LOCUS       sequence-291173-        4991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of CteIF4E_35-250-del183-202.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4991)
  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 4991)
  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-CteIF4E_35-250-del183-202_AF
FEATURES             Location/Qualifiers
     source          1..4991
                     /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(3223..3241)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3237..3284
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3459..3464,3465..3481,3482..3487))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3599..4144
                     /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     4045..4064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4261..4278
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4670..4772
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4773..4991,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4839..4858)
                     /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 ccccatatga ccgtttttca
     2581 cgacaaagaa aacttcaacg ttaaacatcc gctgagctgt cgttggaccc tgtggtttac
     2641 caaaccggca agcggtaaag gtgataattg gaatgatctg ctgaaaaaag tgatcacctt
     2701 tgaaagcgtg gaagaatttt ggggcatcta caataacatt gcaccggtta gcgaactggc
     2761 agttaaaagc gattaccacc tgtttaaaga gggtgttcgt ccggaatggg aagatccgca
     2821 gaataaacat ggtggcaaat gggcatatca gttcaaagat aaacgcagcg tgaacattga
     2881 tgaactgtgg ctgcatacca tgctggcagc aattggtgaa accctggaag atgaagagga
     2941 tggcgaagtt atgggtgttg ttgttaatgt gcgcaaaggc ttttatcgta ttggtgtttg
     3001 gacccgtacc accgaaaaaa gcaaagaaat cctgatgaat attggccgtc gtctgaaaga
     3061 agtgctgaaa ctgcctccga atgaaatggt tgaatttagc ggtcataccg aagcagcaca
     3121 ggcaggtagc acccgtgcaa aagcacgtat ggttgtttaa gctagcccta ggagatccgg
     3181 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     3241 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     3301 tatccggata tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc
     3361 atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc
     3421 ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttat cgatgataag
     3481 ctgtcaaaca tgagaattac aacttatatc gtatggggct gacttcaggt gctacatttg
     3541 aagagataaa ttgcactgaa atcaagaaat attttatctg attaataaga tgatcttctt
     3601 gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg
     3661 gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag
     3721 gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga
     3781 ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc
     3841 gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt
     3901 tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga
     3961 atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg
     4021 agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4081 tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg
     4141 gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc
     4201 aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt
     4261 agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg ctgacgcacc
     4321 ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca tcagtgccaa
     4381 catagtaagc cagtatacac tcccctagcg ctgatgtccg gcggtgcttt tgccgttacg
     4441 caccaccccg tcagtagctg aacaggaggg acagctgata gaaacagaag ccactggagc
     4501 acctcaaaaa caccatcata cactaaatca gtaagttggc agcatcaccc gacgcacttt
     4561 gcgccgaata aatacctgtg acggaagatc acttcgcaga ataaataaat cctggtgtcc
     4621 ctgttgatac cgggaagccc tgggccaact tttggcgaaa atgagacgtt gatcggcacg
     4681 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
     4741 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     4801 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     4861 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     4921 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     4981 atccggaatt c
//
