LOCUS       sequence-289503-        5053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmXRN1_1323-1383-DmDCP1_1-127.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5053)
  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 5053)
  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-DmXRN1_1323-1383-DmDCP1_1-127_N
FEATURES             Location/Qualifiers
     source          1..5053
                     /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        400..418
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    419..443
                     /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             489..1142
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1100..1122
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1149..1172
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1590..1634
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1672..1690)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1686..1733
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1963..2778)
                     /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(1963..2778)
                     /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     2686..2705
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2779..2870)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2886..3635)
                     /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(2896..2913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3136..3155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3831..4913)
                     /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     4875..4894
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4914..4991)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5037..5053,1..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 cgctacttca acagcaaaga gctccgtttc ccggacaaat
      181 gcccaatcta cccaagccac cgctcttctg gcagcaagag gcgcagaagc aggaggcgct
      241 gcagcaggag gcgcagcagc aggaggcgca gaagaagcag cagcaggcgc atgcgcaaat
      301 ggagcctgaa cgtatcaata gccagtagaa gcttgcggcc gcataatgct taagtcgaac
      361 agaaagtaat cgtattgtac acggccgcat aatcgaaatt aatacgactc actatagggg
      421 aattgtgagc ggataacaat tccccatctt agtatattag ttaagtataa gaaggagata
      481 tacataatat gtcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc
      541 gacttctttt ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag
      601 gtgataaatg gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt
      661 atattgatgg tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca
      721 agcacaacat gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag
      781 cggttttgga tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc
      841 tcaaagttga ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat
      901 gtcataaaac atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg
      961 ctcttgatgt tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt
     1021 gttttaaaaa acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt
     1081 atatagcatg gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catcctccaa
     1141 aatcggatct ggaagttctg ttccaggggc cccatatggc agatctcatg gccgacgaga
     1201 gcatcacgcg aatgaacctg gcggccatca agaagatcga cccgtacgcc aaggagatcg
     1261 tggattcgtc ctcgcacgtc gccttctaca cgttcaactc gtcgcagaac gagtgggaaa
     1321 agaccgatgt ggagggagcc ttcttcatat accaccgcaa cgcggagccc tttcacagca
     1381 tcttcatcaa caaccgactg aacaccacgt ccttcgtgga gcccatcacc ggcagcctgg
     1441 agctgcagtc gcagccgccg ttcctgctct accgcaacga gcgctcgcgc atccgcggct
     1501 tctggttcta caacagcgag gagtgcgacc gcatcagcgg cttggtgaac gggctgctca
     1561 agtccaagta gggtaccctc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc
     1621 agcacatgga ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc
     1681 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac
     1741 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
     1801 agcaatagac ataagcggct atttaacgac cctgccctga accgacgaca agctgacgac
     1861 cgggtctccg caagtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1921 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     1981 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     2041 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     2101 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     2161 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     2221 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     2281 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     2341 tacgcggtcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     2401 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     2461 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     2521 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     2581 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     2641 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     2701 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     2761 ccgttgaata tggctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2821 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggcatgca
     2881 gcgctcttcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     2941 ggtgtcagct cactcaaaag cggtaatacg gttatccaca gaatcagggg ataaagccgg
     3001 aaagaacatg tgagcaaaaa gcaaagcacc ggaagaagcc aacgccgcag gcgtttttcc
     3061 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagccag aggtggcgaa
     3121 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3181 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3241 cgctttctca tagctcacgc tgttggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3301 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3361 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc attggtaact
     3421 gatttagagg actttgtctt gaagttatgc acctgttaag gctaaactga aagaacagat
     3481 tttggtgagt gcggtcctcc aacccactta ccttggttca aagagttggt agctcagcga
     3541 accttgagaa aaccaccgtt ggtagcggtg gtttttcttt atttatgaga tgatgaatca
     3601 atcggtctat caagtcaacg aacagctatt ccgttactct agatttcagt gcaatttatc
     3661 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag
     3721 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     3781 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     3841 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3901 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     3961 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     4021 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     4081 tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg
     4141 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     4201 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     4261 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     4321 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     4381 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     4441 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     4501 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     4561 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     4621 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     4681 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     4741 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     4801 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     4861 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     4921 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     4981 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaaattaat
     5041 acgactcact ata
//
