LOCUS       sequence-423319-        7581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CCT5. C-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7581)
  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 7581)
  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_CCT5:1-541
FEATURES             Location/Qualifiers
     source          1..7581
                     /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             2756..3268
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3344..3362)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3399..3520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3410..3429)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3464..3483
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3765..4122
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3765..3956
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3788..3808)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3973..4108
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4153..5190
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5169..5177
                     /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    5214..5262
                     /note="synthetic polyadenylation signal"
     promoter        5418..5522
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5523..5591,5592..6383)
                     /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(5741..5760)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6541..7129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6621..6640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7245..7528
                     /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 caccatggcg tccatgggga ccctcgcctt cgatgaatat gggcgccctt tcctcatcat
     1141 caaggatcag gaccgcaagt cccgtcttat gggacttgag gccctcaagt ctcatataat
     1201 ggcagcaaag gctgtagcaa atacaatgag aacatcactt ggaccaaatg ggcttgataa
     1261 gatgatggtg gataaggatg gggatgtgac tgtaactaat gatggggcca ccatcttaag
     1321 catgatggat gttgatcatc agattgccaa gctgatggtg gaactgtcca agtctcagga
     1381 tgatgaaatt ggagatggaa ccacaggagt ggttgtcctg gctggtgcct tgttagaaga
     1441 agcggagcaa ttgctagacc gaggcattca cccaatcaga atagccgatg gctatgagca
     1501 ggctgctcgc gttgctattg aacacctgga caagatcagc gatagcgtcc ttgttgacat
     1561 aaaggacacc gaacccctga ttcagacagc aaaaaccacg ctgggctcca aagtggtcaa
     1621 cagttgtcac cgacagatgg ctgagattgc tgtgaatgcc gtcctcactg tagcagatat
     1681 ggagcggaga gacgttgact ttgagcttat caaagtagaa ggcaaagtgg gcggcaggct
     1741 ggaggacact aaactgatta agggcgtgat tgtggacaag gatttcagtc acccacagat
     1801 gccaaaaaaa gtggaagatg cgaagattgc aattctcaca tgtccatttg aaccacccaa
     1861 accaaaaaca aagcataagc tggatgtgac ctctgtcgaa gattataaag cccttcagaa
     1921 atacgaaaag gagaaatttg aagagatgat tcaacaaatt aaagagactg gtgctaacct
     1981 agcaatttgt cagtggggct ttgatgatga agcaaatcac ttacttcttc agaacaactt
     2041 gcctgcggtt cgctgggtag gaggacctga aattgagctg attgccatcg caacaggagg
     2101 gcggatcgtc cccaggttct cagagctcac agccgagaag ctgggctttg ctggtcttgt
     2161 acaggagatc tcatttggga caactaagga taaaatgctg gtcatcgagc agtgtaagaa
     2221 ctccagagct gtaaccattt ttattagagg aggaaataag atgatcattg aggaggcgaa
     2281 acgatccctt cacgatgctt tgtgtgtcat ccggaacctc atccgcgata atcgtgtggt
     2341 gtatggagga ggggctgctg agatatcctg tgccctggca gttagccaag aggcggataa
     2401 gtgccccacc ttagaacagt atgccatgag agcgtttgcc gacgcactgg aggtcatccc
     2461 catggccctc tctgaaaaca gtggcatgaa tcccatccag actatgaccg aagtccgagc
     2521 cagacaggtg aaggagatga accctgctct tggcatcgac tgtttgcaca aggggacaaa
     2581 tgatatgaag caacagcatg tcatagaaac cttgattggc aaaaagcaac agatatctct
     2641 tgcaacacaa atggttagaa tgattttgaa gattgatgac attcgtaagc ctggagaatc
     2701 tgaagaatct agatttgggc ccaattcctg caggcgagct ctcggctcga gcggcgtctt
     2761 cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt
     2821 ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat
     2881 ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc
     2941 gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta
     3001 ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg
     3061 ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga
     3121 cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg
     3181 cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg
     3241 gcggctgtgc gaacgcattc tggcgtaagg ccgcgactct agggcatgca agctgatccg
     3301 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     3361 gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3421 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3481 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3541 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     3601 caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     3661 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca
     3721 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     3781 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3841 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3901 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3961 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4021 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4081 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcga ttcttctgac
     4141 actagcgcca ccatgaagaa gcccgaactc accgctacca gcgttgaaaa atttctcatc
     4201 gagaagttcg acagtgtgag cgacctgatg cagttgtcgg agggcgaaga gagccgagcc
     4261 ttcagcttcg atgtcggcgg acgcggctat gtactgcggg tgaatagctg cgctgatggc
     4321 ttctacaaag accgctacgt gtaccgccac ttcgccagcg ctgcactacc catccccgaa
     4381 gtgttggaca tcggcgagtt cagcgagagc ctgacatact gcatcagtag acgcgcccaa
     4441 ggcgttactc tccaagacct ccccgaaaca gagctgcctg ctgtgttaca gcctgtcgcc
     4501 gaagctatgg atgctattgc cgccgccgac ctcagtcaaa ccagcggctt cggcccattc
     4561 gggccccaag gcatcggcca gtacacaacc tggcgggatt tcatttgcgc cattgctgat
     4621 ccccatgtct accactggca gaccgtgatg gacgacaccg tgtccgccag cgtagctcaa
     4681 gccctggacg aactgatgct gtgggccgaa gactgtcccg aggtgcgcca cctcgtccat
     4741 gccgacttcg gcagcaacaa cgtcctgacc gacaacggcc gcatcaccgc cgtaatcgac
     4801 tggtccgaag ctatgttcgg ggacagtcag tacgaggtgg ccaacatctt cttctggcgg
     4861 ccctggctgg cttgcatgga gcagcagact cgctacttcg agcgccggca tcccgagctg
     4921 gccggcagcc ctcgtctgcg agcctacatg ctgcgcatcg gcctggatca gctctaccag
     4981 agcctcgtgg acggcaactt cgacgatgct gcctgggctc aaggccgctg cgatgccatc
     5041 gtccgcagcg gggccggcac cgtcggtcgc acacaaatcg ctcgccggag cgcagccgta
     5101 tggaccgacg gctgcgtcga ggtgctggcc gacagcggca accgccggcc cagtacacga
     5161 ccgcgcgcta aggaggtagg tcgagtttag actctagcca tcacgatggc cgcaataaaa
     5221 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga
     5281 tcctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     5341 gtcagcaccg tttctgcgga ctggctttct acgtaatggt ttcttagacg tcaggtggca
     5401 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5461 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5521 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5581 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5641 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5701 ccgaagaacg ttttccaatg atgagcactt tcaaagttct gctatgtggc gcggtattat
     5761 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5821 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5881 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacta
     5941 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6001 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6061 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6121 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6181 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6241 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6301 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6361 cctcactgat taagcattgg taattcgaaa tgaccgacca agcgacgccc aaccggtatc
     6421 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6481 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6541 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6601 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6661 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6721 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6781 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6841 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6901 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6961 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7021 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7081 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt
     7141 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7201 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac
     7261 gcacgctgga tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga
     7321 ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat
     7381 ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct
     7441 gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc
     7501 acttactgtc tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc
     7561 cggcgtagag gatcgagatc t
//
