LOCUS       sequence-290730-        3320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HseIF4E_36-217.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3320)
  AUTHORS   Peter D, Igreja C, Weber R, Wohlbold L, Weiler C, Ebertsch L, 
            Weichenrieder O, Izaurralde E
  TITLE     Molecular architecture of 4E-BP translational inhibitors bound to 
            eIF4E.
  JOURNAL   Mol Cell. 2015 Mar 19;57(6):1074-87. doi: 
            10.1016/j.molcel.2015.01.017. Epub 2015 Feb 19.
  PUBMED    25702871
REFERENCE   2  (bases 1 to 3320)
  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.2015.01.017"; journalName: "Mol Cell"; date: 
            "2015-03-19- 19"; volume: "57"; issue: "6"; pages: "1074-87"
COMMENT     Sequence Label: pnYC-NpH-HseIF4E_36-217_AA
FEATURES             Location/Qualifiers
     source          1..3320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     644..663
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     752..771
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        752..770
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    771..795
                     /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             810..832
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             851..868
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             890..913
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1552..1570)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1566..1613
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1788..1793,1794..1810,1811..1816))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      1928..2473
                     /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     2374..2393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2590..2607
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2999..3101
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3168..3187)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 atccttgaag ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg
       61 cgggcatccc gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc
      121 gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg
      181 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
      241 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
      301 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
      361 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
      421 ggttgaaggc tctcaagggc atcggtcgac gctctccctt atgcgactcc tgcattagga
      481 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
      541 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
      601 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
      661 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
      721 ggatcagact aagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
      781 gcggataaca attccccact agtaataatt ttgtttaact ttaagaagga gatataccat
      841 gggcagcagc catcatcatc atcatcacag cagcggtacc ggcagcggcc tggaagttct
      901 gttccagggg ccccatatga aacatcccct acagaacaga tgggcactct ggttttttaa
      961 aaatgataaa agcaaaactt ggcaagcaaa cctgcggctg atctccaagt ttgatactgt
     1021 tgaagacttt tgggctctgt acaaccatat ccagttgtct agtaatttaa tgcctggctg
     1081 tgactactca ctttttaagg atggtattga gcctatgtgg gaagatgaga aaaacaaacg
     1141 gggaggacga tggctaatta cattgaacaa acagcagaga cgaagtgacc tcgatcgctt
     1201 ttggctagag acacttctgt gccttattgg agaatctttt gatgactaca gtgatgatgt
     1261 atgtggcgct gttgttaatg ttagagctaa aggtgataag atagcaatat ggactactga
     1321 atgtgaaaac agagaagctg ttacacatat agggagggta tacaaggaaa ggttaggact
     1381 tcctccaaag atagtgattg gttatcagtc ccacgcagac acagctacta agagcggctc
     1441 caccactaaa aataggtttg ttgtttaacc gggatcctaa tagtctagag ctagccctag
     1501 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     1561 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     1621 gaggaactat atccggatat cccgcaagag gcccggcagt accggcataa ccaagcctat
     1681 gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat
     1741 acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc
     1801 gatgataagc tgtcaaacat gagaattaca acttatatcg tatggggctg acttcaggtg
     1861 ctacatttga agagataaat tgcactgaaa tcaagaaata ttttatctga ttaataagat
     1921 gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg
     1981 ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac cgaggtaact
     2041 ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg
     2101 acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg tgcttttgca
     2161 tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcggactga
     2221 acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga actgagtgtc
     2281 aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa accgaaaggc
     2341 aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc tttatagtcc
     2401 tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt caggggggcg
     2461 gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag tatcttcctg
     2521 gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg cagtcgaacg
     2581 accgagcgta gcgagtcagt gagcgaggaa gcggaatata tcctgtatca catattctgc
     2641 tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag cacttcactg acaccctcat
     2701 cagtgccaac atagtaagcc agtatacact cccctagcgc tgatgtccgg cggtgctttt
     2761 gccgttacgc accaccccgt cagtagctga acaggaggga cagctgatag aaacagaagc
     2821 cactggagca cctcaaaaac accatcatac actaaatcag taagttggca gcatcacccg
     2881 acgcactttg cgccgaataa atacctgtga cggaagatca cttcgcagaa taaataaatc
     2941 ctggtgtccc tgttgatacc gggaagccct gggccaactt ttggcgaaaa tgagacgttg
     3001 atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
     3061 tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     3121 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     3181 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     3241 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     3301 tgaatgctca tccggaattc
//
