LOCUS       sequence-425986-        6474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DTL. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6474)
  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 6474)
  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-N_DTL:1-730
FEATURES             Location/Qualifiers
     source          1..6474
                     /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    3537..3658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3548..3567)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3773..3806)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3816..3863
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3895..4689
                     /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(3949..3968)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4559..4578
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4847..5435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4927..4946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5561..5609
                     /note="synthetic polyadenylation signal"
     CDS             complement(5633..6031)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6028..6037
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6062..6419)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6076..6211
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6228..6419
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6376..6396
                     /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 ataggggtta gcgatcgcca tggtgagcgg
     1201 ctggcggctg ttcaagaaga ttagcgggag ctccggtggc tcgagcggta tgctcttcaa
     1261 ttcggtgctc cgccagcccc agcttggcgt cctgagaaat ggatggtctt cacaataccc
     1321 tcttcaatcc cttctgactg gttatcagtg cagtggtaat gatgaacaca cttcttatgg
     1381 agaaacagga gtcccagttc ctccttttgg atgtaccttc tcttctgctc ccaatatgga
     1441 acatgtacta gcagttgcca atgaagaagg ctttgttcga ttgtataaca cagaatcaca
     1501 aagtttcaga aagaagtgct tcaaagaatg gatggctcac tggaatgccg tctttgacct
     1561 ggcctgggtt cctggtgaac ttaaacttgt tacagcagca ggtgatcaaa cagccaaatt
     1621 ttgggacgta aaagctggtg agctgattgg aacatgcaaa ggtcatcaat gcagcctcaa
     1681 gtcagttgcc ttttctaagt ttgagaaagc tgtattctgt acgggtggaa gagatggcaa
     1741 cattatggtc tgggatacca ggtgcaacaa aaaagatggg ttttataggc aagtgaatca
     1801 aatcagtgga gctcacaata cctcagacaa gcaaacccct tcaaaaccca agaagaaaca
     1861 gaattcaaaa ggacttgctc cttctgtgga tttccagcaa agtgttactg tggtcctctt
     1921 tcaagacgag aataccttag tctcagcagg agctgtggat gggataatca aagtatggga
     1981 tttacgtaag aattatactg cttatcgaca agaacccata gcatccaagt ctttcctgta
     2041 cccaggtagc agcactcgaa aacttggata ttcaagtctg attttggatt ccactggctc
     2101 tactttattt gctaattgca cagacgataa catctacatg tttaatatga ctgggttgaa
     2161 gacttctcca gtggctattt tcaatggaca ccagaactct accttttatg taaaatccag
     2221 ccttagtcca gatgaccagt ttttagtcag tggctcaagt gatgaagctg cctacatatg
     2281 gaaggtctcc acaccctggc aacctcctac tgtgctcctg ggtcattctc aagaggtcac
     2341 gtctgtgtgc tggtgtccat ctgacttcac aaagattgct acctgttctg atgacaatac
     2401 actaaaaatc tggcgcttga atagaggctt agaggagaaa ccaggaggtg ataaactttc
     2461 cacggtgggt tgggcctctc agaagaaaaa agagtcaaga cctggcctag taacagtaac
     2521 gagtagccag agtactcctg ccaaagcccc cagggcaaag tgcaatccat ccaattcttc
     2581 cccgtcatcc gcagcttgtg ccccaagctg tgctggagac ctccctcttc cttcaaatac
     2641 tcctacgttc tctattaaaa cctctcctgc caaggcccgg tctcccatca acagaagagg
     2701 ctctgtctcc tccgtctctc ccaagccacc ttcatctttc aagatgtcga ttagaaactg
     2761 ggtgacccga acaccttcct catcaccacc catcactcca cctgcttcgg agaccaagat
     2821 catgtctccg agaaaagccc ttattcctgt gagccagaag tcatcccaag cagaggcttg
     2881 ctctgagtct agaaatagag taaagaggag gctagactca agctgtctgg agagtgtgaa
     2941 acaaaagtgt gtgaagagtt gtaactgtgt gactgagctt gatggccaag ttgaaaatct
     3001 tcatttggat ctgtgctgcc ttgctggtaa ccaggaagac cttagtaagg actctctagg
     3061 tcctaccaaa tcaagcaaaa ttgaaggagc tggtaccagt atctcagagc ctccgtctcc
     3121 tatcagtccg tatgcttcag aaagctgtgg aacgctacct cttcctttga gaccttgtgg
     3181 agaagggtct gaaatggtag gcaaagagaa tagttcccca gagaataaaa actggttgtt
     3241 ggccatggca gccaaacgga aggctgagaa tccatctcca cgaagtccgt catcccagac
     3301 acccaattcc aggagacaga gcggaaagaa attgccaagc ccggtcacca tcacgcccag
     3361 ctccatgagg aaaatctgca catacttcca tagaaagtcc caggaggact tctgtggtcc
     3421 tgaacactca acagagttat aatctagagg tcaagcttcg ggggtttaaa cctgcaggca
     3481 tgcaagctga tttggctgct gccaccgctg agcaaggccg cttcgagcag acatgataag
     3541 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3601 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3661 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt
     3721 aagcaagtaa aacctctaca aatgtggtaa aatcttaatt aagacttcta atataacttc
     3781 gtatagcata cattatacga agttatggtt ccactgttga caattaatca tcggcatagt
     3841 atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt
     3901 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3961 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4021 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4081 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4141 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4201 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4261 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4321 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4381 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4441 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4501 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4561 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4621 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4681 ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
     4741 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4801 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4861 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4921 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4981 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5041 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5101 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5161 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5221 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5281 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5341 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5401 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     5461 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5521 aaaggatctt cacctagatc cttttatagt ccggaatcgt cacacaaaaa accaacacac
     5581 agatgtaatg aaaataaaga tattttattg cggccatcgt gatggctaga gtttagccct
     5641 cccacacata accagagggc agcaattcac gaatcccaac tgccgtcggc tgtccatcac
     5701 tgtccttcac tatggctttg atcccaggat gcagatcgag aagcacctgt cggcaccgtc
     5761 cgcaggggct caagatgccc ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc
     5821 cagctgccgc agcagcagca gtgcccagca ccacgagttc tgcacaaggt cccccagtaa
     5881 aatgatatac attgacacca gtgaagatgc ggccgtcgct agagagagct gcgctggcga
     5941 cgctgtagtc ttcagagatg gggatgctgt tgattgtagc cgttgctctt tcaatgaggg
     6001 tggattcttc ttgagacaaa ggcttggcca tggtggcgct agtgtcagaa gaatcgagct
     6061 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     6121 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     6181 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
     6241 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     6301 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     6361 ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga
     6421 aggtacctaa ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgc
//
