LOCUS       sequence-290965-        5057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmeIF4E_31-248.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  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 5057)
  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-NpH-DmeIF4E_31-248_AD
FEATURES             Location/Qualifiers
     source          1..5057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2294..2313
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2402..2421
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2402..2420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2421..2445
                     /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             2460..2482
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2501..2518
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2540..2563
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3289..3307)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3303..3350
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3525..3530,3531..3547,3548..3553))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3665..4210
                     /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     4111..4130
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4327..4344
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4736..4838
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4839..5057,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4905..4924)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ggtatggcaa 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 gccgtctgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      421 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
      481 gctacgcctg aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga
      541 cccggtcgtc ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac
      601 cggtttattg actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt
      661 tgctcaggct ctccccgtgg aggtaataat tgacgatatg atcatttatt ctgcctccca
      721 gagcctgata aaaacggtta gcgcttcgtt aatacagatg taggtgttcc acagggtagc
      781 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgcttg tttcggcgtg
      841 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ctgccggcac ctgtcctacg
      901 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
      961 cggaaggagc taccggacag cggtgcggac tgttgtaact cagaataaga aatgaggccg
     1021 ctcatggcgt tgactctcag tcatagtatc gtggtatcac cggttggttc cactctctgt
     1081 tgcgggcaac ttcagcagca cgtaggggac ttccgcgttt ccagacttta cgaaacacgg
     1141 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     1201 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     1261 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg
     1321 cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa
     1381 gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg
     1441 gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg
     1501 caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca
     1561 acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccagtga
     1621 tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat
     1681 ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa
     1741 gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca
     1801 gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     1861 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     1921 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     1981 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     2041 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac
     2101 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     2161 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     2221 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     2281 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     2341 tgatgccggc cacgatgcgt ccggcgtaga ggatcagact aagatctcga tcccgcgaaa
     2401 ttaatacgac tcactatagg ggaattgtga gcggataaca attccccact agtaataatt
     2461 ttgtttaact ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacag
     2521 cagcggtacc ggcagcggcc tggaagttct gttccagggg ccccatatgg ctaaggatgt
     2581 caagcccaag gaggacccac aggagactgg tgaaccagca ggcaacactg caaccactac
     2641 tgctcctgcc ggcgacgatg ctgtgcgcac cgagcattta tacaaacacc cgctcatgaa
     2701 tgtctggacg ctgtggtacc ttgaaaacga tcggtccaag tcctgggagg acatgcaaaa
     2761 cgagatcacc agcttcgata ccgtcgagga cttctggagc ctatacaacc acatcaagcc
     2821 cccatcagag atcaagctgg gtagtgacta ctcgctattc aagaagaaca ttcgtcccat
     2881 gtgggaggat gcagccaaca aacagggcgg tcgttgggtc attaccctta acaaaagctc
     2941 caagaccgat ctggataacc tatggctcga tgtgctgctc tgcctgattg gtgaggcctt
     3001 cgatcactct gatcagatct gcggcgctgt tataaacatt cgcggcaaga gcaacaagat
     3061 atccatctgg actgccgacg gaaacaacga ggaagctgcc cttgagattg gtcacaagct
     3121 gcgcgatgcc ctgcgtctgg gacgcaacaa ctcgctgcag tatcagttgc acaaggacac
     3181 gatggtcaag cagggctcca acgtgaaatc gatctacact ttgtaggcta gccctaggag
     3241 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     3301 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     3361 gaactatatc cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
     3421 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
     3481 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat
     3541 gataagctgt caaacatgag aattacaact tatatcgtat ggggctgact tcaggtgcta
     3601 catttgaaga gataaattgc actgaaatca agaaatattt tatctgatta ataagatgat
     3661 cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct
     3721 tgcagggcgg tttttcgaag gttctctgag ctaccaactc tttgaaccga ggtaactggc
     3781 ttggaggagc gcagtcacca aaacttgtcc tttcagttta gccttaaccg gcgcatgact
     3841 tcaagactaa ctcctctaaa tcaattacca gtggctgctg ccagtggtgc ttttgcatgt
     3901 ctttccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc ggactgaacg
     3961 gggggttcgt gcatacagtc cagcttggag cgaactgcct acccggaact gagtgtcagg
     4021 cgtggaatga gacaaacgcg gccataacag cggaatgaca ccggtaaacc gaaaggcagg
     4081 aacaggagag cgcacgaggg agccgccagg gggaaacgcc tggtatcttt atagtcctgt
     4141 cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgcttgtcag gggggcggag
     4201 cctatggaaa aacggctttg ccgcggccct ctcacttccc tgttaagtat cttcctggca
     4261 tcttccagga aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc
     4321 gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc tgtatcacat attctgctga
     4381 cgcaccggtg cagccttttt tctcctgcca catgaagcac ttcactgaca ccctcatcag
     4441 tgccaacata gtaagccagt atacactccc ctagcgctga tgtccggcgg tgcttttgcc
     4501 gttacgcacc accccgtcag tagctgaaca ggagggacag ctgatagaaa cagaagccac
     4561 tggagcacct caaaaacacc atcatacact aaatcagtaa gttggcagca tcacccgacg
     4621 cactttgcgc cgaataaata cctgtgacgg aagatcactt cgcagaataa ataaatcctg
     4681 gtgtccctgt tgataccggg aagccctggg ccaacttttg gcgaaaatga gacgttgatc
     4741 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     4801 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     4861 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     4921 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     4981 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     5041 atgctcatcc ggaattc
//
