LOCUS       sequence-425993-        7404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CDC20. N-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7404)
  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 7404)
  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_CDC20:1-499
FEATURES             Location/Qualifiers
     source          1..7404
                     /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(3172..3190)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3222..3343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3233..3252)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3287..3306
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3588..3945
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3588..3779
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3611..3631)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3796..3931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3976..5013
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4992..5000
                     /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    5037..5085
                     /note="synthetic polyadenylation signal"
     promoter        5241..5345
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5346..5414,5415..6206)
                     /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(5564..5583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6364..6952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6444..6463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7068..7351
                     /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 gcacagttcg
     1621 cgttcgagag tgacctgcac tcgctgcttc agctggatgc acccatcccc aatgcacccc
     1681 ctgcgcgctg gcagcgcaaa gccaaggaag ccgcaggccc ggccccctca cccatgcggg
     1741 ccgccaaccg atcccacagc gccggcagga ctccgggccg aactcctggc aaatccagtt
     1801 ccaaggttca gaccactcct agcaaacctg gcggtgaccg ctatatcccc catcgcagtg
     1861 ctgcccagat ggaggtggcc agcttcctcc tgagcaagga gaaccagcct gaaaacagcc
     1921 agacgcccac caagaaggaa catcagaaag cctgggcttt gaacctgaac ggttttgatg
     1981 tagaggaagc caagatcctt cggctcagtg gaaaaccaca aaatgcgcca gagggttacc
     2041 agaacagact gaaagtactc tacagccaaa aggccactcc tggctccagc cggaagacct
     2101 gccgttacat tccttccctg ccagaccgta tcctggatgc gcctgaaatc cgaaatgact
     2161 attacctgaa ccttgtggat tggagttctg ggaatgtact ggccgtggca ctggacaaca
     2221 gtgtgtacct gtggagtgca agctctggtg acatcctgca gcttttgcaa atggagcagc
     2281 ctggggaata tatatcctct gtggcctgga tcaaagaggg caactacttg gctgtgggca
     2341 ccagcagtgc tgaggtgcag ctatgggatg tgcagcagca gaaacggctt cgaaatatga
     2401 ccagtcactc tgcccgagtg ggctccctaa gctggaacag ctatatcctg tccagtggtt
     2461 cacgttctgg ccacatccac caccatgatg ttcgggtagc agaacaccat gtggccacac
     2521 tgagtggcca cagccaggaa gtgtgtgggc tgcgctgggc cccagatgga cgacatttgg
     2581 ccagtggtgg taatgataac ttggtcaatg tgtggcctag tgctcctgga gagggtggct
     2641 gggttcctct gcagacattc acccagcatc aaggggctgt caaggccgta gcatggtgtc
     2701 cctggcagtc caatgtcctg gcaacaggag ggggcaccag tgatcgacac attcgcatct
     2761 ggaatgtgtg ctctggggcc tgtctgagtg ccgtggatgc ccattcccag gtgtgctcca
     2821 tcctctggtc tccccattac aaggagctca tctcaggcca tggctttgca cagaaccagc
     2881 tagttatttg gaagtaccca accatggcca aggtggctga actcaaaggt cacacatccc
     2941 gggtcctgag tctgaccatg agcccagatg gggccacagt ggcatccgca gcagcagatg
     3001 agaccctgag gctatggcgc tgttttgagt tggaccctgc gcggcggcgg gagcgggaga
     3061 aggccagtgc agccaaaagc agcctcatcc accaaggcat ccgctaataa tctagatttg
     3121 ggcccaattc ctgcaggatt ttgcggccgc ttgctgagtt ggctgctgcc accgctgagc
     3181 aataactagc ataacccctt ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3241 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3301 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3361 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     3421 ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     3481 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca
     3541 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     3601 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3781 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3841 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3901 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cgattcttct
     3961 gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta ccagcgttga aaaatttctc
     4021 atcgagaagt tcgacagtgt gagcgacctg atgcagttgt cggagggcga agagagccga
     4081 gccttcagct tcgatgtcgg cggacgcggc tatgtactgc gggtgaatag ctgcgctgat
     4141 ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca gcgctgcact acccatcccc
     4201 gaagtgttgg acatcggcga gttcagcgag agcctgacat actgcatcag tagacgcgcc
     4261 caaggcgtta ctctccaaga cctccccgaa acagagctgc ctgctgtgtt acagcctgtc
     4321 gccgaagcta tggatgctat tgccgccgcc gacctcagtc aaaccagcgg cttcggccca
     4381 ttcgggcccc aaggcatcgg ccagtacaca acctggcggg atttcatttg cgccattgct
     4441 gatccccatg tctaccactg gcagaccgtg atggacgaca ccgtgtccgc cagcgtagct
     4501 caagccctgg acgaactgat gctgtgggcc gaagactgtc ccgaggtgcg ccacctcgtc
     4561 catgccgact tcggcagcaa caacgtcctg accgacaacg gccgcatcac cgccgtaatc
     4621 gactggtccg aagctatgtt cggggacagt cagtacgagg tggccaacat cttcttctgg
     4681 cggccctggc tggcttgcat ggagcagcag actcgctact tcgagcgccg gcatcccgag
     4741 ctggccggca gccctcgtct gcgagcctac atgctgcgca tcggcctgga tcagctctac
     4801 cagagcctcg tggacggcaa cttcgacgat gctgcctggg ctcaaggccg ctgcgatgcc
     4861 atcgtccgca gcggggccgg caccgtcggt cgcacacaaa tcgctcgccg gagcgcagcc
     4921 gtatggaccg acggctgcgt cgaggtgctg gccgacagcg gcaaccgccg gcccagtaca
     4981 cgaccgcgcg ctaaggaggt aggtcgagtt tagactctag ccatcacgat ggccgcaata
     5041 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa
     5101 ggatcctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc
     5161 ggggtcagca ccgtttctgc ggactggctt tctacgtaat ggtttcttag acgtcaggtg
     5221 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5281 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5341 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5401 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5461 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5521 gccccgaaga acgttttcca atgatgagca ctttcaaagt tctgctatgt ggcgcggtat
     5581 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5641 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5701 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5761 ctatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5821 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5881 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5941 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6001 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6061 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6121 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6181 gtgcctcact gattaagcat tggtaattcg aaatgaccga ccaagcgacg cccaaccggt
     6241 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6301 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6361 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6421 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6481 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6541 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6601 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6661 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6721 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6781 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6841 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6901 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     6961 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7021 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg
     7081 aacgcacgct ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt
     7141 ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa
     7201 aatttatcag gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct
     7261 gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa
     7321 tccacttact gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc
     7381 gtccggcgta gaggatcgag atct
//
