LOCUS       sequence-425903-        5259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human EIF3I. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5259)
  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 5259)
  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_EIF3I:1-325
FEATURES             Location/Qualifiers
     source          1..5259
                     /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    2322..2443
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2333..2352)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2558..2591)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2601..2648
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2680..3474
                     /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(2734..2753)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3344..3363
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3632..4220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3712..3731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4346..4394
                     /note="synthetic polyadenylation signal"
     CDS             complement(4418..4816)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4813..4822
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4847..5204)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4861..4996
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5013..5204
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5161..5181
                     /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 tgaagccgat
     1261 cctactgcag ggccatgagc ggtccattac gcagattaag tataaccgcg aaggagacct
     1321 cctctttact gtggccaagg accctatcgt caatgtatgg tactctgtga atggtgagag
     1381 gctgggcacc tacatgggcc ataccggagc tgtgtggtgt gtggacgctg actgggacac
     1441 caagcatgtc ctcactggct cagctgacaa cagctgtcgt ctctgggact gtgaaacagg
     1501 aaagcagctg gcccttctca agaccaattc ggctgtccgg acctgcggtt ttgactttgg
     1561 gggcaacatc atcatgttct ccacggacaa gcagatgggc taccagtgct ttgtgagctt
     1621 ttttgacctg cgggatccga gccagattga caacaatgag ccctacatga agatcccttg
     1681 caatgactct aaaatcacca gtgctgtttg gggacccctg ggggagtgca tcatcgctgg
     1741 ccatgagagt ggagagctca accagtatag tgccaagtct ggagaggtgt tggtgaatgt
     1801 taaggagcac tcccggcaga tcaacgacat ccagttatcc agggacatga ccatgtttgt
     1861 gaccgcgtcc aaggacaaca cagccaagct ttttgactcc acaactcttg aacatcagaa
     1921 gactttccgg acagaacgtc ctgtcaactc agctgccctc tcccccaact atgaccatgt
     1981 ggtcctgggc ggtggtcagg aagccatgga tgtaaccaca acctccacca ggattggcaa
     2041 gtttgaggcc aggttcttcc atttggcctt tgaagaagag tttggaagag tcaagggtca
     2101 ctttggacct atcaacagtg ttgccttcca tcctgatggc aagagctaca gcagcggcgg
     2161 cgaagatggt tacgtccgta tccattactt cgacccacag tacttcgaat ttgagtttga
     2221 ggcttaatct agaggtcaag cttcgggggt ttaaacctgc aggcatgcaa gctgatttgg
     2281 ctgctgccac cgctgagcaa ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2341 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2401 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2461 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     2521 ctacaaatgt ggtaaaatct taattaagac ttctaatata acttcgtata gcatacatta
     2581 tacgaagtta tggttccact gttgacaatt aatcatcggc atagtatatc ggcatagtat
     2641 aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg
     2701 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     2761 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     2821 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     2881 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     2941 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3001 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3061 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3121 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3181 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     3241 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3301 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3361 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3421 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa
     3481 atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat
     3541 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3601 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3661 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3721 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3781 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3841 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3901 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3961 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4021 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4081 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4141 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4201 gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4261 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4321 agatcctttt atagtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat
     4381 aaagatattt tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag
     4441 agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg
     4501 ctttgatccc aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga
     4561 tgcccctgtt ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag
     4621 cagcagtgcc cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga
     4681 caccagtgaa gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag
     4741 agatggggat gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag
     4801 acaaaggctt ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta
     4861 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     4921 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     4981 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     5041 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     5101 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     5161 accctaactg acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag
     5221 ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
