LOCUS       sequence-288661-        6859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsEDC3_192-508-HsRck-K135A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6859)
  AUTHORS   Tritschler F, Braun JE, Eulalio A, Truffault V, Izaurralde E, 
            Weichenrieder O
  TITLE     Structural basis for the mutually exclusive anchoring of P body 
            components EDC3 and Tral to the DEAD box protein DDX6/Me31B.
  JOURNAL   Mol Cell. 2009 Mar 13. 33(5):661-8.
  PUBMED    19285948
REFERENCE   2  (bases 1 to 6859)
  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: "Mol Cell. 
            2009 Mar 13. 33(5):661-8."
COMMENT     Sequence Label: pRSFDuetP-HsEDC3_192-508-GST-HsRCK-K135A_E
FEATURES             Location/Qualifiers
     source          1..6859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             110..133
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        1168..1186
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1187..1211
                     /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)."
     CDS             1272..1925
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1883..1905
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1932..1955
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3396..3440
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3478..3496)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3492..3539
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3769..4584)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3769..4584)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4492..4511
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4585..4676)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4692..5441)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(4702..4719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4942..4961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5637..6719)
                     /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     6681..6700
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6720..6797)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6843..6859,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agcgcgggtc tggaagttct
      121 gttccagggg ccccaggatc cgaattcgtt cggggatgat attgaggaga tcccagacac
      181 agattttgat tttgaaggga acctggctct ttttgacaag gcagctgtgt ttgaggagat
      241 tgatacctat gaaaggagaa gtggtacccg ttcccggggc atcccaaatg aaaggcccac
      301 tcggtaccgc catgatgaga acatcttgga gtccgagccc attgtctatc gacggatcat
      361 agtgccccac aacgtgagca aggagttctg cacggactct ggcctggttg tcccaagtat
      421 ttcctatgag ctgcataaaa agctgttgtc cgtggctgag aagcatgggc tgacccttga
      481 gcggagactg gagatgacag gtgtgtgtgc cagtcagatg gcactgaccc tcctcggagg
      541 acctaacagg ttgaatccca aaaatgttca ccagaggcct acagtggctc tactgtgtgg
      601 acctcatgtg aagggggctc agggtatcag ctgtggaagg cacctagcca accatgatgt
      661 ccaggtcatc cttttcctgc ccaattttgt caagatgttg gaatctatca ccaatgagct
      721 gtcgctcttc agcaagaccc aaggccaaca agtgtctagc ctcaaagatc tgcccactag
      781 ccctgtggac ctggtcatca actgcctgga ttgccctgag aacgtcttcc tgcgcgatca
      841 accctggtac aaggcagctg tggcctgggc caaccagaac cgggcaccag tactcagcat
      901 agaccctcct gtgcatgaag tcgaacaggg cattgatgcc aaatggtcac tggcactggg
      961 cctgcctctg ccactggggg agcacgcagg ccgtatctat ttgtgcgaca ttggcattcc
     1021 ccagcaggtc ttccaggagg tgggcatcaa ctaccactcg ccctttggct gcaagtttgt
     1081 tatcccactg cactctgctt aggcggccgc ataatgctta agtcgaacag aaagtaatcg
     1141 tattgtacac ggccgcataa tcgaaattaa tacgactcac tataggggaa ttgtgagcgg
     1201 ataacaattc cccatcttag tatattagtt aagtataaga aggagatata catatggcag
     1261 atctcaattg gatgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca
     1321 ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg
     1381 aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt
     1441 attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg
     1501 acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag
     1561 gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa
     1621 ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt
     1681 tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg
     1741 acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag
     1801 tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca
     1861 agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc
     1921 caaaatcgga tctggaagtt ctgttccagg ggcccctggg atccatgggt ctgtccagtc
     1981 aaaatggtca gctgagaggc cctgtgaaac ccactggtgg ccctggagga gggggcacac
     2041 agacacagca acagatgaac cagctgaaaa acaccaacac aatcaataat ggcactcagc
     2101 agcaagcaca gagtatgacc accactatta aacctggtga tgactggaaa aagactttaa
     2161 aactccctcc aaaggatcta agaatcaaaa cttcggatgt gacctccaca aaaggaaatg
     2221 agtttgaaga ttactgtttg aaacgggagt tactgatggg aatttttgaa atgggctggg
     2281 aaaagccatc tcctattcag gaggagagca ttcccattgc tttatctggt agggatatct
     2341 tagctagagc aaaaaatgga acaggcgcaa gcggtgccta cctcattccc ttacttgaac
     2401 ggctagacct gaagaaggac aatatacaag caatggtgat tgttcccact agagaacttg
