LOCUS       sequence-425885-        5345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CIAO1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5345)
  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 5345)
  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_CIAO1:1-339
FEATURES             Location/Qualifiers
     source          1..5345
                     /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    2408..2529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2419..2438)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2644..2677)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2687..2734
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2766..3560
                     /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(2820..2839)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3430..3449
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3718..4306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3798..3817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4432..4480
                     /note="synthetic polyadenylation signal"
     CDS             complement(4504..4902)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4899..4908
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4933..5290)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4947..5082
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5099..5290
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5247..5267
                     /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 ccatgaagga ctcgctggtg
     1261 ctgctgggcc gtgtcccggc gcacccggac tcccgctgct ggttcctggc ctggaacccc
     1321 gcggggaccc tgctggcctc gtgcggcggc gaccggagaa tccgcatctg gggcacggag
     1381 ggtgacagct ggatctgcaa gtctgtcctt tctgaaggcc accagcgcac cgtgcggaag
     1441 gtagcctggt ccccctgcgg taattacctg gcctctgcca gctttgatgc taccacttgc
     1501 atttggaaga agaaccagga tgactttgag tgtgtaacca ctctcgaggg ccatgaaaat
     1561 gaggtcaagt cagtggcttg ggccccatct ggcaacctcc tggccacttg cagccgagat
     1621 aagagcgttt gggtctggga agttgatgaa gaggatgagt atgaatgtgt cagtgttctc
     1681 aactcccaca cacaggatgt caagcatgtg gtttggcacc caagtcagga gctcttagct
     1741 tctgccagct atgatgacac agtgaagctg taccgggagg aagaggatga ctgggtatgc
     1801 tgtgccaccc ttgagggcca tgaatccact gtgtggagct tggcctttga cccgagtggc
     1861 cagcgcctgg cgtcttgtag tgatgaccgt actgtgcgta tctggcgtca gtatctacca
     1921 ggcaatgaac aaggggtggc atgcagcggc tctgacccca gttggaaatg tatctgtact
     1981 ttgtccggct tccactcaag gaccatttat gacattgctt ggtgtcagct gacaggggct
     2041 ctggccacag cttgtgggga tgacgcgatc cgcgtgtttc aggaggatcc caactcggat
     2101 ccacagcagc ccaccttctc cctgacagcc cacttgcatc aggcccattc ccaggatgtc
     2161 aactgtgtgg cctggaaccc caaggagcca gggctactgg cctcctgcag tgatgatggg
     2221 gaggtggcct tctggaagta tcagcggcct gaaggcctcg gctcgagcgg tgggagctcc
     2281 ggtgtgagcg gctggcggct gttcaagaag attagctaag tttaaacggc cgcgactcta
     2341 gactgcaggc atgcaagctg atttggctgc tgccaccgct gagcaaggcc gcttcgagca
     2401 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2461 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2521 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     2581 gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcttaat taagacttct
     2641 aatataactt cgtatagcat acattatacg aagttatggt tccactgttg acaattaatc
     2701 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa cccaggaggc
     2761 agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     2821 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     2881 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     2941 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3001 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3061 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3121 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3181 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3241 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     3301 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3361 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3421 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3481 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3541 ttcttgacga gttcttctga ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc
     3601 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3661 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3721 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3781 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3841 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3901 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3961 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4021 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4081 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4141 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4201 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4261 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat
     4321 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4381 gagattatca aaaaggatct tcacctagat ccttttatag tccggaatcg tcacacaaaa
     4441 aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg tgatggctag
     4501 agtttagccc tcccacacat aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg
     4561 ctgtccatca ctgtccttca ctatggcttt gatcccagga tgcagatcga gaagcacctg
     4621 tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca tttccgatcg cgacgataca
     4681 agtcaggttg ccagctgccg cagcagcagc agtgcccagc accacgagtt ctgcacaagg
     4741 tcccccagta aaatgatata cattgacacc agtgaagatg cggccgtcgc tagagagagc
     4801 tgcgctggcg acgctgtagt cttcagagat ggggatgctg ttgattgtag ccgttgctct
     4861 ttcaatgagg gtggattctt cttgagacaa aggcttggcc atggtggcgc tagtgtcaga
     4921 agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     4981 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     5041 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     5101 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     5161 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     5221 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
     5281 tccgcctcag aaggtaccta accaagttcc tctttcagag gttatttcag gccatggtgc
     5341 tgcgc
//
