LOCUS       sequence-423386-        6021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CDC40. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6021)
  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 6021)
  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: pBiT3.1-N_CDC40:1-579
FEATURES             Location/Qualifiers
     source          1..6021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3084..3205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3095..3114)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3320..3353)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3363..3410
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3442..4236
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3496..3515)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4106..4125
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4394..4982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4474..4493)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5108..5156
                     /note="synthetic polyadenylation signal"
     CDS             complement(5180..5578)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5575..5584
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5609..5966)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5623..5758
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5775..5966
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5923..5943
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggtta gcgatcgcca tggtgagcgg
     1201 ctggcggctg ttcaagaaga ttagcgggag ctccggtggc tcgagcggta tgtcggctgc
     1261 gattgcagct ctggccgctt cctatggttc gggttcaggg tccgaatcgg actcggacag
     1321 tgagagcagt cggtgtccgc tgccagccgc cgactccctc atgcacttga ctaaatcgcc
     1381 ttcatcaaag ccgtctctag cagtggcagt ggactcggct ccggaggtgg cagttaagga
     1441 agatttggag actggagttc accttgaccc tgccgtcaaa gaagttcagt ataatcctac
     1501 ctatgagacc atgtttgctc ctgagtttgg accagaaaat ccctttagga cacagcaaat
     1561 ggctgcccct agaaatatgc tttctggata tgccgaacca gctcatatca atgatttcat
     1621 gtttgagcag caaaggagaa cttttgcaac atatggttat gcattagacc cttcattaga
     1681 taatcatcaa gtgtctgcta aatatattgg ttctgtagaa gaagctgaaa aaaatcaagg
     1741 tttaactgta tttgaaactg gtcagaagaa aacagaaaag aggaaaaagt ttaaagaaaa
     1801 tgatgcatcc aatattgatg gttttttggg accatgggca aaatatgtgg atgaaaaaga
     1861 tgtagccaaa ccttcagaag aagagcaaaa agaattggat gaaatcacag caaagaggca
     1921 gaaaaaagga aaacaggaag aagagaaacc tggggaggag aagacaatct tacatgttaa
     1981 agaaatgtat gactatcaag gcaggtccta tcttcacata cctcaggatg ttggtgttaa
     2041 tctacggtca actatgccac ctgagaagtg ttatcttccc aaaaaacaaa ttcatgtgtg
     2101 gtctggacac acaaagggcg tcagtgcagt cagattgttt cctctctctg gccatttatt
     2161 gctgtcttgt tccatggact gtaaaattaa gctatgggag gtttatggag aacggcgctg
     2221 tctgagaaca tttattggtc acagtaaggc tgttagggat atctgcttca atactgcagg
     2281 aacacagttc ctcagtgcag cctatgacag gtatcttaag ctctgggaca ctgagacagg
     2341 acagtgtata tcaagattta caaaccgaaa agtaccttat tgtgtcaaat tcaatcctga
     2401 tgaagataag caaaatctct ttgtggctgg gatgtctgat aagaagattg tgcaatggga
     2461 cattcgaagt ggagaaattg tgcaggaata tgatcggcat ttgggagctg tcaacaccat
     2521 tgtttttgtg gatgagaata ggagatttgt gagcacatct gatgataaaa gcctaagagt
     2581 ttgggaatgg gatatccctg tggatttcaa gtacatagca gaacccagta tgcactcaat
     2641 gcctgcagtg actttgtctc caaatggaaa atggctagca tgccaatcaa tggacaacca
     2701 aatcttaatt tttggagcac agaacagatt tagattaaat aagaaaaaaa tttttaaggg
     2761 ccatatggta gcaggctatg cttgtcaggt ggacttttca ccagacatga gttatgtgat
     2821 ttcaggagat ggaaatggaa aattaaacat ttgggactgg aagaccacaa aactctacag
     2881 tcgatttaaa gctcatgata aagtgtgtat aggtgcagtg tggcatcctc atgaaacttc
     2941 taaggtcata acatgtggtt gggatggtct cattaaattg tgggattaat ctagaggtca
     3001 agcttcgggg gtttaaacct gcaggcatgc aagctgattt ggctgctgcc accgctgagc
     3061 aaggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3121 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3181 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3241 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     3301 cttaattaag acttctaata taacttcgta tagcatacat tatacgaagt tatggttcca
     3361 ctgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     3421 actaaaccca ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc
     3481 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3541 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3601 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3661 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3721 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3781 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3841 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3901 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3961 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4021 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4081 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4141 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4201 atcgccttct atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg
     4261 cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4321 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4381 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4441 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4501 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4561 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4621 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4681 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4741 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4801 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4861 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4921 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc
     4981 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5041 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg
     5101 gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg
     5161 ccatcgtgat ggctagagtt tagccctccc acacataacc agagggcagc aattcacgaa
     5221 tcccaactgc cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca
     5281 gatcgagaag cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc
     5341 cgatcgcgac gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca
     5401 cgagttctgc acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc
     5461 cgtcgctaga gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga
     5521 ttgtagccgt tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg
     5581 tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc
     5641 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
     5701 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     5761 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     5821 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     5881 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     5941 tccacagctg gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta
     6001 tttcaggcca tggtgctgcg c
//