     2461 ctctacaggt cagtcaaatt tgcatccagg tcagcaaaca catgggaggg gccaaagtga
     2521 tggcaaccac aggaggaacc aatttacgag atgacataat gaggcttgat gatacagtgc
     2581 acgtggtgat tgctacccct gggagaatcc tggatcttat taagaaagga gtagcaaagg
     2641 ttgatcatgt ccagatgata gtattggatg aggcagataa gttgctgtca caggattttg
     2701 tgcagataat ggaggatatt attctcacgc tacctaaaaa caggcagatt ttactatatt
     2761 ccgctacttt ccctcttagt gtacagaagt tcatgaattc ccatttgcag aaaccctatg
     2821 agattaacct gatggaggaa ctaactctga agggagtaac ccagtactac gcatatgtaa
     2881 ctgagcgcca aaaagtacac tgcctcaaca cacttttctc caggcttcag ataaaccagt
     2941 cgatcatttt ctgtaactcc tctcagcgag ttgaattgct agccaagaag atttctcaac
     3001 tgggttattc ttgcttctat attcatgcta aaatgaggca ggaacatcga aatcgtgtat
     3061 ttcatgattt ccgaaatggc ttatgccgca atcttgtttg cactgatctg tttacccgag
     3121 gtattgatat acaagctgtg aatgtggtaa taaactttga tttcccaaag ctggcagaga
     3181 cctatctcca tcgtattgga agatcaggtc gctttggtca tcttggctta gccatcaact
     3241 tgatcacata tgatgatcgc ttcaacctga aaagtattga ggagcagctg ggaacagaaa
     3301 ttaaacctat tccgagcaac attgataaga gcctgtatgt ggcagaatac cacagcgagc
     3361 ctgtagaaga tgagaaacct taactcgagt ctggtaaaga aaccgctgct gcgaaatttg
     3421 aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct gctgccaccg
     3481 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     3541 tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg
     3601 taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg acgacaagct
     3661 gacgaccggg tctccgcaag tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3721 ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc
     3781 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     3841 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     3901 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     3961 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     4021 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     4081 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     4141 acgaaatacg cggtcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     4201 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     4261 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     4321 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     4381 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     4441 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     4501 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga
     4561 cgtttcccgt tgaatatggc tcatactctt cctttttcaa tattattgaa gcatttatca
     4621 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4681 catgcagcgc tcttccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
     4741 gcgagcggtg tcagctcact caaaagcggt aatacggtta tccacagaat caggggataa
     4801 agccggaaag aacatgtgag caaaaagcaa agcaccggaa gaagccaacg ccgcaggcgt
     4861 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agccagaggt
     4921 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4981 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5041 gcgtggcgct ttctcatagc tcacgctgtt ggtatctcag ttcggtgtag gtcgttcgct
     5101 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5161 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccattg
     5221 gtaactgatt tagaggactt tgtcttgaag ttatgcacct gttaaggcta aactgaaaga
     5281 acagattttg gtgagtgcgg tcctccaacc cacttacctt ggttcaaaga gttggtagct
     5341 cagcgaacct tgagaaaacc accgttggta gcggtggttt ttctttattt atgagatgat
     5401 gaatcaatcg gtctatcaag tcaacgaaca gctattccgt tactctagat ttcagtgcaa
     5461 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca
     5521 tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     5581 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     5641 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5701 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     5761 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     5821 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     5881 gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg
     5941 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     6001 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     6061 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     6121 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     6181 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     6241 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     6301 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     6361 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     6421 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     6481 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     6541 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     6601 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     6661 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     6721 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     6781 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     6841 attaatacga ctcactata
//
