LOCUS       sequence-358057-        4560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HseIF4E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4560)
  AUTHORS   Gruner S, Peter D, Weber R, Wohlbold L, Chung MY, Weichenrieder O, 
            Valkov E, Igreja C, Izaurralde E
  TITLE     The Structures of eIF4E-eIF4G Complexes Reveal an Extended Interface
            to Regulate Translation Initiation.
  JOURNAL   Mol Cell. 2016 Nov 3;64(3):467-479. doi: 
            10.1016/j.molcel.2016.09.020. Epub 2016 Oct 20.
  PUBMED    27773676
REFERENCE   2  (bases 1 to 4560)
  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.2016.09.020"; journalName: "Mol Cell"; date: 
            "2016-11-3- 3"; volume: "64"; issue: "3"; pages: "467-479"
COMMENT     Sequence Label: pnYC-HseIF4E_AF
FEATURES             Location/Qualifiers
     source          1..4560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1853..1872
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1961..1980
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1961..1979
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1980..2004
                     /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             2019..2041
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2792..2810)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2806..2853
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3028..3033,3034..3050,3051..3056))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3168..3713
                     /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     3614..3633
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3830..3847
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4239..4341
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4342..4560,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4408..4427)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
       61 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
      121 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
      181 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
      241 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
      301 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
      361 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      421 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
      481 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg
      541 acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta
      601 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt
      661 ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcatttat tctgcctccc
      721 agagcctgat aaaaacggtt agcgcttcgt taatacagat gtaggtgttc cacagggtag
      781 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctt gtttcggcgt
      841 gggtatggtg gcaggccccg tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg
      901 ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca
      961 gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc
     1021 cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga
     1081 ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg
     1141 aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg ctggtaagag
     1201 ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg
     1261 cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca
     1321 tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg
     1381 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     1441 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     1501 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     1561 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     1621 agctgactgg gttgaaggct ctcaagggca tcggtcgacg ctctccctta tgcgactcct
     1681 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     1741 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     1801 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     1861 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     1921 cggcgtagag gatcagacta agatctcgat cccgcgaaat taatacgact cactataggg
     1981 gaattgtgag cggataacaa ttccccacta gtaataattt tgtttaactt taagaaggag
     2041 atatacatat gctcgagatg gcgaccgtcg aaccggaaac cacccctact cctaatcccc
     2101 cgactacaga agaggagaaa acggaatcta atcaggaggt tgctaaccca gaacactata
     2161 ttaaacatcc cctacagaac agatgggcac tctggttttt taaaaatgat aaaagcaaaa
     2221 cttggcaagc aaacctgcgg ctgatctcca agtttgatac tgttgaagac ttttgggctc
     2281 tgtacaacca tatccagttg tctagtaatt taatgcctgg ctgtgactac tcacttttta
     2341 aggatggtat tgagcctatg tgggaagatg agaaaaacaa acggggagga cgatggctaa
     2401 ttacattgaa caaacagcag agacgaagtg acctcgatcg cttttggcta gagacacttc
     2461 tgtgccttat tggagaatct tttgatgact acagtgatga tgtatgtggc gctgttgtta
     2521 atgttagagc taaaggtgat aagatagcaa tatggactac tgaatgtgaa aacagagaag
     2581 ctgttacaca tatagggagg gtatacaagg aaaggttagg acttcctcca aagatagtga
     2641 ttggttatca gtcccacgca gacacagcta ctaagagcgg ctccaccact aaaaataggt
     2701 ttgttgttta aggatcctaa tagtctagag ctagccctag gagatccggc tgctaacaaa
     2761 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     2821 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatat
     2881 cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga
     2941 cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc tgactgcgtt
     3001 agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc tgtcaaacat
     3061 gagaattaca acttatatcg tatggggctg acttcaggtg ctacatttga agagataaat
     3121 tgcactgaaa tcaagaaata ttttatctga ttaataagat gatcttcttg agatcgtttt
     3181 ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg
     3241 aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca
     3301 ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct
     3361 aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc
     3421 aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca
     3481 gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac
     3541 gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga
     3601 gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact
     3661 gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct
     3721 ttgccgcggc cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc
     3781 gccccgttcg taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt
     3841 gagcgaggaa gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt
     3901 ttttctcctg ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc
     3961 agtatacact cccctagcgc tgatgtccgg cggtgctttt gccgttacgc accaccccgt
     4021 cagtagctga acaggaggga cagctgatag aaacagaagc cactggagca cctcaaaaac
     4081 accatcatac actaaatcag taagttggca gcatcacccg acgcactttg cgccgaataa
     4141 atacctgtga cggaagatca cttcgcagaa taaataaatc ctggtgtccc tgttgatacc
     4201 gggaagccct gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc
     4261 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
     4321 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
     4381 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     4441 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
     4501 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc
//
