LOCUS       sequence-496166-        7866 bp    DNA     circular SYN 05-JAN-2026
DEFINITION  Express the three subunits of human eIF2 in insect cells to purify 
            the complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7866)
  AUTHORS   Diaz-Lopez I, Gordiyenko Y, Zuber PK, Li X, Ramakrishnan V
  TITLE     Purification and characterization of recombinant human translation 
            initiation factor eIF3.
  JOURNAL   Protein Sci. 2026 Jan;35(1):e70388. doi: 10.1002/pro.70388.
  PUBMED    41432235
REFERENCE   2  (bases 1 to 7866)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 5, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Sci."; date: "2026-01"; volume: "35(1):e70388. doi"; pages: " 
            10.1002/pro.70388"
COMMENT     Sequence Label: pBIG1A_eIF2abg
FEATURES             Location/Qualifiers
     source          1..7866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1288..1379)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     polyA_signal    1535..1669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(2881..2972)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     polyA_signal    3128..3262
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(4420..4511)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     polyA_signal    4606..4654
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4679..4698
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     protein_bind    4744..4777
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(4848..4853,4854..4870,4871..4876)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5065..5598
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  5665..5888
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     rep_origin      complement(6023..6611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6103..6122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  6801..6966
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    7280..7414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7574..7597)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(7634..7657)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(7658..7681)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 acagggatcc tatgttcacc atgagcactt cattcttaga cagcttttga acctttgctg
       61 ctttcttgtc tccttcagtg cgtacaccta gaagccgtct aagcaggaaa taggaaattt
      121 ccaattctgt gaatatctca ggtaaagctc cgactgcacc aagtacttgc cccaccattc
      181 tgtcagcccg gcacaaagtg gggtcaattt ttgttccaac tccaataaga ccgcctggag
      241 cagcatattg cagatcatta tgctccgcaa aaagtgatac aattttggaa aagattggtt
      301 tacacatgag ttttccttca ctatctttgg aaacaatacc aggtcttact tctatctcct
      361 ggcccacctt taatactcct tttaggatac taccaccagc tacacctccc ttaaggtcat
      421 caacttcaca gccaggtttg ttgacatcaa aagatctaat aacaataagc cggggctctg
      481 aagtaaagtc tcttgggggt actggaattt tctttactat gtactcacaa acaacttcaa
      541 tattgtattt cagctgagct gaaattggaa taatgggagc tccctctgct actgtacctt
      601 ggacaaatgc aaggatctgc tcgtattgtt ctttagcctg actttctttt accaaatcaa
      661 ttttattttg tagaatcaaa atatgcttca gtttcatgat ctctatagca gccaggtgtt
      721 ccgatgtctg aggctgaggg caagattcat taccagctat caacagaaga gctgcatcca
      781 tcactgctgc accgttcagc atagtagcca tcaaaatatc gtggccagga cagtcaacaa
      841 aggaaacatg tctgactaat ttgaagttcc ctttggtccc tggaatgtcc gtaggaaact
      901 cgtcaggtgt actgctccca caagatctat aacattctgg ccgagggcaa cttgggtcat
      961 caagcttata aatcttagca ttagcatatc caagcttgat tgtaatattt ctttctagtt
     1021 catttttgaa cctgacagta tgaactccag aaatagcttt gacgactgtg gatttcccat
     1081 gagctacatg accaattgta cctatgttaa ttgtggcttg tctgctgata acttcgtgtg
     1141 aaagtggcgt caacttggta acatccaagg tggtgagatc ctgacgcgaa agatgcggct
     1201 gccctagagt cactccagct tctccgcccg ccatgttgac tcgagggatc cgcgcccgat
     1261 ggtgggacgg tatgaataat ccggaatatt tataggtttt tttattacaa aactgttacg
     1321 aaaacagtaa aatacttatt tatttgcgag atggttatca ttttaattat ctccatgatc
