LOCUS       sequence-291170-        5000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of CteIF4E_35-250-del183-202-I87AI91A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5000)
  AUTHORS   Gruner S, Weber R, Peter D, Chung MY, Igreja C, Valkov E, Izaurralde
            E
  TITLE     Structural motifs in eIF4G and 4E-BPs modulate their binding to 
            eIF4E to regulate translation initiation in yeast.
  JOURNAL   Nucleic Acids Res. 2018 Jul 27;46(13):6893-6908. doi: 
            10.1093/nar/gky542.
  PUBMED    30053226
REFERENCE   2  (bases 1 to 5000)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2018-07-27- 27"; volume: "46"; issue: 
            "13"; pages: "6893-6908"
COMMENT     Sequence Label: pnYC-CvH-CteIF4E_35-250-del183-202-I87AI91A_AF
FEATURES             Location/Qualifiers
     source          1..5000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2293..2312
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2401..2420
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2401..2419
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2420..2444
                     /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             2459..2481
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3086..3106
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3134..3151
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3232..3250)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3246..3293
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3468..3473,3474..3490,3491..3496))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3608..4153
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4054..4073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4270..4287
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4679..4781
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4848..4867)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
       61 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
      121 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
      181 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
      241 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
      301 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
      361 ccgtctgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
      421 agtggcaggg cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctggtg
      481 ctacgcctga ataagtgata ataagcggat gaatggcaga aattcgaaag caaattcgac
      541 ccggtcgtcg gttcagggca gggtcgttaa atagccgctt atgtctattg ctggtttacc
      601 ggtttattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg aggccagttt
      661 gctcaggctc tccccgtgga ggtaataatt gacgatatga tcatttattc tgcctcccag
      721 agcctgataa aaacggttag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
      781 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgcttgt ttcggcgtgg
      841 gtatggtggc aggccccgtg gccgggggac tgttgggcgc tgccggcacc tgtcctacga
      901 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
      961 ggaaggagct accggacagc ggtgcggact gttgtaactc agaataagaa atgaggccgc
     1021 tcatggcgtt gactctcagt catagtatcg tggtatcacc ggttggttcc actctctgtt
     1081 gcgggcaact tcagcagcac gtaggggact tccgcgtttc cagactttac gaaacacgga
     1141 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     1201 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     1261 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     1321 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
     1381 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
     1441 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
     1501 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa
     1561 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccagtgat
     1621 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc
     1681 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
     1741 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
     1801 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     1861 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     1921 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     1981 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     2041 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgacg
     2101 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     2161 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     2221 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     2281 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     2341 gatgccggcc acgatgcgtc cggcgtagag gatcagacta agatctcgat cccgcgaaat
     2401 taatacgact cactataggg gaattgtgag cggataacaa ttccccacta gtaataattt
     2461 tgtttaactt taagaaggag atatacatat gaccgttttt cacgacaaag aaaacttcaa
     2521 cgttaaacat ccgctgagct gtcgttggac cctgtggttt accaaaccgg caagcggtaa
     2581 aggtgataat tggaatgatc tgctgaaaaa agtgatcacc tttgaaagcg tggaagaatt
     2641 ttggggcgcc tacaataacg cggcaccggt tagcgaactg gcagttaaaa gcgattacca
     2701 cctgtttaaa gagggtgttc gtccggaatg ggaagatccg cagaataaac atggtggcaa
     2761 atgggcatat cagttcaaag ataaacgcag cgtgaacatt gatgaactgt ggctgcatac
     2821 catgctggca gcaattggtg aaaccctgga agatgaagag gatggcgaag ttatgggtgt
     2881 tgttgttaat gtgcgcaaag gcttttatcg tattggtgtt tggacccgta ccaccgaaaa
     2941 aagcaaagaa atcctgatga atattggccg tcgtctgaaa gaagtgctga aactgcctcc
     3001 gaatgaaatg gttgaattta gcggtcatac cgaagcagca caggcaggta gcacccgtgc
     3061 aaaagcacgt atggttgttg gatccgaaaa cctttacttc cagggcagcg gcgccatggg
     3121 cagcggcagc ggccaccacc accaccacca cggtacctaa tagtctagag ctagccctag
     3181 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     3241 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     3301 gaggaactat atccggatat cccgcaagag gcccggcagt accggcataa ccaagcctat
     3361 gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat
     3421 acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc
     3481 gatgataagc tgtcaaacat gagaattaca acttatatcg tatggggctg acttcaggtg
     3541 ctacatttga agagataaat tgcactgaaa tcaagaaata ttttatctga ttaataagat
     3601 gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg
     3661 ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac cgaggtaact
     3721 ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg
     3781 acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg tgcttttgca
     3841 tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcggactga
     3901 acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga actgagtgtc
     3961 aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa accgaaaggc
     4021 aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc tttatagtcc
     4081 tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt caggggggcg
     4141 gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag tatcttcctg
     4201 gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg cagtcgaacg
     4261 accgagcgta gcgagtcagt gagcgaggaa gcggaatata tcctgtatca catattctgc
     4321 tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag cacttcactg acaccctcat
     4381 cagtgccaac atagtaagcc agtatacact cccctagcgc tgatgtccgg cggtgctttt
     4441 gccgttacgc accaccccgt cagtagctga acaggaggga cagctgatag aaacagaagc
     4501 cactggagca cctcaaaaac accatcatac actaaatcag taagttggca gcatcacccg
     4561 acgcactttg cgccgaataa atacctgtga cggaagatca cttcgcagaa taaataaatc
     4621 ctggtgtccc tgttgatacc gggaagccct gggccaactt ttggcgaaaa tgagacgttg
     4681 atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
     4741 tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     4801 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     4861 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     4921 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     4981 tgaatgctca tccggaattc
//
