LOCUS       sequence-425790-        6729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human KCTD3. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6729)
  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 6729)
  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_KCTD3:1-815
FEATURES             Location/Qualifiers
     source          1..6729
                     /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    3792..3913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3803..3822)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4028..4061)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4071..4118
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4150..4944
                     /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(4204..4223)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4814..4833
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5102..5690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5182..5201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5816..5864
                     /note="synthetic polyadenylation signal"
     CDS             complement(5888..6286)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6283..6292
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6317..6674)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6331..6466
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6483..6674
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6631..6651
                     /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 tggcgggagg
     1261 gcactgcggc agcttccccg cggcggcggc cggcagcggc gagatcgtcc aactgaacgt
     1321 aggggggacc agatttagta cctcaagaca aactcttatg tggattccag attctttttt
     1381 ttccagtttg ctgagtggga gaatttcaac acttcgagat gaaactggtg ctatatttat
     1441 tgatagagat ccagcagcat ttgcacccat tttaaatttt cttcggacaa aagaactaga
     1501 cttaagggga gtgagtatta atgttctcag gcatgaagca gaattttacg ggatcactcc
     1561 attagtaaga aggcttctct tatgtgaaga attggagcgt tcctcttgtg gcagtgtcct
     1621 ttttcatggt tacttgcccc caccaggtat tcctagtcgt aaaataaaca acacagtcag
     1681 atctgctgat tctaggaatg gtctaaattc tacagaaggt gaagcccggg gaaatggtac
     1741 acagcctgtt ctctctggaa cgggagaaga aactgttagg ctaggatttc ctgtggatcc
     1801 acgaaaggtg ctaatagtag ctggccatca caactggatt gtagctgcat atgcccattt
     1861 tgctgtgtgt tacagaatca aagaatcttc aggatggcag caagtgttta cgagcccata
     1921 tttggattgg actatcgaac gagtagcttt aaatgcaaag gtggttggag ggccacatgg
     1981 agacaaagac aaaatggttg ctgttgcctc agagagtagc atcatcttgt ggagtgttca
     2041 ggatggggga agtggaagtg aaattggagt gttcagcctg ggtgttcctg tagatgctct
     2101 cttctttatt ggtaaccagt tggtggccac gagtcataca gggaaagtgg gagtgtggaa
     2161 tgctgtcact cagcactggc aggttcaaga tgttgttcct ataactagtt atgacactgc
     2221 tggatcattc cttctgcttg gatgtaacaa tggatcaata tattacatag atatgcagaa
     2281 gttccccttg cgaatgaaag ataatgatct tcttgtaact gaactgtatc atgatccttc
     2341 aaatgatgct attactgctc tgagtgttta cctcacaccc aaaacaagtg tcagtggtaa
     2401 ctggatcgag atcgcctatg gtacgagctc tggagcagta cgagtgattg tacaacaccc
     2461 agagacagtt gggtcaggtc ctcagctttt tcagactttc acagttcacc gaagtcccgt
     2521 aacaaaaatc atgctatcag agaagcatct tgtatcagtc tgtgcagata ataatcatgt
     2581 ccggacgtgg acagtaacac gattcagagg aatgatctct actcagccag gttctactcc
     2641 tttagcgtca ttcaagatac tatccctgga ggagacagaa agtcatggta gctattcctc
     2701 tggaaatgac ataggacctt ttggagagcg agacgatcaa caggtgttta tccagaaagt
     2761 tgttcccatc accaacaaac tatttgtaag actctcatcg actggaaaaa gaatatgtga
     2821 gatccaggct gttgactgta ctacaatatc ctcatttaca gtgagggaat gtgagggatc
     2881 cagtaggatg ggctcaagac caaggcgcta cttgttcaca ggccatacaa atggcagtat
     2941 tcaaatgtgg gatctgacca ctgctatgga tatggttaac aaaagtgaag ataaggatgt
     3001 aggtggtcca accgaagaag agctactcaa attactcgat caatgtgatt tgagcacatc
     3061 tcgctgtgct actcctaaca tcagtccagc aacttccgta gttcagcata gccacttacg
     3121 agaatcaaat tctagccttc agcttcagca ccatgatacc acccatgaag cagctactta
     3181 cggttccatg aggccttaca gagaaagtcc tttattagca agggcaagaa ggactgagag
     3241 ctttcacagt tatagggact tccagactat taatttgaac agaaatgtag aaagagctgt
     3301 ccctgaaaat ggtaacttgg gtccaataca agctgaagtg aaaggggcaa caggggaatg
     3361 taatatatct gagagaaagt ctcctggagt agaaataaaa agtttgagag aattggatag
     3421 tggattggaa gtgcataaaa tagctgaagg tttttcagaa tccaagaaaa ggtcatcaga
     3481 agatgaaaat gaaaataaaa tagagtttag gaagaaagga ggatttgaag ggggaggatt
     3541 ccttggaaga aagaaagttc cctatctggc atcatcacca agtacttccg atggaggaac
     3601 tgactcacct ggtactgcgt ccccatctcc tacaaagact actccatctc ctcgacataa
     3661 aaaaagtgat tcttcaggtc aggagtacag cttgtaatct agaggtcaag cttcgggggt
     3721 ttaaacctgc aggcatgcaa gctgatttgg ctgctgccac cgctgagcaa ggccgcttcg
     3781 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3841 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3901 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     3961 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatct taattaagac
     4021 ttctaatata acttcgtata gcatacatta tacgaagtta tggttccact gttgacaatt
     4081 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg
     4141 aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4201 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4261 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4321 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4381 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4441 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4501 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4561 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4621 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4681 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4741 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4801 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4861 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4921 cgccttcttg acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat
     4981 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5041 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5101 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5161 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5221 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5281 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5341 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5401 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5461 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5521 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5581 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5641 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt
     5701 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5761 tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga atcgtcacac
     5821 aaaaaaccaa cacacagatg taatgaaaat aaagatattt tattgcggcc atcgtgatgg
     5881 ctagagttta gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg
     5941 tcggctgtcc atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca
     6001 cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga
     6061 tacaagtcag gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac
     6121 aaggtccccc agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga
     6181 gagctgcgct ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg
     6241 ctctttcaat gagggtggat tcttcttgag acaaaggctt ggccatggtg gcgctagtgt
     6301 cagaagaatc gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct
     6361 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc
     6421 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
     6481 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     6541 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc
     6601 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagctggt
     6661 tctttccgcc tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg
     6721 gtgctgcgc
//
