag aggctctgtc
     2701 tcctccgtct ctcccaagcc accttcatct ttcaagatgt cgattagaaa ctgggtgacc
     2761 cgaacacctt cctcatcacc acccatcact ccacctgctt cggagaccaa gatcatgtct
     2821 ccgagaaaag cccttattcc tgtgagccag aagtcatccc aagcagaggc ttgctctgag
     2881 tctagaaata gagtaaagag gaggctagac tcaagctgtc tggagagtgt gaaacaaaag
     2941 tgtgtgaaga gttgtaactg tgtgactgag cttgatggcc aagttgaaaa tcttcatttg
     3001 gatctgtgct gccttgctgg taaccaggaa gaccttagta aggactctct aggtcctacc
     3061 aaatcaagca aaattgaagg agctggtacc agtatctcag agcctccgtc tcctatcagt
     3121 ccgtatgctt cagaaagctg tggaacgcta cctcttcctt tgagaccttg tggagaaggg
     3181 tctgaaatgg taggcaaaga gaatagttcc ccagagaata aaaactggtt gttggccatg
     3241 gcagccaaac ggaaggctga gaatccatct ccacgaagtc cgtcatccca gacacccaat
     3301 tccaggagac agagcggaaa gaaattgcca agcccggtca ccatcacgcc cagctccatg
     3361 aggaaaatct gcacatactt ccatagaaag tcccaggagg acttctgtgg tcctgaacac
     3421 tcaacagagt taggctcgag cggtgggagc tccggtgtga gcggctggcg gctgttcaag
     3481 aagattagct aagtttaaac ggccgcgact ctagactgca ggcatgcaag ctgatttggc
     3541 tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat tgatgagttt
     3601 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3661 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3721 cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc
     3781 tacaaatgtg gtaaaatctt aattaagact tctaatataa cttcgtatag catacattat
     3841 acgaagttat ggttccactg ttgacaatta atcatcggca tagtatatcg gcatagtata
     3901 atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc
     3961 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4021 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4081 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4141 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4201 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4261 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4321 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4381 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4441 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4501 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4561 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4621 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4681 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgattcgaaa
     4741 tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc
     4801 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4861 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4921 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4981 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5041 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5101 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5161 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5221 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5281 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5341 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5401 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5461 cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5521 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5581 gatcctttta tagtccggaa tcgtcacaca aaaaaccaac acacagatgt aatgaaaata
     5641 aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca cataaccaga
     5701 gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct tcactatggc
     5761 tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat
     5821 gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc
     5881 agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat atacattgac
     5941 accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt agtcttcaga
     6001 gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt cttcttgaga
     6061 caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc aaaagcctag
     6121 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     6181 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     6241 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     6301 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     6361 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     6421 ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac ctaaccaagt
     6481 tcctctttca gaggttattt caggccatgg tgctgcgc
//
