LOCUS       sequence-423460-        6212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DCAF8L2. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6212)
  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 6212)
  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_DCAF8L2:1-631
FEATURES             Location/Qualifiers
     source          1..6212
                     /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    3275..3396
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3286..3305)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3511..3544)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3554..3601
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3633..4427
                     /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(3687..3706)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4297..4316
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4585..5173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4665..4684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5299..5347
                     /note="synthetic polyadenylation signal"
     CDS             complement(5371..5769)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5766..5775
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5800..6157)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5814..5949
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5966..6157
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6114..6134
                     /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 ccatgtccca ccaagaaggc
     1261 agcacagacg gcttaccaga cttagggact gaaagcctgt tcagcagccc agaggagcag
     1321 tctggagccg tggcggcgac agaggcctcc tcagacattg acatagcgac ctcagagctg
     1381 agtgtgacag tgaccggaga tggcagtgat agcagggatg gtggattccc caacgatgcc
     1441 agcacagaaa atcgaagctc agaccaagaa agcgcaagtg aagacatcga acttgagagc
     1501 ttggaggact ttgagcattt cctcatgagt ggtgaaagtt tattccatta ccctttagtg
     1561 ggagaggagg agacagaaag ggaggaggaa gacgaagaga tacaagagga gggaggggag
     1621 gaggaggaag aggaggagga ggaggaggag gaggaggagg aggaggaaga agaacagcct
     1681 cgggcgggtc cacaaggcag tggcggcaac catgagcagt attcgttaga ggaggatcag
     1741 gcgctggagg agtgggtttc ctcagagaca tctgccctgc cccgacctcg ctggcaggtc
     1801 gttactgctc ttcaccagcg gcagctgggt tcacgtcccc gctttgtata tgaggcctgt
     1861 ggggcaagag cctttgtgca gcgtttccgc ctgcaatatc gtcttgcaga ccatgtcggc
     1921 tgtgtcaata ctgtacactt taaccagcgt ggcacccggc tggccagtag cggtgatgac
     1981 ctaaaggtga tagtgtggga ctgggtgcgg cagaggccag tactgaactt tgaaagtggt
     2041 cacacaaata atgtcttcca ggccaagttc cttcctaact gtggtgattc cactctggcc
     2101 atgtgtgccc gtgatgggca ggtacgggta gcagagttaa ttaatgcatc atatttcaac
     2161 aatactaagt gtgtggccca gcacagggga cctgcccaca agttggctct ggagcctgac
     2221 tctccttata agttcctcac ttcaggtgaa gatgctgttg tcttcaccat tgacctcaga
     2281 caagaccggc cagcttcaaa agttgtggta acaagagaaa atgataagaa agtgggactg
     2341 tatacaatta ctgtgaatcc cgccaatacc taccaatttg cagtgggtgg acaagatcag
     2401 tttgtaagga tttatgacca gaggaaaatt gataagaaag aaaacaatgg cgtgctcaag
     2461 aaattcactc ctcatcatct ggttaattgt gatttcccaa caaacatcac ctgcgttgtg
     2521 tacagccacg atggcacaga gctgctagcc agctacaatg atgacgatat ttacctcttc
     2581 aactcctctc acagtgatgg tgctcaatac agtaagagat ttaagggaca cagaaataat
     2641 accacagtca aaggtgttaa tttctatggc cccaggagtg agtttgtagt gagcggtagt
     2701 gattgcgggc acatcttctt ctgggagaaa tcatcctgcc aaatcatcca gttcctaaag
     2761 gggagcagag aaggtacaat aaactgtctt gaaccccacc cttacctacc tgtgttggcg
     2821 tgcagtggcc tagatcatga tgtcaagatc tggacaccca cagctaaagc tgccactgag
     2881 cttactgggt taaagaaggt gattaagaag aacaagtggg aacgagatga agacagcttg
     2941 caccatggca gcctgtttga ccagtacatg ctttggttcc tcctgcgtca cgtgacgcag
     3001 agaggtcgtc accaggactg gagaagtggt gaagctgaat ttccagatga agaatcggat
     3061 gagtcttcca gcacttcaga gacatctgag gaggaggtcc aagaccgagt gcagtgcatg
     3121 ccatccggct cgagcggtgg gagctccggt gtgagcggct ggcggctgtt caagaagatt
     3181 agctaagttt aaacggccgc gactctagac tgcaggcatg caagctgatt tggctgctgc
     3241 caccgctgag caaggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     3301 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     3361 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     3421 atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa
     3481 tgtggtaaaa tcttaattaa gacttctaat ataacttcgt atagcataca ttatacgaag
     3541 ttatggttcc actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     3601 caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag
     3661 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3721 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3781 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3841 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3901 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3961 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4021 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4081 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4141 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4201 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4261 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4321 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4381 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc gaaatgaccg
     4441 accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4501 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4561 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4621 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4681 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4741 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4801 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4861 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4921 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4981 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     5041 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5101 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5161 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5221 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5281 tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa aataaagata
     5341 ttttattgcg gccatcgtga tggctagagt ttagccctcc cacacataac cagagggcag
     5401 caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta tggctttgat
     5461 cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca agatgcccct
     5521 gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag cagcagcagt
     5581 gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat tgacaccagt
     5641 gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt cagagatggg
     5701 gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt gagacaaagg
     5761 cttggccatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc
     5821 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     5881 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     5941 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     6001 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     6061 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     6121 ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc aagttcctct
     6181 ttcagaggtt atttcaggcc atggtgctgc gc
//
