LOCUS       sequence-425944-        7695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CDC40. C-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7695)
  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 7695)
  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_CDC40:1-579
FEATURES             Location/Qualifiers
     source          1..7695
                     /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             2870..3382
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3458..3476)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3513..3634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3524..3543)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3578..3597
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3879..4236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3879..4070
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3902..3922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4087..4222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4267..5304
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5283..5291
                     /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    5328..5376
                     /note="synthetic polyadenylation signal"
     promoter        5532..5636
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5637..5705,5706..6497)
                     /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(5855..5874)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6655..7243)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6735..6754)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7359..7642
                     /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 caccatgtcg gctgcgattg cagctctggc cgcttcctat ggttcgggtt cagggtccga
     1141 atcggactcg gacagtgaga gcagtcggtg tccgctgcca gccgccgact ccctcatgca
     1201 cttgactaaa tcgccttcat caaagccgtc tctagcagtg gcagtggact cggctccgga
     1261 ggtggcagtt aaggaagatt tggagactgg agttcacctt gaccctgccg tcaaagaagt
     1321 tcagtataat cctacctatg agaccatgtt tgctcctgag tttggaccag aaaatccctt
     1381 taggacacag caaatggctg cccctagaaa tatgctttct ggatatgccg aaccagctca
     1441 tatcaatgat ttcatgtttg agcagcaaag gagaactttt gcaacatatg gttatgcatt
     1501 agacccttca ttagataatc atcaagtgtc tgctaaatat attggttctg tagaagaagc
     1561 tgaaaaaaat caaggtttaa ctgtatttga aactggtcag aagaaaacag aaaagaggaa
     1621 aaagtttaaa gaaaatgatg catccaatat tgatggtttt ttgggaccat gggcaaaata
     1681 tgtggatgaa aaagatgtag ccaaaccttc agaagaagag caaaaagaat tggatgaaat
     1741 cacagcaaag aggcagaaaa aaggaaaaca ggaagaagag aaacctgggg aggagaagac
     1801 aatcttacat gttaaagaaa tgtatgacta tcaaggcagg tcctatcttc acatacctca
     1861 ggatgttggt gttaatctac ggtcaactat gccacctgag aagtgttatc ttcccaaaaa
     1921 acaaattcat gtgtggtctg gacacacaaa gggcgtcagt gcagtcagat tgtttcctct
     1981 ctctggccat ttattgctgt cttgttccat ggactgtaaa attaagctat gggaggttta
     2041 tggagaacgg cgctgtctga gaacatttat tggtcacagt aaggctgtta gggatatctg
     2101 cttcaatact gcaggaacac agttcctcag tgcagcctat gacaggtatc ttaagctctg
     2161 ggacactgag acaggacagt gtatatcaag atttacaaac cgaaaagtac cttattgtgt
     2221 caaattcaat cctgatgaag ataagcaaaa tctctttgtg gctgggatgt ctgataagaa
     2281 gattgtgcaa tgggacattc gaagtggaga aattgtgcag gaatatgatc ggcatttggg
     2341 agctgtcaac accattgttt ttgtggatga gaataggaga tttgtgagca catctgatga
     2401 taaaagccta agagtttggg aatgggatat ccctgtggat ttcaagtaca tagcagaacc
     2461 cagtatgcac tcaatgcctg cagtgacttt gtctccaaat ggaaaatggc tagcatgcca
     2521 atcaatggac aaccaaatct taatttttgg agcacagaac agatttagat taaataagaa
     2581 aaaaattttt aagggccata tggtagcagg ctatgcttgt caggtggact tttcaccaga
     2641 catgagttat gtgatttcag gagatggaaa tggaaaatta aacatttggg actggaagac
     2701 cacaaaactc tacagtcgat ttaaagctca tgataaagtg tgtataggtg cagtgtggca
     2761 tcctcatgaa acttctaagg tcataacatg tggttgggat ggtctcatta aattgtggga
     2821 ttctagattt gggcccaatt cctgcaggcg agctctcggc tcgagcggcg tcttcacact
     2881 cgaagatttc gttggggact ggcgacagac agccggctac aacctggacc aagtccttga
     2941 acagggaggt gtgtccagtt tgtttcagaa tctcggggtg tccgtaactc cgatccaaag
     3001 gattgtcctg agcggtgaaa atgggctgaa gatcgacatc catgtcatca tcccgtatga
     3061 aggtctgagc ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg tgtaccctgt
     3121 ggatgatcat cactttaagg tgatcctgca ctatggcaca ctggtaatcg acggggttac
     3181 gccgaacatg atcgactatt tcggacggcc gtatgaaggc atcgccgtgt tcgacggcaa
     3241 aaagatcact gtaacaggga ccctgtggaa cggcaacaaa attatcgacg agcgcctgat
     3301 caaccccgac ggctccctgc tgttccgagt aaccatcaac ggagtgaccg gctggcggct
     3361 gtgcgaacgc attctggcgt aaggccgcga ctctagggca tgcaagctga tccggctgct
