LOCUS       sequence-423426-        6083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human EIF2A. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6083)
  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 6083)
  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_EIF2A:1-585
FEATURES             Location/Qualifiers
     source          1..6083
                     /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    3146..3267
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3157..3176)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3382..3415)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3425..3472
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3504..4298
                     /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(3558..3577)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4168..4187
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4456..5044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4536..4555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5170..5218
                     /note="synthetic polyadenylation signal"
     CDS             complement(5242..5640)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5637..5646
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5671..6028)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5685..5820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5837..6028
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5985..6005
                     /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 ccatggcgcc gtccacgccg
     1261 ctcttgacag tccgaggatc agaaggactg tacatggtga atggaccacc acattttaca
     1321 gaaagcacag tgtttccaag ggaatctggg aagaattgca aagtctgtat ctttagtaag
     1381 gatgggacct tgtttgcctg gggcaatgga gaaaaagtaa atattatcag tgtcactaac
     1441 aagggactac tgcactcctt cgacctcctg aaggcagttt gccttgaatt ctcacccaaa
     1501 aatactgtcc tggcaacgtg gcagccttac actacttcta aagatggcac agctgggata
     1561 cccaacctac aactttatga tgtgaaaact gggacatgtt tgaaatcttt catccagaaa
     1621 aaaatgcaaa attggtgtcc atcctggtca gaagatgaaa ctctttgtgc ccgcaatgtt
     1681 aacaatgaag ttcacttctt tgaaaacaac aattttaaca caattgcaaa taaattgcat
     1741 ttgcaaaaaa ttaatgactt tgtattatca cctggacccc aaccatacaa ggtggctgtc
     1801 tatgttccag gaagtaaagg tgcaccttca tttgttagat tatatcagta ccccaacttt
     1861 gctggacctc atgcagcttt agctaataaa agtttcttta aggcagataa agttacaatg
     1921 ctgtggaata aaaaagctac tgctgtgttg gtaatagcta gcacagatgt tgacaagaca
     1981 ggagcttcct actatggaga acaaactcta cactacattg caacaaatgg agaaagtgct
     2041 gtagtgcaat taccaaaaaa tggccccatt tatgatgtag tttggaattc tagttctact
     2101 gagttttgtg ctgtatatgg ttttatgcct gccaaagcga caattttcaa cttgaaatgt
     2161 gatcctgtat ttgactttgg aactggtcct cgtaatgcag cctactatag ccctcatgga
     2221 catatattag tattagctgg atttggaaat ctgaggggac aaatggaagt gtgggatgtg
     2281 aaaaactaca aacttatttc taaaccggtg gcttctgatt ctacatattt tgcttggtgc
     2341 ccggatggtg agcatatttt aacagctaca tgtgctccca ggttacgggt taataatgga
     2401 tacaaaattt ggcattatac tggctctatc ttgcacaagt atgatgtgcc atcaaatgca
     2461 gaattatggc aggtttcttg gcagccattt ttggatggaa tatttccagc aaaaacaata
     2521 acttaccaag cagttccaag tgaagtaccc aatgaggaac ctaaagttgc aacagcttat
     2581 agacccccag ctttaagaaa taaaccaatc accaattcca aattgcatga agaggaacca
     2641 cctcagaata tgaaaccaca atcaggaaac gataagccat tatcaaaaac agctcttaaa
     2701 aatcaaagga agcatgaagc taagaaagct gcaaagcagg aagcaagaag tgacaagagt
     2761 ccagatttgg cacctactcc tgccccacag agcacaccac gaaacactgt ctctcagtca
     2821 atttctgggg accctgagat agacaaaaaa atcaagaacc taaagaagaa actgaaagca
     2881 atcgaacaac tgaaagaaca agcagcaact ggaaaacagc tagaaaaaaa tcagttggag
     2941 aaaattcaga aagaaacagc ccttctccag gagctggaag atttggaatt gggtattggc
     3001 tcgagcggtg ggagctccgg tgtgagcggc tggcggctgt tcaagaagat tagctaagtt
     3061 taaacggccg cgactctaga ctgcaggcat gcaagctgat ttggctgctg ccaccgctga
     3121 gcaaggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     3181 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3241 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3301 gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa
     3361 atcttaatta agacttctaa tataacttcg tatagcatac attatacgaa gttatggttc
     3421 cactgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     3481 gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca ggttctccgg
     3541 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3601 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3661 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3721 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3781 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3841 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3901 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3961 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4021 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4081 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4141 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4201 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4261 gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt cgaaatgacc gaccaagcga
     4321 cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4381 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4441 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4501 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4561 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4621 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4681 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4741 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4801 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4861 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4921 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4981 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt
     5041 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5101 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc
     5161 cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga aaataaagat attttattgc
     5221 ggccatcgtg atggctagag tttagccctc ccacacataa ccagagggca gcaattcacg
     5281 aatcccaact gccgtcggct gtccatcact gtccttcact atggctttga tcccaggatg
     5341 cagatcgaga agcacctgtc ggcaccgtcc gcaggggctc aagatgcccc tgttctcatt
     5401 tccgatcgcg acgatacaag tcaggttgcc agctgccgca gcagcagcag tgcccagcac
     5461 cacgagttct gcacaaggtc ccccagtaaa atgatataca ttgacaccag tgaagatgcg
     5521 gccgtcgcta gagagagctg cgctggcgac gctgtagtct tcagagatgg ggatgctgtt
     5581 gattgtagcc gttgctcttt caatgagggt ggattcttct tgagacaaag gcttggccat
     5641 ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc
     5701 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     5761 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     5821 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     5881 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     5941 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     6001 attccacagc tggttctttc cgcctcagaa ggtacctaac caagttcctc tttcagaggt
     6061 tatttcaggc catggtgctg cgc
//
