LOCUS       sequence-289256-        5827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsEDC4_977-1401-HsDCP2_385-420.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5827)
  AUTHORS   Chang CT, Bercovich N, Loh B, Jonas S, Izaurralde E
  TITLE     The activation of the decapping enzyme DCP2 by DCP1 occurs on the 
            EDC4 scaffold and involves a conserved loop in DCP1.
  JOURNAL   Nucleic Acids Res. 2014 Apr;42(8):5217-33. doi: 10.1093/nar/gku129. 
            Epub 2014 Feb 8.
  PUBMED    24510189
REFERENCE   2  (bases 1 to 5827)
  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: 
            "10.1093/nar/gku129"; journalName: "Nucleic Acids Res"; date: 
            "2014-04"; volume: "42"; issue: "8"; pages: "5217-33"
COMMENT     Sequence Label: pRSFDuetG-HsEDC4_977-1401-HsDCP2_395-420_K
FEATURES             Location/Qualifiers
     source          1..5827
                     /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        1486..1504
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1505..1529
                     /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             1575..2228
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2186..2208
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2235..2258
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2364..2408
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2446..2464)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2460..2507
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2737..3552)
                     /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(2737..3552)
                     /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     3460..3479
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3553..3644)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3660..4409)
                     /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(3670..3687)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3910..3929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4605..5687)
                     /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     5649..5668
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5688..5765)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5811..5827,1..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 cgacccaact cgaaggcctg cagagcacag tcacaggcca
      181 cgtagaacgt gcccttgaga ctcggcacga gcaggaacag cggcggctgg agcgagcact
      241 ggctgagggg cagcagcggg gagggcagct gcaggagcag ctgacacaac agttgtccca
      301 agcactgtcg tcagctgtag ctgggcggct agagcgcagc atacgggatg agatcaagaa
      361 gacagtccct ccatgtgtct caaggagtct ggagcctatg gcaggccaac tgagcaactc
      421 agtggctacc aagctcacag ctgtggaggg cagcatgaaa gagaacatct ccaagctgct
      481 caagtccaag aacttgactg atgccatcgc ccgagcagct gcagacacat tacaagggcc
      541 gatgcaggct gcctaccggg aagccttcca gagtgtggtg ctgccggcct ttgagaagag
      601 ctgccaggcc atgttccagc aaatcaatga tagcttccgg ctggggacac aggaatactt
      661 gcagcagcta gaaagccaca tgaagagccg gaaggcacgg gaacaggagg ccagggagcc
      721 tgtgctagcc cagctgcggg gcctggtcag cacactgcag agtgccactg agcagatggc
      781 agccaccgtg gccggcagtg ttcgtgctga ggtgcagcac cagctgcatg tggctgtggg
      841 cagcctgcag gagtccattt tagcacaggt acagcgcatc gttaagggtg aggtgagtgt
      901 ggcgctcaag gagcagcagg ccgccgtcac ctccagcatc atgcaggcca tgcgctcagc
      961 tgctggcaca cctgtcccct ctgcccacct tgactgccag gcccagcaag cccatatcct
     1021 gcagctgctg cagcagggcc acctcaatca ggccttccag caggcgctga cagctgctga
     1081 cctgaacctg gtgctgtatg tgtgtgaaac tgtggaccca gcccaggttt ttgggcagcc
     1141 accctgcccg ctctcccagc ctgtgctcct ttccctcatc cagcagctgg catctgacct
     1201 tggcactcga actgacctca agctcagcta cctggaagag gccgtgatgc acctggacca
     1261 cagtgacccc atcactcggg accacatggg ctccgttatg gcccaggtgc gccaaaagct
     1321 ttttcagttc ctgcaggctg agccacacaa ctcacttggc aaagcagctc ggcgtctcag
     1381 cctcatgctg catggcctcg tgacccccag cctcccttaa gcggccgcat aatgcttaag
     1441 tcgaacagaa agtaatcgta ttgtacacgg ccgcataatc gaaattaata cgactcacta
     1501 taggggaatt gtgagcggat aacaattccc catcttagta tattagttaa gtataagaag
     1561 gagatataca taatatgtcc cctatactag gttattggaa aattaagggc cttgtgcaac
     1621 ccactcgact tcttttggaa tatcttgaag aaaaatatga agagcatttg tatgagcgcg
     1681 atgaaggtga taaatggcga aacaaaaagt ttgaattggg tttggagttt cccaatcttc
     1741 cttattatat tgatggtgat gttaaattaa cacagtctat ggccatcata cgttatatag
     1801 ctgacaagca caacatgttg ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg
     1861 aaggagcggt tttggatatt agatacggtg tttcgagaat tgcatatagt aaagactttg
     1921 aaactctcaa agttgatttt cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc
     1981 gtttatgtca taaaacatat ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt
     2041 atgacgctct tgatgttgtt ttatacatgg acccaatgtg cctggatgcg ttcccaaaat
     2101 tagtttgttt taaaaaacgt attgaagcta tcccacaaat tgataagtac ttgaaatcca
     2161 gcaagtatat agcatggcct ttgcagggct ggcaagccac gtttggtggt ggcgaccatc
     2221 ctccaaaatc ggatctggaa gttctgttcc aggggcccca tatgtgcaag ttcccctttt
     2281 catccagagc ctttttgagt ttcaagtttg accataatgc tataatgaaa atcttggacc
     2341 tttaaggtac cctcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca
     2401 tggactcgtc tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac
     2461 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg
     2521 catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat
     2581 agacataagc ggctatttaa cgaccctgcc ctgaaccgac gacaagctga cgaccgggtc
     2641 tccgcaagtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2701 atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa
     2761 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     2821 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     2881 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     2941 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     3001 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     3061 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     3121 gtcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     3181 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     3241 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     3301 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     3361 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     3421 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     3481 atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg
     3541 aatatggctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3601 catgagcgga tacatatttg aatgtattta gaaaaataaa caaataggca tgcagcgctc
     3661 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggtgtc
     3721 agctcactca aaagcggtaa tacggttatc cacagaatca ggggataaag ccggaaagaa
     3781 catgtgagca aaaagcaaag caccggaaga agccaacgcc gcaggcgttt ttccataggc
     3841 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag ccagaggtgg cgaaacccga
     3901 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3961 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4021 ctcatagctc acgctgttgg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4081 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4141 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccattggt aactgattta
     4201 gaggactttg tcttgaagtt atgcacctgt taaggctaaa ctgaaagaac agattttggt
     4261 gagtgcggtc ctccaaccca cttaccttgg ttcaaagagt tggtagctca gcgaaccttg
     4321 agaaaaccac cgttggtagc ggtggttttt ctttatttat gagatgatga atcaatcggt
     4381 ctatcaagtc aacgaacagc tattccgtta ctctagattt cagtgcaatt tatctcttca
     4441 aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc
     4501 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
     4561 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     4621 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4681 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     4741 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     4801 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     4861 gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg
     4921 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     4981 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     5041 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     5101 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     5161 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     5221 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     5281 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     5341 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     5401 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     5461 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     5521 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     5581 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     5641 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     5701 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     5761 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact
     5821 cactata
//