     3421 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     3481 ccccttgggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     3541 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3601 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3661 gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg
     3721 tggtaaaatc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     3781 ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc accatggcct
     3841 gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg
     3901 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3961 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4021 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4081 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4141 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4201 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcgattcttc tgacactagc
     4261 gccaccatga agaagcccga actcaccgct accagcgttg aaaaatttct catcgagaag
     4321 ttcgacagtg tgagcgacct gatgcagttg tcggagggcg aagagagccg agccttcagc
     4381 ttcgatgtcg gcggacgcgg ctatgtactg cgggtgaata gctgcgctga tggcttctac
     4441 aaagaccgct acgtgtaccg ccacttcgcc agcgctgcac tacccatccc cgaagtgttg
     4501 gacatcggcg agttcagcga gagcctgaca tactgcatca gtagacgcgc ccaaggcgtt
     4561 actctccaag acctccccga aacagagctg cctgctgtgt tacagcctgt cgccgaagct
     4621 atggatgcta ttgccgccgc cgacctcagt caaaccagcg gcttcggccc attcgggccc
     4681 caaggcatcg gccagtacac aacctggcgg gatttcattt gcgccattgc tgatccccat
     4741 gtctaccact ggcagaccgt gatggacgac accgtgtccg ccagcgtagc tcaagccctg
     4801 gacgaactga tgctgtgggc cgaagactgt cccgaggtgc gccacctcgt ccatgccgac
     4861 ttcggcagca acaacgtcct gaccgacaac ggccgcatca ccgccgtaat cgactggtcc
     4921 gaagctatgt tcggggacag tcagtacgag gtggccaaca tcttcttctg gcggccctgg
     4981 ctggcttgca tggagcagca gactcgctac ttcgagcgcc ggcatcccga gctggccggc
     5041 agccctcgtc tgcgagccta catgctgcgc atcggcctgg atcagctcta ccagagcctc
     5101 gtggacggca acttcgacga tgctgcctgg gctcaaggcc gctgcgatgc catcgtccgc
     5161 agcggggccg gcaccgtcgg tcgcacacaa atcgctcgcc ggagcgcagc cgtatggacc
     5221 gacggctgcg tcgaggtgct ggccgacagc ggcaaccgcc ggcccagtac acgaccgcgc
     5281 gctaaggagg taggtcgagt ttagactcta gccatcacga tggccgcaat aaaatatctt
     5341 tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagcgata aggatcctct
     5401 ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc
     5461 accgtttctg cggactggct ttctacgtaa tggtttctta gacgtcaggt ggcacttttc
     5521 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5581 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5641 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5701 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5761 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5821 aacgttttcc aatgatgagc actttcaaag ttctgctatg tggcgcggta ttatcccgta
     5881 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5941 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6001 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca actatcggag
     6061 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6121 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6181 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6241 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6301 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6361 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6421 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6481 tgattaagca ttggtaattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca
     6541 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6601 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6661 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6721 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6781 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6841 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6901 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6961 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7021 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7081 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7141 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7201 tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt
     7261 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7321 attatcaaaa aggatcttca cctagatcct tttatagtcc ggaaatacag gaacgcacgc
     7381 tggatggccc ttcgctggga tggtgaaacc atgaaaaatg gcagcttcag tggattaagt
     7441 gggggtaatg tggcctgtac cctctggttg cataggtatt catacggtta aaatttatca
     7501 ggcgcgattg cggcagtttt tcgggtggtt tgttgccatt tttacctgtc tgctgccgtg
     7561 atcgcgctga acgcgtttta gcggtgcgta caattaaggg attatggtaa atccacttac
     7621 tgtctgccct cgtagccatc gagataaacc gcagtactcc ggccacgatg cgtccggcgt
     7681 agaggatcga gatct
//
