LOCUS       sequence-289229-        5791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsEDC4_977-1401.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5791)
  AUTHORS   Braun JE, Truffault V, Boland A, Huntzinger E, Chang CT, Haas G, 
            Weichenrieder O, Coles M, Izaurralde E
  TITLE     A direct interaction between DCP1 and XRN1 couples mRNA decapping to
            5' exonucleolytic degradation.
  JOURNAL   Nat Struct Mol Biol. 2012 Dec;19(12):1324-31. doi: 
            10.1038/nsmb.2413. Epub 2012 Nov 11.
  PUBMED    23142987
REFERENCE   2  (bases 1 to 5791)
  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: "10.1038/nsmb.2413";
            journalName: "Nat Struct Mol Biol"; date: "2012-12"; volume: "19"; 
            issue: "12"; pages: "1324-31"
COMMENT     Sequence Label: pRSFDuetG-HsEDC4_977-1401_K
FEATURES             Location/Qualifiers
     source          1..5791
                     /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             2328..2372
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2410..2428)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2424..2471
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2701..3516)
                     /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(2701..3516)
                     /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     3424..3443
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3517..3608)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3624..4373)
                     /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(3634..3651)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3874..3893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4569..5651)
                     /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     5613..5632
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5652..5729)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5775..5791,1..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 tatggcagat ctcaattgga
     2281 tatcggccgg ccacgcgatc gctgacgtcg gtaccctcga gtctggtaaa gaaaccgctg
     2341 ctgcgaaatt tgaacgccag cacatggact cgtctactag cgcagcttaa ttaacctagg
     2401 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     2461 ggggtttttt gctgaaacct caggcatttg agaagcacac ggtcacactg cttccggtag
     2521 tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac
     2581 cgacgacaag ctgacgaccg ggtctccgca agtggcactt ttcggggaaa tgtgcgcgga
     2641 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc
     2701 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     2761 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     2821 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     2881 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     2941 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     3001 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     3061 cgcctgagcg agacgaaata cgcggtcgct gttaaaagga caattacaaa caggaatcga
     3121 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     3181 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     3241 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     3301 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     3361 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     3421 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     3481 cctagagcaa gacgtttccc gttgaatatg gctcatactc ttcctttttc aatattattg
     3541 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3601 taaacaaata ggcatgcagc gctcttccgc ttcctcgctc actgactcgc tacgctcggt
     3661 cgttcgactg cggcgagcgg tgtcagctca ctcaaaagcg gtaatacggt tatccacaga
     3721 atcaggggat aaagccggaa agaacatgtg agcaaaaagc aaagcaccgg aagaagccaa
     3781 cgccgcaggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3841 caagccagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3901 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3961 tcccttcggg aagcgtggcg ctttctcata gctcacgctg ttggtatctc agttcggtgt
     4021 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4081 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4141 cagcagccat tggtaactga tttagaggac tttgtcttga agttatgcac ctgttaaggc
     4201 taaactgaaa gaacagattt tggtgagtgc ggtcctccaa cccacttacc ttggttcaaa
     4261 gagttggtag ctcagcgaac cttgagaaaa ccaccgttgg tagcggtggt ttttctttat
     4321 ttatgagatg atgaatcaat cggtctatca agtcaacgaa cagctattcc gttactctag
     4381 atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag
     4441 ttgtaattct catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     4501 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     4561 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4621 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     4681 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     4741 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     4801 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca
     4861 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     4921 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     4981 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     5041 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     5101 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     5161 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     5221 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     5281 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     5341 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     5401 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     5461 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     5521 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     5581 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     5641 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     5701 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     5761 ctgcattagg aaattaatac gactcactat a
//
