LOCUS       sequence-425901-        6039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human EIF2A. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6039)
  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 6039)
  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_EIF2A:1-585
FEATURES             Location/Qualifiers
     source          1..6039
                     /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    3102..3223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3113..3132)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3338..3371)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3381..3428
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3460..4254
                     /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(3514..3533)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4124..4143
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4412..5000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4492..4511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5126..5174
                     /note="synthetic polyadenylation signal"
     CDS             complement(5198..5596)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5593..5602
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5627..5984)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5641..5776
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5793..5984
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5941..5961
                     /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 tggcgccgtc
     1261 cacgccgctc ttgacagtcc gaggatcaga aggactgtac atggtgaatg gaccaccaca
     1321 ttttacagaa agcacagtgt ttccaaggga atctgggaag aattgcaaag tctgtatctt
     1381 tagtaaggat gggaccttgt ttgcctgggg caatggagaa aaagtaaata ttatcagtgt
     1441 cactaacaag ggactactgc actccttcga cctcctgaag gcagtttgcc ttgaattctc
     1501 acccaaaaat actgtcctgg caacgtggca gccttacact acttctaaag atggcacagc
     1561 tgggataccc aacctacaac tttatgatgt gaaaactggg acatgtttga aatctttcat
     1621 ccagaaaaaa atgcaaaatt ggtgtccatc ctggtcagaa gatgaaactc tttgtgcccg
     1681 caatgttaac aatgaagttc acttctttga aaacaacaat tttaacacaa ttgcaaataa
     1741 attgcatttg caaaaaatta atgactttgt attatcacct ggaccccaac catacaaggt
     1801 ggctgtctat gttccaggaa gtaaaggtgc accttcattt gttagattat atcagtaccc
     1861 caactttgct ggacctcatg cagctttagc taataaaagt ttctttaagg cagataaagt
     1921 tacaatgctg tggaataaaa aagctactgc tgtgttggta atagctagca cagatgttga
     1981 caagacagga gcttcctact atggagaaca aactctacac tacattgcaa caaatggaga
     2041 aagtgctgta gtgcaattac caaaaaatgg ccccatttat gatgtagttt ggaattctag
     2101 ttctactgag ttttgtgctg tatatggttt tatgcctgcc aaagcgacaa ttttcaactt
     2161 gaaatgtgat cctgtatttg actttggaac tggtcctcgt aatgcagcct actatagccc
     2221 tcatggacat atattagtat tagctggatt tggaaatctg aggggacaaa tggaagtgtg
     2281 ggatgtgaaa aactacaaac ttatttctaa accggtggct tctgattcta catattttgc
     2341 ttggtgcccg gatggtgagc atattttaac agctacatgt gctcccaggt tacgggttaa
     2401 taatggatac aaaatttggc attatactgg ctctatcttg cacaagtatg atgtgccatc
     2461 aaatgcagaa ttatggcagg tttcttggca gccatttttg gatggaatat ttccagcaaa
     2521 aacaataact taccaagcag ttccaagtga agtacccaat gaggaaccta aagttgcaac
     2581 agcttataga cccccagctt taagaaataa accaatcacc aattccaaat tgcatgaaga
     2641 ggaaccacct cagaatatga aaccacaatc aggaaacgat aagccattat caaaaacagc
     2701 tcttaaaaat caaaggaagc atgaagctaa gaaagctgca aagcaggaag caagaagtga
     2761 caagagtcca gatttggcac ctactcctgc cccacagagc acaccacgaa acactgtctc
     2821 tcagtcaatt tctggggacc ctgagataga caaaaaaatc aagaacctaa agaagaaact
     2881 gaaagcaatc gaacaactga aagaacaagc agcaactgga aaacagctag aaaaaaatca
     2941 gttggagaaa attcagaaag aaacagccct tctccaggag ctggaagatt tggaattggg
     3001 tatttaatct agaggtcaag cttcgggggt ttaaacctgc aggcatgcaa gctgatttgg
     3061 ctgctgccac cgctgagcaa ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3121 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3181 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3241 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     3301 ctacaaatgt ggtaaaatct taattaagac ttctaatata acttcgtata gcatacatta
     3361 tacgaagtta tggttccact gttgacaatt aatcatcggc atagtatatc ggcatagtat
     3421 aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg
     3481 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3541 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3601 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3661 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3721 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3781 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3841 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3901 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3961 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4021 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4081 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4141 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4201 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa
     4261 atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat
     4321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4981 gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5041 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5101 agatcctttt atagtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat
     5161 aaagatattt tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag
     5221 agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg
     5281 ctttgatccc aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga
     5341 tgcccctgtt ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag
     5401 cagcagtgcc cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga
     5461 caccagtgaa gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag
     5521 agatggggat gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag
     5581 acaaaggctt ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta
     5641 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     5701 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     5761 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     5821 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     5881 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     5941 accctaactg acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag
     6001 ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
