LOCUS       sequence-425959-        7008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CSNK2A2. C-terminal 
            NanoLuc luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7008)
  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 7008)
  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_CSNK2A2:1-350
FEATURES             Location/Qualifiers
     source          1..7008
                     /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             2183..2695
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2771..2789)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2826..2947
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2837..2856)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2891..2910
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3192..3549
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3192..3383
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3215..3235)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3400..3535
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3580..4617
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4596..4604
                     /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    4641..4689
                     /note="synthetic polyadenylation signal"
     promoter        4845..4949
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4950..5018,5019..5810)
                     /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(5168..5187)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5968..6556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6048..6067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6672..6955
                     /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 caccatgccc ggcccggccg cgggcagcag ggcccgggtc tacgccgagg tgaacagtct
     1141 gaggagccgc gagtactggg actacgaggc tcacgtcccg agctggggta atcaagatga
     1201 ttaccaactg gttcgaaaac ttggtcgggg aaaatatagt gaagtatttg aggccattaa
     1261 tatcaccaac aatgagagag tggttgtaaa aatcctgaag ccagtgaaga aaaagaagat
     1321 aaaacgagag gttaagattc tggagaacct tcgtggtgga acaaatatca ttaagctgat
     1381 tgacactgta aaggaccccg tgtcaaagac accagctttg gtatttgaat atatcaataa
     1441 tacagatttt aagcaactct accagatcct gacagacttt gatatccggt tttatatgta
     1501 tgaactactt aaagctctgg attactgcca cagcaaggga atcatgcaca gggatgtgaa
     1561 acctcacaat gtcatgatag atcaccaaca gaaaaagctg cgactgatag attggggtct
     1621 ggcagaattc tatcatcctg ctcaggagta caatgttcgt gtagcctcaa ggtacttcaa
     1681 gggaccagag ctcctcgtgg actatcagat gtatgattat agcttggaca tgtggagttt
     1741 gggctgtatg ttagcaagca tgatctttcg aagggaacca ttcttccatg gacaggacaa
     1801 ctatgaccag cttgttcgca ttgccaaggt tctgggtaca gaagaactgt atgggtatct
     1861 gaagaagtat cacatagacc tagatccaca cttcaacgat atcctgggac aacattcacg
     1921 gaaacgctgg gaaaacttta tccatagtga gaacagacac cttgtcagcc ctgaggccct
     1981 agatcttctg gacaaacttc tgcgatacga ccatcaacag agactgactg ccaaagaggc
     2041 catggagcac ccatacttct accctgtggt gaaggagcag tcccagcctt gtgcagacaa
     2101 tgctgtgctt tccagtggtc tcacggcagc acgatctaga tttgggccca attcctgcag
     2161 gcgagctctc ggctcgagcg gcgtcttcac actcgaagat ttcgttgggg actggcgaca
     2221 gacagccggc tacaacctgg accaagtcct tgaacaggga ggtgtgtcca gtttgtttca
     2281 gaatctcggg gtgtccgtaa ctccgatcca aaggattgtc ctgagcggtg aaaatgggct
     2341 gaagatcgac atccatgtca tcatcccgta tgaaggtctg agcggcgacc aaatgggcca
     2401 gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat catcacttta aggtgatcct
     2461 gcactatggc acactggtaa tcgacggggt tacgccgaac atgatcgact atttcggacg
     2521 gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc actgtaacag ggaccctgtg
     2581 gaacggcaac aaaattatcg acgagcgcct gatcaacccc gacggctccc tgctgttccg
     2641 agtaaccatc aacggagtga ccggctggcg gctgtgcgaa cgcattctgg cgtaaggccg
     2701 cgactctagg gcatgcaagc tgatccggct gctaacaaag cccgaaagga agctgagttg
     2761 gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc ttcgagcaga
     2821 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2881 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2941 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     3001 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat
     3061 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3121 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     3181 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3241 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3301 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3361 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3421 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3481 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3541 ttttgcaaaa agctcgattc ttctgacact agcgccacca tgaagaagcc cgaactcacc
     3601 gctaccagcg ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga cctgatgcag
     3661 ttgtcggagg gcgaagagag ccgagccttc agcttcgatg tcggcggacg cggctatgta
     3721 ctgcgggtga atagctgcgc tgatggcttc tacaaagacc gctacgtgta ccgccacttc
     3781 gccagcgctg cactacccat ccccgaagtg ttggacatcg gcgagttcag cgagagcctg
     3841 acatactgca tcagtagacg cgcccaaggc gttactctcc aagacctccc cgaaacagag
     3901 ctgcctgctg tgttacagcc tgtcgccgaa gctatggatg ctattgccgc cgccgacctc
     3961 agtcaaacca gcggcttcgg cccattcggg ccccaaggca tcggccagta cacaacctgg
     4021 cgggatttca tttgcgccat tgctgatccc catgtctacc actggcagac cgtgatggac
     4081 gacaccgtgt ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg ggccgaagac
     4141 tgtcccgagg tgcgccacct cgtccatgcc gacttcggca gcaacaacgt cctgaccgac
     4201 aacggccgca tcaccgccgt aatcgactgg tccgaagcta tgttcgggga cagtcagtac
     4261 gaggtggcca acatcttctt ctggcggccc tggctggctt gcatggagca gcagactcgc
     4321 tacttcgagc gccggcatcc cgagctggcc ggcagccctc gtctgcgagc ctacatgctg
     4381 cgcatcggcc tggatcagct ctaccagagc ctcgtggacg gcaacttcga cgatgctgcc
     4441 tgggctcaag gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt cggtcgcaca
     4501 caaatcgctc gccggagcgc agccgtatgg accgacggct gcgtcgaggt gctggccgac
     4561 agcggcaacc gccggcccag tacacgaccg cgcgctaagg aggtaggtcg agtttagact
     4621 ctagccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     4681 ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga
     4741 tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg
     4801 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4861 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4921 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4981 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5041 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5101 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttca
     5161 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5221 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5281 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5341 ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc gcttttttgc
     5401 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5461 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5521 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5581 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5641 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5701 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5761 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ttcgaaatga
     5821 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5881 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5941 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6001 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6061 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6121 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6181 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6241 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6301 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6361 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6421 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6481 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6541 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6601 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6661 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     6721 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     6781 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     6841 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     6901 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     6961 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
