LOCUS       sequence-437053-        6168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DCAF8L2. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6168)
  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 6168)
  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_DCAF8L2:1-631
FEATURES             Location/Qualifiers
     source          1..6168
                     /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    3231..3352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3242..3261)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3467..3500)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3510..3557
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3589..4383
                     /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(3643..3662)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4253..4272
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4541..5129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4621..4640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5255..5303
                     /note="synthetic polyadenylation signal"
     CDS             complement(5327..5725)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5722..5731
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5756..6113)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5770..5905
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5922..6113
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6070..6090
                     /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 tgtcccacca
     1261 agaaggcagc acagacggct taccagactt agggactgaa agcctgttca gcagcccaga
     1321 ggagcagtct ggagccgtgg cggcgacaga ggcctcctca gacattgaca tagcgacctc
     1381 agagctgagt gtgacagtga ccggagatgg cagtgatagc agggatggtg gattccccaa
     1441 cgatgccagc acagaaaatc gaagctcaga ccaagaaagc gcaagtgaag acatcgaact
     1501 tgagagcttg gaggactttg agcatttcct catgagtggt gaaagtttat tccattaccc
     1561 tttagtggga gaggaggaga cagaaaggga ggaggaagac gaagagatac aagaggaggg
     1621 aggggaggag gaggaagagg aggaggagga ggaggaggag gaggaggagg aggaagaaga
     1681 acagcctcgg gcgggtccac aaggcagtgg cggcaaccat gagcagtatt cgttagagga
     1741 ggatcaggcg ctggaggagt gggtttcctc agagacatct gccctgcccc gacctcgctg
     1801 gcaggtcgtt actgctcttc accagcggca gctgggttca cgtccccgct ttgtatatga
     1861 ggcctgtggg gcaagagcct ttgtgcagcg tttccgcctg caatatcgtc ttgcagacca
     1921 tgtcggctgt gtcaatactg tacactttaa ccagcgtggc acccggctgg ccagtagcgg
     1981 tgatgaccta aaggtgatag tgtgggactg ggtgcggcag aggccagtac tgaactttga
     2041 aagtggtcac acaaataatg tcttccaggc caagttcctt cctaactgtg gtgattccac
     2101 tctggccatg tgtgcccgtg atgggcaggt acgggtagca gagttaatta atgcatcata
     2161 tttcaacaat actaagtgtg tggcccagca caggggacct gcccacaagt tggctctgga
     2221 gcctgactct ccttataagt tcctcacttc aggtgaagat gctgttgtct tcaccattga
     2281 cctcagacaa gaccggccag cttcaaaagt tgtggtaaca agagaaaatg ataagaaagt
     2341 gggactgtat acaattactg tgaatcccgc caatacctac caatttgcag tgggtggaca
     2401 agatcagttt gtaaggattt atgaccagag gaaaattgat aagaaagaaa acaatggcgt
     2461 gctcaagaaa ttcactcctc atcatctggt taattgtgat ttcccaacaa acatcacctg
     2521 cgttgtgtac agccacgatg gcacagagct gctagccagc tacaatgatg acgatattta
     2581 cctcttcaac tcctctcaca gtgatggtgc tcaatacagt aagagattta agggacacag
     2641 aaataatacc acagtcaaag gtgttaattt ctatggcccc aggagtgagt ttgtagtgag
     2701 cggtagtgat tgcgggcaca tcttcttctg ggagaaatca tcctgccaaa tcatccagtt
     2761 cctaaagggg agcagagaag gtacaataaa ctgtcttgaa ccccaccctt acctacctgt
     2821 gttggcgtgc agtggcctag atcatgatgt caagatctgg acacccacag ctaaagctgc
     2881 cactgagctt actgggttaa agaaggtgat taagaagaac aagtgggaac gagatgaaga
     2941 cagcttgcac catggcagcc tgtttgacca gtacatgctt tggttcctcc tgcgtcacgt
     3001 gacgcagaga ggtcgtcacc aggactggag aagtggtgaa gctgaatttc cagatgaaga
     3061 atcggatgag tcttccagca cttcagagac atctgaggag gaggtccaag accgagtgca
     3121 gtgcatgcca tcctaatcta gaggtcaagc ttcgggggtt taaacctgca ggcatgcaag
     3181 ctgatttggc tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat
     3241 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3301 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3361 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa
     3421 gtaaaacctc tacaaatgtg gtaaaatctt aattaagact tctaatataa cttcgtatag
     3481 catacattat acgaagttat ggttccactg ttgacaatta atcatcggca tagtatatcg
     3541 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     3601 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3661 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3721 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3781 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3841 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3901 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3961 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4021 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4081 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4141 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4201 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4261 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4321 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4381 tgattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa
     4441 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4501 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4561 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4621 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4681 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4741 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4801 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4861 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4921 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4981 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5041 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5101 agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg
     5161 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5221 tcttcaccta gatcctttta tagtccggaa tcgtcacaca aaaaaccaac acacagatgt
     5281 aatgaaaata aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca
     5341 cataaccaga gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct
     5401 tcactatggc tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg
     5461 ggctcaagat gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg
     5521 ccgcagcagc agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat
     5581 atacattgac accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt
     5641 agtcttcaga gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt
     5701 cttcttgaga caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc
     5761 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     5821 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     5881 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     5941 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     6001 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     6061 actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac
     6121 ctaaccaagt tcctctttca gaggttattt caggccatgg tgctgcgc
//
