LOCUS       sequence-425947-        7188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DDB2. N-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7188)
  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 7188)
  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-N_DDB2:1-427
FEATURES             Location/Qualifiers
     source          1..7188
                     /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"
     CDS             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(2956..2974)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3006..3127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3017..3036)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3071..3090
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3372..3729
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3372..3563
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3395..3415)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3580..3715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3760..4797
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4776..4784
                     /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    4821..4869
                     /note="synthetic polyadenylation signal"
     promoter        5025..5129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5130..5198,5199..5990)
                     /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(5348..5367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6148..6736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6228..6247)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6852..7135
                     /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 ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgcttc cgaattcatg gctcccaaga
     1621 aacgcccaga aacccagaag acctccgaga ttgtattacg ccccaggaac aagaggagca
     1681 ggagtcccct ggagctggag cccgaggcca agaagctctg tgcgaagggc tccggtccta
     1741 gcagaagatg tgactcagac tgcctctggg tggggctggc tggcccacag atcctgccac
     1801 catgccgcag catcgtcagg accctccacc agcataagct gggcagagct tcctggccat
     1861 ctgtccagca ggggctccag cagtcctttt tgcacactct ggattcttac cggatattac
     1921 aaaaggctgc cccctttgac aggagggcta catccttggc gtggcaccca actcacccca
     1981 gcaccgtggc tgtgggttcc aaagggggag atatcatgct ctggaatttt ggcatcaagg
     2041 acaaacccac cttcatcaaa gggattggag ctggagggag catcactggg ctgaagttta
     2101 accctctcaa taccaaccag ttttacgcct cctcaatgga gggaacaact aggctgcaag
     2161 actttaaagg caacattcta cgagtttttg ccagctcaga caccatcaac atctggtttt
     2221 gtagcctgga tgtgtctgct agtagccgaa tggtggtcac aggagacaac gtggggaacg
     2281 tgatcctgct gaacatggac ggcaaagagc tttggaatct cagaatgcac aaaaagaaag
     2341 tgacgcatgt ggccctgaac ccatgctgtg attggttcct ggccacagcc tccgtagatc
     2401 aaacagtgaa aatttgggac ctgcgccagg ttagagggaa agccagcttc ctctactcgc
     2461 tgccgcacag gcatcctgtc aacgcagctt gtttcagtcc cgatggagcc cggctcctga
     2521 ccacggacca gaagagcgag atccgagttt actctgcttc ccagtgggac tgccccctgg
     2581 gcctgatccc gcaccctcac cgtcacttcc agcacctcac acctatcaag gcagcctggc
     2641 atcctcgcta caacctcatt gttgtgggcc gatacccaga tcctaatttc aaaagttgta
     2701 ccccttatga attgaggacg atcgacgtgt tcgatggaaa ctcagggaag atgatgtgtc
     2761 agctctatga cccagaatct tctggcatca gttcgcttaa tgaattcaat cccatggggg
     2821 acacgctggc ctctgcaatg ggttaccaca ttctcatctg gagccaggag gaagccagga
     2881 cacggaagta ataatctaga tttgggccca attcctgcag gattttgcgg ccgcttgctg
     2941 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggccgc ttcgagcaga
     3001 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3061 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3121 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     3181 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat
     3241 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3301 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     3361 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3421 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3481 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3541 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3601 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3661 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3721 ttttgcaaaa agctcgattc ttctgacact agcgccacca tgaagaagcc cgaactcacc
     3781 gctaccagcg ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga cctgatgcag
     3841 ttgtcggagg gcgaagagag ccgagccttc agcttcgatg tcggcggacg cggctatgta
     3901 ctgcgggtga atagctgcgc tgatggcttc tacaaagacc gctacgtgta ccgccacttc
     3961 gccagcgctg cactacccat ccccgaagtg ttggacatcg gcgagttcag cgagagcctg
     4021 acatactgca tcagtagacg cgcccaaggc gttactctcc aagacctccc cgaaacagag
     4081 ctgcctgctg tgttacagcc tgtcgccgaa gctatggatg ctattgccgc cgccgacctc
     4141 agtcaaacca gcggcttcgg cccattcggg ccccaaggca tcggccagta cacaacctgg
     4201 cgggatttca tttgcgccat tgctgatccc catgtctacc actggcagac cgtgatggac
     4261 gacaccgtgt ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg ggccgaagac
     4321 tgtcccgagg tgcgccacct cgtccatgcc gacttcggca gcaacaacgt cctgaccgac
     4381 aacggccgca tcaccgccgt aatcgactgg tccgaagcta tgttcgggga cagtcagtac
     4441 gaggtggcca acatcttctt ctggcggccc tggctggctt gcatggagca gcagactcgc
     4501 tacttcgagc gccggcatcc cgagctggcc ggcagccctc gtctgcgagc ctacatgctg
     4561 cgcatcggcc tggatcagct ctaccagagc ctcgtggacg gcaacttcga cgatgctgcc
     4621 tgggctcaag gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt cggtcgcaca
     4681 caaatcgctc gccggagcgc agccgtatgg accgacggct gcgtcgaggt gctggccgac
     4741 agcggcaacc gccggcccag tacacgaccg cgcgctaagg aggtaggtcg agtttagact
     4801 ctagccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     4861 ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga
     4921 tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg
     4981 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5041 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5101 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5161 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5221 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5281 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttca
     5341 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5401 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5461 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5521 ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc gcttttttgc
     5581 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5641 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5701 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5761 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5821 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5881 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5941 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ttcgaaatga
     6001 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6061 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6121 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6181 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6241 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6301 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6361 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6421 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6481 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6541 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6601 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6661 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6721 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6781 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6841 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     6901 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     6961 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     7021 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     7081 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     7141 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
