LOCUS       sequence-423360-        7138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CCNE1. N-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7138)
  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 7138)
  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_CCNE1:1-410
FEATURES             Location/Qualifiers
     source          1..7138
                     /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(2906..2924)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2956..3077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2967..2986)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3021..3040
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3322..3679
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3322..3513
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3345..3365)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3530..3665
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3710..4747
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4726..4734
                     /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    4771..4819
                     /note="synthetic polyadenylation signal"
     promoter        4975..5079
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5080..5148,5149..5940)
                     /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"
     rep_origin      complement(6098..6686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6178..6197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6802..7085
                     /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 ccgagggagc
     1621 gcagggagcg ggatgcgaag gagcgggaca ccatgaagga ggacggcggc gcggagttct
     1681 cggctcgctc caggaagagg aaggcaaacg tgaccgtttt tttgcaggat ccagatgaag
     1741 aaatggccaa aatcgacagg acggcgaggg accagtgtgg gagccagcct tgggacaata
     1801 atgcagtctg tgcagacccc tgctccctga tccccacacc tgacaaagaa gatgatgacc
     1861 gggtttaccc aaactcaacg tgcaagcctc ggattattgc accatccaga ggctccccgc
     1921 tgcctgtact gagctgggca aatagagagg aagtctggaa aatcatgtta aacaaggaaa
     1981 agacatactt aagggatcag cactttcttg agcaacaccc tcttctgcag ccaaaaatgc
     2041 gagcaattct tctggattgg ttaatggagg tgtgtgaagt ctataaactt cacagggaga
     2101 ccttttactt ggcacaagat ttctttgacc ggtatatggc gacacaagaa aatgttgtaa
     2161 aaactctttt acagcttatt gggatttcat ctttatttat tgcagccaaa cttgaggaaa
     2221 tctatcctcc aaagttgcac cagtttgcgt atgtgacaga tggagcttgt tcaggagatg
     2281 aaattctcac catggaatta atgattatga aggcccttaa gtggcgttta agtcccctga
     2341 ctattgtgtc ctggctgaat gtatacatgc aggttgcata tctaaatgac ttacatgaag
     2401 tgctactgcc gcagtatccc cagcaaatct ttatacagat tgcagagctg ttggatctct
     2461 gtgtcctgga tgttgactgc cttgaatttc cttatggtat acttgctgct tcggccttgt
     2521 atcatttctc gtcatctgaa ttgatgcaaa aggtttcagg gtatcagtgg tgcgacatag
     2581 agaactgtgt caagtggatg gttccatttg ccatggttat aagggagacg gggagctcaa
     2641 aactgaagca cttcaggggc gtcgctgatg aagatgcaca caacatacag acccacagag
     2701 acagcttgga tttgctggac aaagcccgag caaagaaagc catgttgtct gaacaaaata
     2761 gggcttctcc tctccccagt gggctcctca ccccgccaca gagcggtaag aagcagagca
     2821 gcgggccgga aatggcgtga gatatctaga tttgggccca attcctgcag gattttgcgg
     2881 ccgcttgctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggccgc
     2941 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3001 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3061 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3121 agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt
     3181 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     3241 gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag
     3301 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3361 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3421 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3481 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3541 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3601 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3661 aggcctaggc ttttgcaaaa agctcgattc ttctgacact agcgccacca tgaagaagcc
     3721 cgaactcacc gctaccagcg ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga
     3781 cctgatgcag ttgtcggagg gcgaagagag ccgagccttc agcttcgatg tcggcggacg
     3841 cggctatgta ctgcgggtga atagctgcgc tgatggcttc tacaaagacc gctacgtgta
     3901 ccgccacttc gccagcgctg cactacccat ccccgaagtg ttggacatcg gcgagttcag
     3961 cgagagcctg acatactgca tcagtagacg cgcccaaggc gttactctcc aagacctccc
     4021 cgaaacagag ctgcctgctg tgttacagcc tgtcgccgaa gctatggatg ctattgccgc
     4081 cgccgacctc agtcaaacca gcggcttcgg cccattcggg ccccaaggca tcggccagta
     4141 cacaacctgg cgggatttca tttgcgccat tgctgatccc catgtctacc actggcagac
     4201 cgtgatggac gacaccgtgt ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg
     4261 ggccgaagac tgtcccgagg tgcgccacct cgtccatgcc gacttcggca gcaacaacgt
     4321 cctgaccgac aacggccgca tcaccgccgt aatcgactgg tccgaagcta tgttcgggga
     4381 cagtcagtac gaggtggcca acatcttctt ctggcggccc tggctggctt gcatggagca
     4441 gcagactcgc tacttcgagc gccggcatcc cgagctggcc ggcagccctc gtctgcgagc
     4501 ctacatgctg cgcatcggcc tggatcagct ctaccagagc ctcgtggacg gcaacttcga
     4561 cgatgctgcc tgggctcaag gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt
     4621 cggtcgcaca caaatcgctc gccggagcgc agccgtatgg accgacggct gcgtcgaggt
     4681 gctggccgac agcggcaacc gccggcccag tacacgaccg cgcgctaagg aggtaggtcg
     4741 agtttagact ctagccatca cgatggccgc aataaaatat ctttattttc attacatctg
     4801 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc
     4861 cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg
     4921 gctttctacg taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4981 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5041 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5101 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5161 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5221 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5281 agcactttca aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5341 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5401 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5461 agtgataaca ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc
     5521 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5581 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5641 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5701 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5761 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5821 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5881 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5941 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     6001 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6061 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6121 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6181 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6241 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6301 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6361 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6421 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6481 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6541 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6601 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6661 ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg
     6721 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6781 tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg
     6841 ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg
     6901 taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt
     6961 ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt
     7021 ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc
     7081 atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
