LOCUS       sequence-423368-        6065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CDC40. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6065)
  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 6065)
  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-C_CDC40:1-579
FEATURES             Location/Qualifiers
     source          1..6065
                     /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    3128..3249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3139..3158)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3364..3397)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3407..3454
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3486..4280
                     /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(3540..3559)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4150..4169
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4438..5026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4518..4537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5152..5200
                     /note="synthetic polyadenylation signal"
     CDS             complement(5224..5622)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5619..5628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5653..6010)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5667..5802
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5819..6010
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5967..5987
                     /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 ataggggcga tcgcaacaag cttcgggaag
     1201 atctggagct agcggcgggg gatccggtgg gaattcgaca ccatgtcggc tgcgattgca
     1261 gctctggccg cttcctatgg ttcgggttca gggtccgaat cggactcgga cagtgagagc
     1321 agtcggtgtc cgctgccagc cgccgactcc ctcatgcact tgactaaatc gccttcatca
     1381 aagccgtctc tagcagtggc agtggactcg gctccggagg tggcagttaa ggaagatttg
     1441 gagactggag ttcaccttga ccctgccgtc aaagaagttc agtataatcc tacctatgag
     1501 accatgtttg ctcctgagtt tggaccagaa aatcccttta ggacacagca aatggctgcc
     1561 cctagaaata tgctttctgg atatgccgaa ccagctcata tcaatgattt catgtttgag
     1621 cagcaaagga gaacttttgc aacatatggt tatgcattag acccttcatt agataatcat
     1681 caagtgtctg ctaaatatat tggttctgta gaagaagctg aaaaaaatca aggtttaact
     1741 gtatttgaaa ctggtcagaa gaaaacagaa aagaggaaaa agtttaaaga aaatgatgca
     1801 tccaatattg atggtttttt gggaccatgg gcaaaatatg tggatgaaaa agatgtagcc
     1861 aaaccttcag aagaagagca aaaagaattg gatgaaatca cagcaaagag gcagaaaaaa
     1921 ggaaaacagg aagaagagaa acctggggag gagaagacaa tcttacatgt taaagaaatg
     1981 tatgactatc aaggcaggtc ctatcttcac atacctcagg atgttggtgt taatctacgg
     2041 tcaactatgc cacctgagaa gtgttatctt cccaaaaaac aaattcatgt gtggtctgga
     2101 cacacaaagg gcgtcagtgc agtcagattg tttcctctct ctggccattt attgctgtct
     2161 tgttccatgg actgtaaaat taagctatgg gaggtttatg gagaacggcg ctgtctgaga
     2221 acatttattg gtcacagtaa ggctgttagg gatatctgct tcaatactgc aggaacacag
     2281 ttcctcagtg cagcctatga caggtatctt aagctctggg acactgagac aggacagtgt
     2341 atatcaagat ttacaaaccg aaaagtacct tattgtgtca aattcaatcc tgatgaagat
     2401 aagcaaaatc tctttgtggc tgggatgtct gataagaaga ttgtgcaatg ggacattcga
     2461 agtggagaaa ttgtgcagga atatgatcgg catttgggag ctgtcaacac cattgttttt
     2521 gtggatgaga ataggagatt tgtgagcaca tctgatgata aaagcctaag agtttgggaa
     2581 tgggatatcc ctgtggattt caagtacata gcagaaccca gtatgcactc aatgcctgca
     2641 gtgactttgt ctccaaatgg aaaatggcta gcatgccaat caatggacaa ccaaatctta
     2701 atttttggag cacagaacag atttagatta aataagaaaa aaatttttaa gggccatatg
     2761 gtagcaggct atgcttgtca ggtggacttt tcaccagaca tgagttatgt gatttcagga
     2821 gatggaaatg gaaaattaaa catttgggac tggaagacca caaaactcta cagtcgattt
     2881 aaagctcatg ataaagtgtg tataggtgca gtgtggcatc ctcatgaaac ttctaaggtc
     2941 ataacatgtg gttgggatgg tctcattaaa ttgtgggatg gctcgagcgg tgggagctcc
     3001 ggtgtgagcg gctggcggct gttcaagaag attagctaag tttaaacggc cgcgactcta
     3061 gactgcaggc atgcaagctg atttggctgc tgccaccgct gagcaaggcc gcttcgagca
     3121 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3181 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3241 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     3301 gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcttaat taagacttct
     3361 aatataactt cgtatagcat acattatacg aagttatggt tccactgttg acaattaatc
     3421 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa cccaggaggc
     3481 agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3541 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3601 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3661 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3721 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3781 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3841 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3901 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3961 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4021 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4081 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4141 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4201 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4261 ttcttgacga gttcttctga ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc
     4321 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4381 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4441 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4501 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4561 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4621 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4681 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4741 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4801 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4861 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4921 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4981 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat
     5041 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5101 gagattatca aaaaggatct tcacctagat ccttttatag tccggaatcg tcacacaaaa
     5161 aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg tgatggctag
     5221 agtttagccc tcccacacat aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg
     5281 ctgtccatca ctgtccttca ctatggcttt gatcccagga tgcagatcga gaagcacctg
     5341 tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca tttccgatcg cgacgataca
     5401 agtcaggttg ccagctgccg cagcagcagc agtgcccagc accacgagtt ctgcacaagg
     5461 tcccccagta aaatgatata cattgacacc agtgaagatg cggccgtcgc tagagagagc
     5521 tgcgctggcg acgctgtagt cttcagagat ggggatgctg ttgattgtag ccgttgctct
     5581 ttcaatgagg gtggattctt cttgagacaa aggcttggcc atggtggcgc tagtgtcaga
     5641 agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     5701 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     5761 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     5821 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     5881 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     5941 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
     6001 tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag gccatggtgc
     6061 tgcgc
//
