LOCUS       sequence-290817-        4997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of CeeIF4E3_30-215opt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4997)
  AUTHORS   Peter D, Weber R, Kone C, Chung MY, Ebertsch L, Truffault V, 
            Weichenrieder O, Igreja C, Izaurralde E
  TITLE     Mextli proteins use both canonical bipartite and novel tripartite 
            binding modes to form eIF4E complexes that display differential 
            sensitivity to 4E-BP regulation.
  JOURNAL   Genes Dev. 2015 Sep 1;29(17):1835-49. doi: 10.1101/gad.269068.115. 
            Epub 2015 Aug 20.
  PUBMED    26294658
REFERENCE   2  (bases 1 to 4997)
  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: 
            "10.1101/gad.269068.115"; journalName: "Genes Dev"; date: 
            "2015-09-1- 1"; volume: "29"; issue: "17"; pages: "1835-49"
COMMENT     Sequence Label: pnYC-NpH-CeeIF4E3_30-215opt_AB
FEATURES             Location/Qualifiers
     source          1..4997
                     /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(3229..3247)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3243..3290
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3465..3470,3471..3487,3488..3493))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3605..4150
                     /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     4051..4070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4267..4284
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4676..4778
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4779..4997,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4845..4864)
                     /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 cccgtcatcc
     2581 gctgcagaat cgttgggcac tgtggtatct gaaagcagat cgtaataaag aatgggagga
     2641 ttgcctgaaa atggtgagcc tgtttgatac cgttgaagat ttttggagcc tgtataacca
     2701 tattcagagt gccggtggtc tgaattgggg tagcgattat tacctgttta aagaaggcat
     2761 taaaccgatg tgggaagatg tgaataatgt tcagggtggt cgttggctgg ttgttgttga
     2821 taaacagaaa ctgcagcgtc gtacccagct gctggatcat tattggctgg aactgctgat
     2881 ggccattgtt ggtgaacagt ttgatgaata tggcgattac atttgtggtg ccgttgttaa
     2941 tgttcgtcag aaaggtgata aagttagcct gtggacccgt gatgcaacac gtgatgatgt
     3001 gaatctgcgt attggtcagg ttctgaaaca aaaactgagc attccggata ccgaaattct
     3061 gcgttatgaa gtgcataaag atagcagcgc acgtaccagc agcaccgtta aaccgcgtat
     3121 ttgtctgtaa ctcgagctta agcaattggg atcctaatag tctagagcta gccctaggag
     3181 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     3241 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     3301 gaactatatc cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
     3361 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
     3421 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat
     3481 gataagctgt caaacatgag aattacaact tatatcgtat ggggctgact tcaggtgcta
     3541 catttgaaga gataaattgc actgaaatca agaaatattt tatctgatta ataagatgat
     3601 cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct
     3661 tgcagggcgg tttttcgaag gttctctgag ctaccaactc tttgaaccga ggtaactggc
     3721 ttggaggagc gcagtcacca aaacttgtcc tttcagttta gccttaaccg gcgcatgact
     3781 tcaagactaa ctcctctaaa tcaattacca gtggctgctg ccagtggtgc ttttgcatgt
     3841 ctttccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc ggactgaacg
     3901 gggggttcgt gcatacagtc cagcttggag cgaactgcct acccggaact gagtgtcagg
     3961 cgtggaatga gacaaacgcg gccataacag cggaatgaca ccggtaaacc gaaaggcagg
     4021 aacaggagag cgcacgaggg agccgccagg gggaaacgcc tggtatcttt atagtcctgt
     4081 cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgcttgtcag gggggcggag
     4141 cctatggaaa aacggctttg ccgcggccct ctcacttccc tgttaagtat cttcctggca
     4201 tcttccagga aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc
     4261 gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc tgtatcacat attctgctga
     4321 cgcaccggtg cagccttttt tctcctgcca catgaagcac ttcactgaca ccctcatcag
     4381 tgccaacata gtaagccagt atacactccc ctagcgctga tgtccggcgg tgcttttgcc
     4441 gttacgcacc accccgtcag tagctgaaca ggagggacag ctgatagaaa cagaagccac
     4501 tggagcacct caaaaacacc atcatacact aaatcagtaa gttggcagca tcacccgacg
     4561 cactttgcgc cgaataaata cctgtgacgg aagatcactt cgcagaataa ataaatcctg
     4621 gtgtccctgt tgataccggg aagccctggg ccaacttttg gcgaaaatga gacgttgatc
     4681 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     4741 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     4801 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     4861 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     4921 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     4981 atgctcatcc ggaattc
//
