LOCUS       sequence-423318-        7530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CCT5. N-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7530)
  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 7530)
  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_CCT5:1-541
FEATURES             Location/Qualifiers
     source          1..7530
                     /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(3298..3316)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3348..3469
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3359..3378)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3413..3432
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3714..4071
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3714..3905
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3737..3757)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3922..4057
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4102..5139
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5118..5126
                     /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    5163..5211
                     /note="synthetic polyadenylation signal"
     promoter        5367..5471
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5472..5540,5541..6332)
                     /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(5690..5709)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6490..7078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6570..6589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7194..7477
                     /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 gcgtccatgg
     1621 ggaccctcgc cttcgatgaa tatgggcgcc ctttcctcat catcaaggat caggaccgca
     1681 agtcccgtct tatgggactt gaggccctca agtctcatat aatggcagca aaggctgtag
     1741 caaatacaat gagaacatca cttggaccaa atgggcttga taagatgatg gtggataagg
     1801 atggggatgt gactgtaact aatgatgggg ccaccatctt aagcatgatg gatgttgatc
     1861 atcagattgc caagctgatg gtggaactgt ccaagtctca ggatgatgaa attggagatg
     1921 gaaccacagg agtggttgtc ctggctggtg ccttgttaga agaagcggag caattgctag
     1981 accgaggcat tcacccaatc agaatagccg atggctatga gcaggctgct cgcgttgcta
     2041 ttgaacacct ggacaagatc agcgatagcg tccttgttga cataaaggac accgaacccc
     2101 tgattcagac agcaaaaacc acgctgggct ccaaagtggt caacagttgt caccgacaga
     2161 tggctgagat tgctgtgaat gccgtcctca ctgtagcaga tatggagcgg agagacgttg
     2221 actttgagct tatcaaagta gaaggcaaag tgggcggcag gctggaggac actaaactga
     2281 ttaagggcgt gattgtggac aaggatttca gtcacccaca gatgccaaaa aaagtggaag
     2341 atgcgaagat tgcaattctc acatgtccat ttgaaccacc caaaccaaaa acaaagcata
     2401 agctggatgt gacctctgtc gaagattata aagcccttca gaaatacgaa aaggagaaat
     2461 ttgaagagat gattcaacaa attaaagaga ctggtgctaa cctagcaatt tgtcagtggg
     2521 gctttgatga tgaagcaaat cacttacttc ttcagaacaa cttgcctgcg gttcgctggg
     2581 taggaggacc tgaaattgag ctgattgcca tcgcaacagg agggcggatc gtccccaggt
     2641 tctcagagct cacagccgag aagctgggct ttgctggtct tgtacaggag atctcatttg
     2701 ggacaactaa ggataaaatg ctggtcatcg agcagtgtaa gaactccaga gctgtaacca
     2761 tttttattag aggaggaaat aagatgatca ttgaggaggc gaaacgatcc cttcacgatg
     2821 ctttgtgtgt catccggaac ctcatccgcg ataatcgtgt ggtgtatgga ggaggggctg
     2881 ctgagatatc ctgtgccctg gcagttagcc aagaggcgga taagtgcccc accttagaac
     2941 agtatgccat gagagcgttt gccgacgcac tggaggtcat ccccatggcc ctctctgaaa
     3001 acagtggcat gaatcccatc cagactatga ccgaagtccg agccagacag gtgaaggaga
     3061 tgaaccctgc tcttggcatc gactgtttgc acaaggggac aaatgatatg aagcaacagc
     3121 atgtcataga aaccttgatt ggcaaaaagc aacagatatc tcttgcaaca caaatggtta
     3181 gaatgatttt gaagattgat gacattcgta agcctggaga atctgaagaa taataatcta
     3241 gatttgggcc caattcctgc aggattttgc ggccgcttgc tgagttggct gctgccaccg
     3301 ctgagcaata actagcataa ccccttggcc gcttcgagca gacatgataa gatacattga
     3361 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3421 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3481 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     3541 aaacctctac aaatgtggta aaatcgaatt ttaacaaaat attaacgctt acaatttcct
     3601 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gcggatctgc
     3661 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     3721 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3781 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3841 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3901 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3961 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4021 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcgat
     4081 tcttctgaca ctagcgccac catgaagaag cccgaactca ccgctaccag cgttgaaaaa
     4141 tttctcatcg agaagttcga cagtgtgagc gacctgatgc agttgtcgga gggcgaagag
     4201 agccgagcct tcagcttcga tgtcggcgga cgcggctatg tactgcgggt gaatagctgc
     4261 gctgatggct tctacaaaga ccgctacgtg taccgccact tcgccagcgc tgcactaccc
     4321 atccccgaag tgttggacat cggcgagttc agcgagagcc tgacatactg catcagtaga
     4381 cgcgcccaag gcgttactct ccaagacctc cccgaaacag agctgcctgc tgtgttacag
     4441 cctgtcgccg aagctatgga tgctattgcc gccgccgacc tcagtcaaac cagcggcttc
     4501 ggcccattcg ggccccaagg catcggccag tacacaacct ggcgggattt catttgcgcc
     4561 attgctgatc cccatgtcta ccactggcag accgtgatgg acgacaccgt gtccgccagc
     4621 gtagctcaag ccctggacga actgatgctg tgggccgaag actgtcccga ggtgcgccac
     4681 ctcgtccatg ccgacttcgg cagcaacaac gtcctgaccg acaacggccg catcaccgcc
     4741 gtaatcgact ggtccgaagc tatgttcggg gacagtcagt acgaggtggc caacatcttc
     4801 ttctggcggc cctggctggc ttgcatggag cagcagactc gctacttcga gcgccggcat
     4861 cccgagctgg ccggcagccc tcgtctgcga gcctacatgc tgcgcatcgg cctggatcag
     4921 ctctaccaga gcctcgtgga cggcaacttc gacgatgctg cctgggctca aggccgctgc
     4981 gatgccatcg tccgcagcgg ggccggcacc gtcggtcgca cacaaatcgc tcgccggagc
     5041 gcagccgtat ggaccgacgg ctgcgtcgag gtgctggccg acagcggcaa ccgccggccc
     5101 agtacacgac cgcgcgctaa ggaggtaggt cgagtttaga ctctagccat cacgatggcc
     5161 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     5221 cgataaggat cctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat
     5281 tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtaatggtt tcttagacgt
     5341 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5401 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5461 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5521 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5581 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5641 gttttcgccc cgaagaacgt tttccaatga tgagcacttt caaagttctg ctatgtggcg
     5701 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5761 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5821 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5881 tgacaactat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5941 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6001 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6061 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6121 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6181 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6241 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6301 agataggtgc ctcactgatt aagcattggt aattcgaaat gaccgaccaa gcgacgccca
     6361 accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6421 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6481 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6541 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6601 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6661 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6721 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6781 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6841 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6901 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6961 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7021 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga
     7081 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7141 gattttggtc atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa
     7201 tacaggaacg cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc
     7261 ttcagtggat taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac
     7321 ggttaaaatt tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac
     7381 ctgtctgctg ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat
     7441 ggtaaatcca cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca
     7501 cgatgcgtcc ggcgtagagg atcgagatct
//
