LOCUS       sequence-290642-        6277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmThor-YMCLLtoAAAAA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6277)
  AUTHORS   Igreja C, Peter D, Weiler C, Izaurralde E
  TITLE     4E-BPs require non-canonical 4E-binding motifs and a lateral surface
            of eIF4E to repress translation.
  JOURNAL   Nat Commun. 2014 Sep 2;5:4790. doi: 10.1038/ncomms5790.
  PUBMED    25179781
REFERENCE   2  (bases 1 to 6277)
  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"; journalName: "Nat 
            Commun."; date: "2014-09-2"; volume: "5:4790. doi"; pages: " 
            10.1038/ncomms5790"
COMMENT     Sequence Label: pnEA-NvM-DmThor-YMCLLtoAAAAA_Z
FEATURES             Location/Qualifiers
     source          1..6277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      341..929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     830..849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1083..1100
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1115..1257
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1183..1202)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1343..1365
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1359..1550)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2862..3944)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3906..3925
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3945..4022)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4228..4247
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4331..4350
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4331..4349
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4350..4374
                     /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             4389..4411
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4418..5518
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="METEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             5534..5554
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(6004..6022)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6018..6065
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6240..6245,6246..6262,6263..6268))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
       61 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
      121 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
      181 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
      241 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
      301 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
      361 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
      421 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
      481 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
      541 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
      601 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
      661 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
      721 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
      781 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
      841 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
      901 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
      961 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     1021 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     1081 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     1141 atctgtgcgg tatttcacac cgcatatatg gtgcactctc agtacaatct gctctgatgc
     1201 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     1261 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     1321 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     1381 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     1441 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     1501 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     1561 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     1621 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     1681 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     1741 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     1801 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     1861 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     1921 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     1981 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
     2041 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg
     2101 gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat
     2161 ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc
     2221 gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt
     2281 tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca gtgatcgaag
     2341 ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct
     2401 gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca
     2461 taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt
     2521 cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     2581 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     2641 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     2701 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     2761 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     2821 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     2881 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2941 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     3001 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     3061 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     3121 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     3181 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     3241 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     3301 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     3361 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     3421 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     3481 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     3541 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     3601 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     3661 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     3721 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     3781 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     3841 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     3901 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     3961 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     4021 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     4081 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     4141 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     4201 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     4261 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat
     4321 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttccccacta
     4381 gtaataattt tgtttaactt taagaaggag atataccatg gaaactgaag aaggtaaact
     4441 ggtaatctgg attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt
     4501 cgagaaagat accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt
     4561 cccacaggtt gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt
     4621 tggtggctac gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga
     4681 caagctgtat ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc
     4741 gatcgctgtt gaagcgttat cgctgattta taacaaagac ctgctgccga acccgccaaa
     4801 aacctgggaa gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgccct
     4861 gatgttcaac ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta
     4921 tgcgttcaag tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg
     4981 cgcgaaagcg ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga
     5041 caccgattac tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa
     5101 cggcccgtgg gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact
     5161 gccgaccttc aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa
     5221 cgccgccagt ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga
     5281 tgaaggtctg gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta
     5341 cgaggaagag ttggcgaaag atccacgtat tgccgccacg atggaaaacg cccagaaagg
     5401 tgaaatcatg ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt
     5461 gatcaacgcc gccagcggtc gtcagactgt cgatgaagcc cttaaagacg cgcagactgg
     5521 taccggcagc ggcgaaaacc tttacttcca gggccatatg ctcgagatgt ccgcttcacc
     5581 caccgcccgt caagccatca cccaggccct gcccatgatc accaggaagg ttgtcatctc
     5641 ggatccgatc cagatgcccg aggtgtactc ctcgacgccc ggcggaaccc tctactccac
     5701 cactcctgga ggcaccaaac ttatcgccga gcgggctttc gcgaagaatc tccgtggctc
     5761 cccattgagc caaactccgc cgtccaacgt gcccagtgct gcggcgaggg gcactccgcg
     5821 tactcccttc cgcaagtgcg tgcccgtccc cacggaactg atcaagcaga ccaagtcgct
     5881 gaagatcgag gaccaggaac agttccaact ggatctgcaa ttgggatcct aatagtctag
     5941 agctagccct aggagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     6001 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     6061 ttttgctgaa aggaggaact atatccggat atcccgcaag aggcccggca gtaccggcat
     6121 aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt
     6181 gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca
     6241 ttaaagctta tcgatgataa gctgtcaaac atgagaa
//
