LOCUS       sequence-309816-        6264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression vector for the expression of N-terminal 
            6xHIS-SUMO-tagged codon optimized eIF4G1 amino acids 1-200 isoform 
            including the microexon..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6264)
  AUTHORS   Gonatopoulos-Pournatzis T, Niibori R, Salter EW, Weatheritt RJ, 
            Tsang B, Farhangmehr S, Liang X, Braunschweig U, Roth J, Zhang S, 
            Henderson T, Sharma E, Quesnel-Vallieres M, Permanyer J, Maier S, 
            Georgiou J, Irimia M, Sonenberg N, Forman-Kay JD, Gingras AC, 
            Collingridge GL, Woodin MA, Cordes SP, Blencowe BJ
  TITLE     Autism-Misregulated eIF4G Microexons Control Synaptic Translation 
            and Higher Order Cognitive Functions.
  JOURNAL   Mol Cell. 2020 Mar 19;77(6):1176-1192.e16. doi: 
            10.1016/j.molcel.2020.01.006. Epub 2020 Jan 29.
  PUBMED    31999954
REFERENCE   2  (bases 1 to 6264)
  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: 
            "10.1016/j.molcel.2020.01.006"; journalName: "Mol Cell"; date: 
            "2020-03-19- 19"; volume: "77"; issue: "6"; pages: "1176-1192.e16"
COMMENT     Sequence Label: PET.SUMO eIF4G1 +MIC 1-200
FEATURES             Location/Qualifiers
     source          1..6264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             101..118
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             128..145
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             152..445
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             1129..1146
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1199..1217)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1213..1260
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1297..1752
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1384..1403)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1594..1615
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1844..2659)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1844..2659)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2567..2586
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2651..2670
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2781..3369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3270..3289
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3523..3540
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3555..3694
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3620..3639)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3780..3802
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3796..3987)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4796..5878)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5840..5859
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5879..5956)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6162..6181
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gggcagcagc catcatcatc atcatcacgg
      121 cagcggcctg gtgccgcgcg gcagcgctag catgtcggac tcagaagtca atcaagaagc
      181 taagccagag gtcaagccag aagtcaagcc tgagactcac atcaatttaa aggtgtccga
      241 tggatcttca gagatcttct tcaagatcaa aaagaccact cctttaagaa ggctgatgga
      301 agcgttcgct aaaagacagg gtaaggaaat ggactcctta agattcttgt acgacggtat
      361 tagaattcaa gctgatcaga cccctgaaga tttggacatg gaggataacg atattattga
      421 ggctcacaga gaacagattg gtggtagtac tggatccaac aaggctccgc agagcactgg
      481 tccgcctccg gcaccttccc cggggcttcc tcagccggct tttccgcccg ggcagaccgc
      541 tcccgttgtt ttcagtactc cgcaagctac ccaaatgaac actccgtctc agccgcggca
      601 gggaggattt cggtctctgc agcatttcta cccgagcaga gcacagccgc ccagctcggc
      661 tgcaagcaga gttcagtcgg ctgcaccggc aagaccgggc ccggctgcac acgtataccc
      721 ggcaggcagc caagtaatga tgatcccgag ccagatcagc tacccggcaa gccagggagc
      781 atactacatc ccgggacagg gaaggagcac gtacgtagtc cccactcagc agtacccggt
      841 tcagccggga gcaccgggtt tttacccggg ggctagcccg actgaattcg gaacatacgc
      901 tggcgcatac tacccggcac aaggagttca gcagttcccg actggcgttg caccgacacc
      961 ggtactcatg aaccagccgc cgcagatagc tccgaaacgg gaaaggaaaa ctatcagaat
     1021 aagagacccg aaccaaggag gaaaagacat cacttaactc gagtgaagac aagcttaggt
     1081 atttattcgg cgcaaagtgc gtcgggtgat gctgccaact tagtcgagca ccaccaccac
     1141 caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     1201 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1261 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa
     1321 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     1381 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     1441 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     1501 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     1561 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     1621 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     1681 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     1741 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1801 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     1861 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     1921 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     1981 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     2041 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     2101 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     2161 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     2221 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     2281 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     2341 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     2401 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     2461 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     2521 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     2581 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     2641 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     2701 ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2761 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2821 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2881 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     2941 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3001 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3061 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3121 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3181 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3241 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3301 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3361 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3421 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3481 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3541 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3601 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     3661 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     3721 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3781 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     3841 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     3901 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     3961 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     4021 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     4081 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     4141 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     4201 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     4261 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     4321 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     4381 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     4441 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     4501 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     4561 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     4621 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     4681 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     4741 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     4801 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4861 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     4921 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     4981 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     5041 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     5101 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     5161 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     5221 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     5281 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     5341 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     5401 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     5461 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     5521 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     5581 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     5641 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     5701 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     5761 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     5821 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     5881 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     5941 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     6001 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     6061 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     6121 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     6181 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     6241 atcgagatct cgatcccgcg aaat
//
