LOCUS       sequence-291171-        4985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of CteIF4E_40-250-del183-202.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4985)
  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 4985)
  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_40-250-del183-202_AF
FEATURES             Location/Qualifiers
     source          1..4985
                     /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             3071..3091
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3119..3136
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3217..3235)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3231..3278
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3453..3458,3459..3475,3476..3481))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3593..4138
                     /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     4039..4058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4255..4272
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4664..4766
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4833..4852)
                     /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 gaaagaaaac ttcaacgtta aacatccgct
     2521 gagctgtcgt tggaccctgt ggtttaccaa accggcaagc ggtaaaggtg ataattggaa
     2581 tgatctgctg aaaaaagtga tcacctttga aagcgtggaa gaattttggg gcatctacaa
     2641 taacattgca ccggttagcg aactggcagt taaaagcgat taccacctgt ttaaagaggg
     2701 tgttcgtccg gaatgggaag atccgcagaa taaacatggt ggcaaatggg catatcagtt
     2761 caaagataaa cgcagcgtga acattgatga actgtggctg cataccatgc tggcagcaat
     2821 tggtgaaacc ctggaagatg aagaggatgg cgaagttatg ggtgttgttg ttaatgtgcg
     2881 caaaggcttt tatcgtattg gtgtttggac ccgtaccacc gaaaaaagca aagaaatcct
     2941 gatgaatatt ggccgtcgtc tgaaagaagt gctgaaactg cctccgaatg aaatggttga
     3001 atttagcggt cataccgaag cagcacaggc aggtagcacc cgtgcaaaag cacgtatggt
     3061 tgttggatcc gaaaaccttt acttccaggg cagcggcgcc atgggcagcg gcagcggcca
     3121 ccaccaccac caccacggta cctaatagtc tagagctagc cctaggagat ccggctgcta
     3181 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     3241 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     3301 gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag
     3361 ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact
     3421 gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga taagctgtca
     3481 aacatgagaa ttacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga
     3541 taaattgcac tgaaatcaag aaatatttta tctgattaat aagatgatct tcttgagatc
     3601 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt
     3661 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
     3721 agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
     3781 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
     3841 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
     3901 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
     3961 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
     4021 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4081 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
     4141 cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa
     4201 tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag
     4261 tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg caccggtgca
     4321 gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg ccaacatagt
     4381 aagccagtat acactcccct agcgctgatg tccggcggtg cttttgccgt tacgcaccac
     4441 cccgtcagta gctgaacagg agggacagct gatagaaaca gaagccactg gagcacctca
     4501 aaaacaccat catacactaa atcagtaagt tggcagcatc acccgacgca ctttgcgccg
     4561 aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg
     4621 ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag
     4681 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga
     4741 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     4801 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     4861 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     4921 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     4981 aattc
//
