LOCUS       sequence-288658-        6019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsEDC3_192-228-HsRck-K135A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6019)
  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 6019)
  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_192-228-GST-HsRCK-K135A_E
FEATURES             Location/Qualifiers
     source          1..6019
                     /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        328..346
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    347..371
                     /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             432..1085
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1043..1065
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1092..1115
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2556..2600
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2638..2656)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2652..2699
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2929..3744)
                     /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(2929..3744)
                     /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     3652..3671
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3745..3836)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3852..4601)
                     /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(3862..3879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4102..4121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4797..5879)
                     /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     5841..5860
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5880..5957)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6003..6019,1..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 gaaaggagat aggcggccgc ataatgctta agtcgaacag aaagtaatcg
      301 tattgtacac ggccgcataa tcgaaattaa tacgactcac tataggggaa ttgtgagcgg
      361 ataacaattc cccatcttag tatattagtt aagtataaga aggagatata catatggcag
      421 atctcaattg gatgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca
      481 ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg
      541 aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt
      601 attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg
      661 acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag
      721 gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa
      781 ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt
      841 tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg
      901 acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag
      961 tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca
     1021 agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc
     1081 caaaatcgga tctggaagtt ctgttccagg ggcccctggg atccatgggt ctgtccagtc
     1141 aaaatggtca gctgagaggc cctgtgaaac ccactggtgg ccctggagga gggggcacac
     1201 agacacagca acagatgaac cagctgaaaa acaccaacac aatcaataat ggcactcagc
     1261 agcaagcaca gagtatgacc accactatta aacctggtga tgactggaaa aagactttaa
     1321 aactccctcc aaaggatcta agaatcaaaa cttcggatgt gacctccaca aaaggaaatg
     1381 agtttgaaga ttactgtttg aaacgggagt tactgatggg aatttttgaa atgggctggg
     1441 aaaagccatc tcctattcag gaggagagca ttcccattgc tttatctggt agggatatct
     1501 tagctagagc aaaaaatgga acaggcgcaa gcggtgccta cctcattccc ttacttgaac
     1561 ggctagacct gaagaaggac aatatacaag caatggtgat tgttcccact agagaacttg
     1621 ctctacaggt cagtcaaatt tgcatccagg tcagcaaaca catgggaggg gccaaagtga
     1681 tggcaaccac aggaggaacc aatttacgag atgacataat gaggcttgat gatacagtgc
     1741 acgtggtgat tgctacccct gggagaatcc tggatcttat taagaaagga gtagcaaagg
     1801 ttgatcatgt ccagatgata gtattggatg aggcagataa gttgctgtca caggattttg
     1861 tgcagataat ggaggatatt attctcacgc tacctaaaaa caggcagatt ttactatatt
     1921 ccgctacttt ccctcttagt gtacagaagt tcatgaattc ccatttgcag aaaccctatg
     1981 agattaacct gatggaggaa ctaactctga agggagtaac ccagtactac gcatatgtaa
     2041 ctgagcgcca aaaagtacac tgcctcaaca cacttttctc caggcttcag ataaaccagt
     2101 cgatcatttt ctgtaactcc tctcagcgag ttgaattgct agccaagaag atttctcaac
     2161 tgggttattc ttgcttctat attcatgcta aaatgaggca ggaacatcga aatcgtgtat
     2221 ttcatgattt ccgaaatggc ttatgccgca atcttgtttg cactgatctg tttacccgag
     2281 gtattgatat acaagctgtg aatgtggtaa taaactttga tttcccaaag ctggcagaga
     2341 cctatctcca tcgtattgga agatcaggtc gctttggtca tcttggctta gccatcaact
     2401 tgatcacata tgatgatcgc ttcaacctga aaagtattga ggagcagctg ggaacagaaa
     2461 ttaaacctat tccgagcaac attgataaga gcctgtatgt ggcagaatac cacagcgagc
     2521 ctgtagaaga tgagaaacct taactcgagt ctggtaaaga aaccgctgct gcgaaatttg
     2581 aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct gctgccaccg
     2641 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     2701 tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg
     2761 taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg acgacaagct
     2821 gacgaccggg tctccgcaag tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2881 ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc
     2941 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     3001 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     3061 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     3121 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     3181 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     3241 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     3301 acgaaatacg cggtcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     3361 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     3421 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     3481 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     3541 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     3601 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     3661 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga
     3721 cgtttcccgt tgaatatggc tcatactctt cctttttcaa tattattgaa gcatttatca
     3781 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3841 catgcagcgc tcttccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
     3901 gcgagcggtg tcagctcact caaaagcggt aatacggtta tccacagaat caggggataa
     3961 agccggaaag aacatgtgag caaaaagcaa agcaccggaa gaagccaacg ccgcaggcgt
     4021 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agccagaggt
     4081 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4141 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4201 gcgtggcgct ttctcatagc tcacgctgtt ggtatctcag ttcggtgtag gtcgttcgct
     4261 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4321 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccattg
     4381 gtaactgatt tagaggactt tgtcttgaag ttatgcacct gttaaggcta aactgaaaga
     4441 acagattttg gtgagtgcgg tcctccaacc cacttacctt ggttcaaaga gttggtagct
     4501 cagcgaacct tgagaaaacc accgttggta gcggtggttt ttctttattt atgagatgat
     4561 gaatcaatcg gtctatcaag tcaacgaaca gctattccgt tactctagat ttcagtgcaa
     4621 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca
     4681 tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     4741 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     4801 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4861 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     4921 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     4981 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     5041 gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg
     5101 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     5161 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     5221 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     5281 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     5341 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     5401 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     5461 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     5521 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     5581 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     5641 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     5701 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     5761 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     5821 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     5881 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     5941 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     6001 attaatacga ctcactata
//
