LOCUS       sequence-291178-        4940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of SceIF4E_35-213.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4940)
  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 4940)
  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-NpH-SceIF4E_35-213_AF
FEATURES             Location/Qualifiers
     source          1..4940
                     /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(3172..3190)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3186..3233
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3408..3413,3414..3430,3431..3436))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3548..4093
                     /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     3994..4013
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4210..4227
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4619..4721
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4722..4940,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4788..4807)
                     /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 ttaaacatcc
     2581 gctgaatacc aaatggaccc tgtggtatac aaaaccggca gttgataaaa gcgaaagctg
     2641 gtcagatctg ctgcgtccgg ttaccagttt tcagaccgtt gaagaatttt gggccattat
     2701 tcagaatatt ccggaaccgc atgaactgcc gctgaaaagc gattatcatg tgtttcgtaa
     2761 tgatgttcgt ccggaatggg aagatgaagc aaatgcaaaa ggtggcaaat ggtcatttca
     2821 gctgcgtggt aaaggtgcag atattgatga actgtggctg cgtaccctgc tggcagttat
     2881 tggtgaaacc attgatgaag atgatagcca gattaatggt gtggttctga gcattcgtaa
     2941 aggcggtaac aaatttgcac tgtggaccaa aagcgaagat aaagaaccgc tgctgcgtat
     3001 tggtggtaaa ttcaaacagg ttctgaaact gaccgatgat ggtcacctgg aattttttcc
     3061 gcatagcagc gccaatggtc gtcatccgca gccgagcatt accctgtaag ctagccctag
     3121 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     3181 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     3241 gaggaactat atccggatat cccgcaagag gcccggcagt accggcataa ccaagcctat
     3301 gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat
     3361 acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc
     3421 gatgataagc tgtcaaacat gagaattaca acttatatcg tatggggctg acttcaggtg
     3481 ctacatttga agagataaat tgcactgaaa tcaagaaata ttttatctga ttaataagat
     3541 gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg
     3601 ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac cgaggtaact
     3661 ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg
     3721 acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg tgcttttgca
     3781 tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcggactga
     3841 acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga actgagtgtc
     3901 aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa accgaaaggc
     3961 aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc tttatagtcc
     4021 tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt caggggggcg
     4081 gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag tatcttcctg
     4141 gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg cagtcgaacg
     4201 accgagcgta gcgagtcagt gagcgaggaa gcggaatata tcctgtatca catattctgc
     4261 tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag cacttcactg acaccctcat
     4321 cagtgccaac atagtaagcc agtatacact cccctagcgc tgatgtccgg cggtgctttt
     4381 gccgttacgc accaccccgt cagtagctga acaggaggga cagctgatag aaacagaagc
     4441 cactggagca cctcaaaaac accatcatac actaaatcag taagttggca gcatcacccg
     4501 acgcactttg cgccgaataa atacctgtga cggaagatca cttcgcagaa taaataaatc
     4561 ctggtgtccc tgttgatacc gggaagccct gggccaactt ttggcgaaaa tgagacgttg
     4621 atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
     4681 tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     4741 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     4801 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     4861 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     4921 tgaatgctca tccggaattc
//
