LOCUS       sequence-423427-        5831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human EIF3B fragment V186 to T685. 
            C-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5831)
  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 5831)
  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_EIF3B:186-685
FEATURES             Location/Qualifiers
     source          1..5831
                     /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    2894..3015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2905..2924)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3130..3163)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3173..3220
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3252..4046
                     /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(3306..3325)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3916..3935
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4204..4792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4284..4303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4918..4966
                     /note="synthetic polyadenylation signal"
     CDS             complement(4990..5388)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5385..5394
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5419..5776)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5433..5568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5585..5776
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5733..5753
                     /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 ccatggtgat tgtagtggac
     1261 aatgtccctc aggtgggacc cgaccgactt gagaaactca aaaatgtcat ccacaagatc
     1321 ttttccaagt ttgggaaaat cacaaatgat ttttatcctg aagaggatgg gaagacaaaa
     1381 gggtatattt tcctggagta cgcgtcccct gcccacgctg tggatgctgt gaagaacgcc
     1441 gacggctaca agcttgacaa gcagcacaca ttccgggtca acctctttac ggattttgac
     1501 aagtatatga cgatcagtga cgagtgggat attccagaga aacagccttt caaagacctg
     1561 gggaacttac gttactggct tgaagaggca gaatgcagag atcagtacag tgtgattttt
     1621 gagagtggag accgcacttc catattctgg aatgacgtaa aagaccctgt ctcaattgaa
     1681 gaaagagcga gatggacaga gacgtatgtg cgttggtctc ctaagggcac ctacctggct
     1741 acctttcatc aaagaggcat tgctctatgg gggggagaga aattcaagca aattcagaga
     1801 ttcagccacc aaggggttca gcttattgac ttctcacctt gtgaaaggta cctggtgacc
     1861 tttagccccc tgatggacac gcaggatgac cctcaggcca taatcatctg ggacatcctt
     1921 acggggcaca agaagagggg ttttcactgt gagagctcag cccattggcc tatttttaag
     1981 tggagccatg atggcaaatt ctttgccaga atgaccctgg atacgcttag catctatgaa
     2041 actccttcta tgggtctttt ggacaagaag agtttgaaga tctctgggat aaaagacttt
     2101 tcttggtctc ctggtggtaa cataatcgcc ttctgggtgc ctgaagacaa agatattcca
     2161 gccagggtaa ccctgatgca gctccctacc aggcaagaga tccgagtgag gaacctgttc
     2221 aatgtggtgg actgcaagct ccattggcag aagaacggag actacttgtg tgtgaaagta
     2281 gataggactc cgaaaggcac ccagggtgtt gtcacaaatt ttgaaatttt ccgaatgagg
     2341 gagaaacagg tacctgtgga tgtggtcgag atgaaagaaa ccatcatagc ctttgcctgg
     2401 gaaccaaatg gaagtaagtt tgctgtgctg cacggagagg ctccgcggat atctgtgtct
     2461 ttctaccacg tcaaaaacaa cgggaagatt gaactcatca agatgttcga caagcagcag
     2521 gcgaacacca tcttctggag cccccaagga cagttcgtgg tgttggcggg cctgaggagt
     2581 atgaacggtg ccttagcgtt tgtggacact tcggactgca cggtcatgaa catcgcagag
     2641 cactacatgg cttccgacgt cgaatgggat cctactgggc gctacgtcgt cacctctgtg
     2701 tcctggtgga gccataaggt ggacaacgcg tactggctgt ggactggctc gagcggtggg
     2761 agctccggtg tgagcggctg gcggctgttc aagaagatta gctaagttta aacggccgcg
     2821 actctagact gcaggcatgc aagctgattt ggctgctgcc accgctgagc aaggccgctt
     2881 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2941 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     3001 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     3061 atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat cttaattaag
     3121 acttctaata taacttcgta tagcatacat tatacgaagt tatggttcca ctgttgacaa
     3181 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccca
     3241 ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3301 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3361 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3421 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3481 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3541 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3601 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3661 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3721 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3781 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3841 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3901 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3961 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4021 atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg cccaaccggt
     4081 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4141 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4201 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4261 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4321 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4381 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4441 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4501 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4561 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4621 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     4681 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4741 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     4801 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4861 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaatcgtcac
     4921 acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg ccatcgtgat
     4981 ggctagagtt tagccctccc acacataacc agagggcagc aattcacgaa tcccaactgc
     5041 cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca gatcgagaag
     5101 cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc cgatcgcgac
     5161 gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca cgagttctgc
     5221 acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc cgtcgctaga
     5281 gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga ttgtagccgt
     5341 tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg tggcgctagt
     5401 gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     5461 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     5521 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     5581 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5641 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
     5701 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg
     5761 gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta tttcaggcca
     5821 tggtgctgcg c
//
