LOCUS       sequence-422636-        5199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human SKIC8. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5199)
  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 5199)
  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_SKIC8:1-305
FEATURES             Location/Qualifiers
     source          1..5199
                     /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    2262..2383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2273..2292)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2498..2531)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2541..2588
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2620..3414
                     /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(2674..2693)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3284..3303
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3572..4160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3652..3671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4286..4334
                     /note="synthetic polyadenylation signal"
     CDS             complement(4358..4756)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4753..4762
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4787..5144)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4801..4936
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4953..5144
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5101..5121
                     /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 tgaccaacca
     1261 gtacggtatt ctcttcaaac aagagcaagc ccatgatgat gccatttggt cagttgcttg
     1321 ggggacaaac aagaaggaaa actctgagac agtggtcaca ggctccctag atgacctggt
     1381 gaaggtctgg aaatggcgtg atgagaggct ggacctacag tggagtctgg agggacatca
     1441 gctgggagtg gtgtctgtgg acatcagcca caccctgccc attgctgcat ccagctctct
     1501 tgatgctcat attcgtcttt gggacttgga aaatggcaaa cagataaagt ccatagatgc
     1561 aggacctgtg gatgcctgga ctttggcctt ttctcctgat tcccagtatc tggccacagg
     1621 aactcatgtc gggaaagtga acatttttgg tgtggaaagt gggaaaaagg aatattcttt
     1681 ggacacgaga ggaaaattca ttcttagtat tgcatatagt cctgatggga aatacctagc
     1741 cagtggagcc atagatggaa tcatcaatat ttttgatatt gcaactggaa aacttctgca
     1801 taccctggaa ggccatgcca tgcccattcg ctccttgacc ttttccccgg actcccagct
     1861 ccttgtcact gcttcagatg atggctacat caagatctat gatgtacaac atgccaattt
     1921 ggctggcacg ctgagcggcc atgcctcctg ggtgctgaac gttgcattct gtcctgatga
     1981 cactcacttt gtttccagtt cgtctgacaa aagtgtaaaa gtttgggatg ttggaacgag
     2041 gacttgtgtt cacaccttct ttgatcacca ggatcaggtc tggggagtaa aatacaatgg
     2101 aaatggttca aaaattgtgt ctgttggaga tgaccaggaa attcacatct atgattgtcc
     2161 aatttaatct agaggtcaag cttcgggggt ttaaacctgc aggcatgcaa gctgatttgg
     2221 ctgctgccac cgctgagcaa ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2281 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2341 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2401 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     2461 ctacaaatgt ggtaaaatct taattaagac ttctaatata acttcgtata gcatacatta
     2521 tacgaagtta tggttccact gttgacaatt aatcatcggc atagtatatc ggcatagtat
     2581 aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg
     2641 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     2701 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     2761 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     2821 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     2881 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     2941 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3001 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3061 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3121 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     3181 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3241 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3301 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3361 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa
     3421 atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat
     3481 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3541 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3601 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3661 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3721 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3781 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3841 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3901 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3961 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4021 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4081 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4141 gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4201 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4261 agatcctttt atagtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat
     4321 aaagatattt tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag
     4381 agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg
     4441 ctttgatccc aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga
     4501 tgcccctgtt ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag
     4561 cagcagtgcc cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga
     4621 caccagtgaa gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag
     4681 agatggggat gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag
     4741 acaaaggctt ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta
     4801 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     4861 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     4921 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     4981 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     5041 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     5101 accctaactg acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag
     5161 ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