     1381 tattaatatt ccggagtagg tcgatttaaa catcaggcat cattaggttt atttaaatgg
     1441 ttatgatagt tattgctcag cggtggcagc agccaactca gcttcctttc gggctttgtt
     1501 agcagccgga tcttctaggc tcaagcagtg atcagatcca gacatgataa gatacattga
     1561 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1621 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1681 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     1741 aaacctctac aaatgtggta tggctgatta tgatcctcta gtacttctcg acaagcttgt
     1801 cgagactgca ggctctagag gtaccttagt tagctttggc acggagctgt gctcgcttgc
     1861 ccgtgacagc ctggaagccg gttttgatac tggcaacaga acatctagaa tgacaagttt
     1921 cgcactgtag gaaatagagt cgtgtgtcct tctgcaggat tgtgtccggt gatcggcatg
     1981 tgtgacaagt gacatattcc ttgatatatc ttctcaagac attttctatc tgtttctgtt
     2041 ggaatcttcc tttgattaca agttggttat taccatctat agaaccactt gtacccaatt
     2101 cagccaacaa aaatgcaagg agatgtttgg gctgacgatg taatagttta cagatatctg
     2161 taaagttgac aaaagaagtt ttcttggttc ctactcggac gacttgtgga ggtttcatga
     2221 caaatttcct tttctcccca gcaaccatat ctggattctt ttccctcatg atgttgaaca
     2281 ctcgattcag cagctcctcg tatgtgtagt ctctttctga gcctgcccaa gcagggcctg
     2341 tctgattact gaatgagata ccatcatctt ttttgttgtc ttcatcttct agagcttcat
     2401 ctttctctag tatttcatcc tcatctggga acttaacatt cttctttttc ttctttttat
     2461 tgccaagcat aatgtcaagg tcatcctctg gttcagttgg ttcttgaaca tcactttcaa
     2521 tcttaagatc ctttacacct tcttcagctt catcaatatc aaatatcttt ttagtttttt
     2581 tcttcttttt cttttgatta aagaagttca agtcatctag atcatcagaa gcatcttttt
     2641 tcctagtgtc ctcttcatca gcttccaaat ccttgtcctc agttggctct ggctccactt
     2701 cttttgtttc tgaaggctgg gtttcctctg tttgggtatc cccttcctca tctaacataa
     2761 aaggcttctt cttcttcttt ttcttcttgc tcatagtagg atcaaaaatc atctcgtccc
     2821 cagacatgtt gactcgaggg atccgcgccc gatggtggga cggtatgaat aatccggaat
     2881 atttataggt ttttttatta caaaactgtt acgaaaacag taaaatactt atttatttgc
     2941 gagatggtta tcattttaat tatctccatg atctattaat attccggagt aggtcgattt
     3001 aaacgtgcaa tagtatccag tttatttaaa tggttatgat agttattgct cagcggtggc
     3061 agcagccaac tcagcttcct ttcgggcttt gttagcagcc ggatcttcta ggctcaagca
     3121 gtgatcagat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3181 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3241 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3301 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     3361 ttatgatcct ctagtacttc tcgacaagct tgtcgagact gcaggctcta gaggtacctt
     3421 aatcttcagc tttggcttcc atttcttctg catcatcatc tccatccact tcggcatttt
     3481 ctctttcaag cctctccatc tgcctcgcaa gttcagtctc atctgtatct gtgaccactt
     3541 tgggctccat ttgaacattg aacacacccc tcttttcctc aatcttctct ttgataacag
     3601 ccatagcttg actgaggaca gaaaggcctt ctgttctctc cagggttgtc gtagtcatta
     3661 cataccgagg aggagctatt agattaatct taatgggcat gttttctgta gaacaattca
     3721 aacctgctct tagggcttct tttacagcat caatgccttc ataaccataa caagccactt
     3781 caatatctgc tcgaattttg acagcctgtg gggtcaagcg cctattaata ttattaatga
     3841 gtacttcccg ttcatcttca ttcaaatcta aactatccaa aatagatggg tctgagactg
     3901 catgcttaaa tgcatcatag gcaccatatc caggtctctt gtacttgtca tcaaagaccc
     3961 aggcagtcct ctggaatagg ctttccagct gctcatcctt ggtgtattct aacacctcag
     4021 caacatgacg aagaatgcta taaacagttt tggattttgt gaatttgtct tcacatttga
     4081 ttgcttcctc tggagaaact cttctttttg acaaatcaat atatcctttt tctttgtcca
     4141 ccctaatgac aaccacacac tcattcctgc caattcggat gagtttgttg atagaacgga
     4201 tacgccttct ggataattca ctaagaagaa tcatgccttc aatgttgttg tattccagca
     4261 agctgacata agcccccatt tcagcaatgg atctgacatt caccatcact acatcttcca
     4321 cctcaggaaa tttgtgttga taaaatctac aacttagacc cggcatgttg actcgaggga
     4381 tccgcgcccg atggtgggac ggtatgaata atccggaata tttataggtt tttttattac
     4441 aaaactgtta cgaaaacagt aaaatactta tttatttgcg agatggttat cattttaatt
     4501 atctccatga tctattaata ttccggagta ggtcgattta aatctttaga ccatagagcg
