LOCUS       sequence-423434-        6773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human KCTD3. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6773)
  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 6773)
  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_KCTD3:1-815
FEATURES             Location/Qualifiers
     source          1..6773
                     /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    3836..3957
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3847..3866)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4072..4105)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4115..4162
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4194..4988
                     /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(4248..4267)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4858..4877
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5146..5734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5226..5245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5860..5908
                     /note="synthetic polyadenylation signal"
     CDS             complement(5932..6330)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6327..6336
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6361..6718)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6375..6510
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6527..6718
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6675..6695
                     /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 ccatggcggg agggcactgc
     1261 ggcagcttcc ccgcggcggc ggccggcagc ggcgagatcg tccaactgaa cgtagggggg
     1321 accagattta gtacctcaag acaaactctt atgtggattc cagattcttt tttttccagt
     1381 ttgctgagtg ggagaatttc aacacttcga gatgaaactg gtgctatatt tattgataga
     1441 gatccagcag catttgcacc cattttaaat tttcttcgga caaaagaact agacttaagg
     1501 ggagtgagta ttaatgttct caggcatgaa gcagaatttt acgggatcac tccattagta
     1561 agaaggcttc tcttatgtga agaattggag cgttcctctt gtggcagtgt cctttttcat
     1621 ggttacttgc ccccaccagg tattcctagt cgtaaaataa acaacacagt cagatctgct
     1681 gattctagga atggtctaaa ttctacagaa ggtgaagccc ggggaaatgg tacacagcct
     1741 gttctctctg gaacgggaga agaaactgtt aggctaggat ttcctgtgga tccacgaaag
     1801 gtgctaatag tagctggcca tcacaactgg attgtagctg catatgccca ttttgctgtg
     1861 tgttacagaa tcaaagaatc ttcaggatgg cagcaagtgt ttacgagccc atatttggat
     1921 tggactatcg aacgagtagc tttaaatgca aaggtggttg gagggccaca tggagacaaa
     1981 gacaaaatgg ttgctgttgc ctcagagagt agcatcatct tgtggagtgt tcaggatggg
     2041 ggaagtggaa gtgaaattgg agtgttcagc ctgggtgttc ctgtagatgc tctcttcttt
     2101 attggtaacc agttggtggc cacgagtcat acagggaaag tgggagtgtg gaatgctgtc
     2161 actcagcact ggcaggttca agatgttgtt cctataacta gttatgacac tgctggatca
     2221 ttccttctgc ttggatgtaa caatggatca atatattaca tagatatgca gaagttcccc
     2281 ttgcgaatga aagataatga tcttcttgta actgaactgt atcatgatcc ttcaaatgat
     2341 gctattactg ctctgagtgt ttacctcaca cccaaaacaa gtgtcagtgg taactggatc
     2401 gagatcgcct atggtacgag ctctggagca gtacgagtga ttgtacaaca cccagagaca
     2461 gttgggtcag gtcctcagct ttttcagact ttcacagttc accgaagtcc cgtaacaaaa
     2521 atcatgctat cagagaagca tcttgtatca gtctgtgcag ataataatca tgtccggacg
     2581 tggacagtaa cacgattcag aggaatgatc tctactcagc caggttctac tcctttagcg
     2641 tcattcaaga tactatccct ggaggagaca gaaagtcatg gtagctattc ctctggaaat
     2701 gacataggac cttttggaga gcgagacgat caacaggtgt ttatccagaa agttgttccc
     2761 atcaccaaca aactatttgt aagactctca tcgactggaa aaagaatatg tgagatccag
     2821 gctgttgact gtactacaat atcctcattt acagtgaggg aatgtgaggg atccagtagg
     2881 atgggctcaa gaccaaggcg ctacttgttc acaggccata caaatggcag tattcaaatg
     2941 tgggatctga ccactgctat ggatatggtt aacaaaagtg aagataagga tgtaggtggt
     3001 ccaaccgaag aagagctact caaattactc gatcaatgtg atttgagcac atctcgctgt
     3061 gctactccta acatcagtcc agcaacttcc gtagttcagc atagccactt acgagaatca
     3121 aattctagcc ttcagcttca gcaccatgat accacccatg aagcagctac ttacggttcc
     3181 atgaggcctt acagagaaag tcctttatta gcaagggcaa gaaggactga gagctttcac
     3241 agttataggg acttccagac tattaatttg aacagaaatg tagaaagagc tgtccctgaa
     3301 aatggtaact tgggtccaat acaagctgaa gtgaaagggg caacagggga atgtaatata
     3361 tctgagagaa agtctcctgg agtagaaata aaaagtttga gagaattgga tagtggattg
     3421 gaagtgcata aaatagctga aggtttttca gaatccaaga aaaggtcatc agaagatgaa
     3481 aatgaaaata aaatagagtt taggaagaaa ggaggatttg aagggggagg attccttgga
     3541 agaaagaaag ttccctatct ggcatcatca ccaagtactt ccgatggagg aactgactca
     3601 cctggtactg cgtccccatc tcctacaaag actactccat ctcctcggca taaaaaaagt
     3661 gattcttcag gtcaggagta cagcttgggc tcgagcggtg ggagctccgg tgtgagcggc
     3721 tggcggctgt tcaagaagat tagctaagtt taaacggccg cgactctaga ctgcaggcat
     3781 gcaagctgat ttggctgctg ccaccgctga gcaaggccgc ttcgagcaga catgataaga
     3841 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3901 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3961 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta
     4021 agcaagtaaa acctctacaa atgtggtaaa atcttaatta agacttctaa tataacttcg
     4081 tatagcatac attatacgaa gttatggttc cactgttgac aattaatcat cggcatagta
     4141 tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag atcatgattg
     4201 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4261 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4321 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4381 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4441 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4501 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4561 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4621 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4681 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4741 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4801 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4861 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4921 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4981 tcttctgatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc
     5041 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5101 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5161 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5221 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5281 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5341 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5401 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5461 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5521 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5581 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5641 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5701 gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg
     5761 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5821 aaggatcttc acctagatcc ttttatagtc cggaatcgtc acacaaaaaa ccaacacaca
     5881 gatgtaatga aaataaagat attttattgc ggccatcgtg atggctagag tttagccctc
     5941 ccacacataa ccagagggca gcaattcacg aatcccaact gccgtcggct gtccatcact
     6001 gtccttcact atggctttga tcccaggatg cagatcgaga agcacctgtc ggcaccgtcc
     6061 gcaggggctc aagatgcccc tgttctcatt tccgatcgcg acgatacaag tcaggttgcc
     6121 agctgccgca gcagcagcag tgcccagcac cacgagttct gcacaaggtc ccccagtaaa
     6181 atgatataca ttgacaccag tgaagatgcg gccgtcgcta gagagagctg cgctggcgac
     6241 gctgtagtct tcagagatgg ggatgctgtt gattgtagcc gttgctcttt caatgagggt
     6301 ggattcttct tgagacaaag gcttggccat ggtggcgcta gtgtcagaag aatcgagctt
     6361 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     6421 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     6481 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     6541 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     6601 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     6661 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagaa
     6721 ggtacctaac caagttcctc tttcagaggt tatttcaggc catggtgctg cgc
//
