LOCUS       sequence-423302-        7050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human DCAF1 fragment E1038 to E1400. 
            C-terminal NanoLuc luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7050)
  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 7050)
  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-C_DCAF1:1038-1400
FEATURES             Location/Qualifiers
     source          1..7050
                     /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"
     regulatory      1079..1088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2225..2737
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2813..2831)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2868..2989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2879..2898)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2933..2952
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3234..3591
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3234..3425
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3257..3277)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3442..3577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3622..4659
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4638..4646
                     /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    4683..4731
                     /note="synthetic polyadenylation signal"
     promoter        4887..4991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4992..5060,5061..5852)
                     /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(5210..5229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6010..6598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6090..6109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6714..6997
                     /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 ggctagcaaa gcgatcgctt ccgaattcgc
     1081 caccatggag ccaaaacaga ggcggcaagc gccaataaac tttacgtcaa ggctaaaccg
     1141 cagggcatca tttccaaagt atggaggggt ggatggcgga tgctttgata ggcaccttat
     1201 ctttagcaga ttccgtccta tttcagtgtt ccgggaagcc aatgaagatg agagtggctt
     1261 cacctgctgt gcattctcag cacgggagcg gttcctgatg cttggcacct gcacagggca
     1321 gctgaagctc tataatgtgt ttagtggaca ggaggaggcc agctataact gtcacaactc
     1381 agccatcaca catcttgaac cttccaggga tgggtccttg ctgctgacat ctgctacttg
     1441 gagccagcct ttgtctgcac tttggggaat gaagtcagta tttgatatga agcattcctt
     1501 cacagaagat cactatgttg agttcagtaa gcactcccag gatcgggtca tcggcacaaa
     1561 aggagacatt gcccacattt atgatattca gactggcaac aagctgttga ctctgtttaa
     1621 cccagatctt gccaacaact acaagaggaa ctgtgccacc tttaatccta cagatgatct
     1681 tgtcttaaat gatggcgtcc tctgggatgt ccgctctgca caggccatcc acaagtttga
     1741 caagttcaat atgaacatca gtggtgtttt ccatccaaat ggactggagg tgatcattaa
     1801 tactgagatt tgggaccttc gaacttttca tcttttgcat actgttcccg ctctggatca
     1861 gtgtcgcgtg gtgttcaatc acacgggaac agtgatgtat ggagctatgt tgcaggcaga
     1921 tgatgaagat gacttaatgg aagagaggat gaaaagcccc tttgggtcat ccttccgaac
     1981 atttaatgca actgactaca aacctatagc aaccattgat gtgaaacgga acatctttga
     2041 cctgtgtaca gacaccaaag actgctatct tgctgtcatt gagaatcaag gcagcatgga
     2101 tgccctgaac atggacacag tatgcaggct gtatgaagtg ggcaggcagc gtctggcaga
     2161 ggatgaggat gaagagtcta gatttgggcc caattcctgc aggcgagctc tcggctcgag
     2221 cggcgtcttc acactcgaag atttcgttgg ggactggcga cagacagccg gctacaacct
     2281 ggaccaagtc cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt
     2341 aactccgatc caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt
     2401 catcatcccg tatgaaggtc tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa
     2461 ggtggtgtac cctgtggatg atcatcactt taaggtgatc ctgcactatg gcacactggt
     2521 aatcgacggg gttacgccga acatgatcga ctatttcgga cggccgtatg aaggcatcgc
     2581 cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg tggaacggca acaaaattat
     2641 cgacgagcgc ctgatcaacc ccgacggctc cctgctgttc cgagtaacca tcaacggagt
     2701 gaccggctgg cggctgtgcg aacgcattct ggcgtaaggc cgcgactcta gggcatgcaa
     2761 gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     2821 caataactag cataacccct tggggcggcc gcttcgagca gacatgataa gatacattga
     2881 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2941 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3001 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     3061 aaacctctac aaatgtggta aaatcgaatt ttaacaaaat attaacgctt acaatttcct
     3121 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gcggatctgc
     3181 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     3241 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3301 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3361 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3421 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3481 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3541 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcgat
     3601 tcttctgaca ctagcgccac catgaagaag cccgaactca ccgctaccag cgttgaaaaa
     3661 tttctcatcg agaagttcga cagtgtgagc gacctgatgc agttgtcgga gggcgaagag
     3721 agccgagcct tcagcttcga tgtcggcgga cgcggctatg tactgcgggt gaatagctgc
     3781 gctgatggct tctacaaaga ccgctacgtg taccgccact tcgccagcgc tgcactaccc
     3841 atccccgaag tgttggacat cggcgagttc agcgagagcc tgacatactg catcagtaga
     3901 cgcgcccaag gcgttactct ccaagacctc cccgaaacag agctgcctgc tgtgttacag
     3961 cctgtcgccg aagctatgga tgctattgcc gccgccgacc tcagtcaaac cagcggcttc
     4021 ggcccattcg ggccccaagg catcggccag tacacaacct ggcgggattt catttgcgcc
     4081 attgctgatc cccatgtcta ccactggcag accgtgatgg acgacaccgt gtccgccagc
     4141 gtagctcaag ccctggacga actgatgctg tgggccgaag actgtcccga ggtgcgccac
     4201 ctcgtccatg ccgacttcgg cagcaacaac gtcctgaccg acaacggccg catcaccgcc
     4261 gtaatcgact ggtccgaagc tatgttcggg gacagtcagt acgaggtggc caacatcttc
     4321 ttctggcggc cctggctggc ttgcatggag cagcagactc gctacttcga gcgccggcat
     4381 cccgagctgg ccggcagccc tcgtctgcga gcctacatgc tgcgcatcgg cctggatcag
     4441 ctctaccaga gcctcgtgga cggcaacttc gacgatgctg cctgggctca aggccgctgc
     4501 gatgccatcg tccgcagcgg ggccggcacc gtcggtcgca cacaaatcgc tcgccggagc
     4561 gcagccgtat ggaccgacgg ctgcgtcgag gtgctggccg acagcggcaa ccgccggccc
     4621 agtacacgac cgcgcgctaa ggaggtaggt cgagtttaga ctctagccat cacgatggcc
     4681 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     4741 cgataaggat cctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat
     4801 tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtaatggtt tcttagacgt
     4861 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4921 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4981 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5041 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5101 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5161 gttttcgccc cgaagaacgt tttccaatga tgagcacttt caaagttctg ctatgtggcg
     5221 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5281 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5341 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5401 tgacaactat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5461 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5521 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5581 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5641 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5701 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5761 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5821 agataggtgc ctcactgatt aagcattggt aattcgaaat gaccgaccaa gcgacgccca
     5881 accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5941 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6001 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6061 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6181 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6421 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6541 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga
     6601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6661 gattttggtc atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa
     6721 tacaggaacg cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc
     6781 ttcagtggat taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac
     6841 ggttaaaatt tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac
     6901 ctgtctgctg ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat
     6961 ggtaaatcca cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca
     7021 cgatgcgtcc ggcgtagagg atcgagatct
//