     4561 ttcacttgac agagtgccag ccctgggacc gaaccccgcg tttatgaaca aacgacccaa
     4621 cacccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc ttccggtatt
     4681 gtctccttcc gtgtttcagt tagcctcgat aaaaacttta gaccatagag cgtttaaacc
     4741 cagataactt cgtataatgt atgctatacg aagttatggc gacttcgctg ctgcccaagg
     4801 ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagttg acataagcct
     4861 gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
     4921 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     4981 tttttgtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
     5041 gtttgatgtt atggagcagc aacgatgtta cgcagcagca acgatgttac gcagcagggc
     5101 agtcgcccta aaacaaagtt aggtggctca agtatgggca tcattcgcac atgtaggctc
     5161 ggccctgacc aagtcaaatc catgcgggct gctcttgatc ttttcggtcg tgagttcgga
     5221 gacgtagcca cctactccca acatcagccg gactccgatt acctcgggaa cttgctccgt
     5281 agtaagacat tcatcgcgct tgctgccttc gaccaagaag cggttgttgg cgctctcgcg
     5341 gcttacgttc tgcccaagtt tgagcagccg cgtagtgaga tctatatcta tgatctcgca
     5401 gtctccggcg agcaccggag gcagggcatt gccaccgcgc tcatcaatct cctcaagcat
     5461 gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg tgacgatccc
     5521 gcagtggctc tctatacaaa gttgggcata cgggaagaag tgatgcactt tgatatcgac
     5581 ccaagtaccg ccacctaaca attcgttcaa gccgagatcg gcttcccggc cgcggagttg
     5641 ttcggtaaat tgtcacaacg ccgcgaatat agtctttaca tgcccttggc cacgcccctc
     5701 tttaatacga cgggcaattt gcacttcaga aaatgaagag tttgctttag ccataacaaa
     5761 agtccagtat gctttttcac agcataactg gactgatttc agtttacaac tattctgtct
     5821 agtttaagac tttattgtca tagtttagat ctattttgtt cagtttaaga ctttattgtc
     5881 cgcccacacc cgcttacgca gggcatctgc gcatccacag gaagagcgac ccaagtcaat
     5941 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6001 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6061 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6121 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6181 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6241 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6301 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6361 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6421 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6481 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6541 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6601 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6661 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6721 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6781 gttgacccaa gtcaaccggt tgtgggcgga caaaatagtt gggaactggg aggggtggaa
     6841 atggagtttt taaggattat ttagggaaga gtgacaaaat agatgggaac tgggtgtagc
     6901 gtcgtaagct aatacgaaaa ttaaaaatga caaaatagtt tggaactaga tttcacttat
     6961 ctggttcgga tctcctgttt aaaccccgat tgagatatag tttttaatta acccgtagtg
     7021 gctatggcag ggcttgccgc cccgacgttg gctgcgagcc ctgggccttc acccgaactt
     7081 gggggttggg gtggggaaaa ggaagaaacg cgggcgtatt ggtcccaatg gggtctcggt
     7141 ggggtatcga ctaaccccga ttgagatata gatttattta aatggttatg atagttattg
     7201 ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag ccggatcttc
     7261 taggctcaag cagtgatcag atccagacat gataagatac attgatgagt ttggacaaac
     7321 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     7381 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     7441 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     7501 tggtatggct gattatgatc ctctagtact tctcgacaag cttgtcgaga ctgcaggctc
     7561 tagaggtacc ttacttctca aactgagggt ggctccaacc agctccagat ccagcagcag
     7621 atccagcgga gcccttctcg aactgaggat ggctccaggg accttgaaac aggacttcga
     7681 gtccgtcatc atctactgtt ggcttgattg tcactcctct tcttatctga ccccaaccaa
     7741 ttaaacgcca gtgtttttca actcttcggc taagggcaat tttttctcct acctctgtgc
     7801 acactggatt ggtcaaaaca attttaccca aatcggcctt gacagcacta actctccctc
     7861 ctgttg
//
