LOCUS       sequence-289501-        5002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmXRN1_1312-1355-DmDCP1_1-127.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5002)
  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 5002)
  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_1312-1355-DmDCP1_1-127_N
FEATURES             Location/Qualifiers
     source          1..5002
                     /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        349..367
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    368..392
                     /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             438..1091
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1049..1071
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1098..1121
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1539..1583
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1621..1639)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1635..1682
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1912..2727)
                     /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(1912..2727)
                     /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     2635..2654
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2728..2819)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2835..3584)
                     /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(2845..2862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3085..3104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3780..4862)
                     /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     4824..4843
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4863..4940)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4986..5002,1..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 cgctccagga ggcggtgcag cagcaacgtc cgccactact
      181 tcaacagcaa agagctccgt ttcccggaca aatgcccaat ctacccaagc caccgctctt
      241 ctggcagcaa gaggcgcaga agcaggaggc gctgtagaag cttgcggccg cataatgctt
      301 aagtcgaaca gaaagtaatc gtattgtaca cggccgcata atcgaaatta atacgactca
      361 ctatagggga attgtgagcg gataacaatt ccccatctta gtatattagt taagtataag
      421 aaggagatat acataatatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      481 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      541 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      601 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      661 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      721 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      781 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      841 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      901 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      961 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
     1021 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
     1081 atcctccaaa atcggatctg gaagttctgt tccaggggcc ccatatggca gatctcatgg
     1141 ccgacgagag catcacgcga atgaacctgg cggccatcaa gaagatcgac ccgtacgcca
     1201 aggagatcgt ggattcgtcc tcgcacgtcg ccttctacac gttcaactcg tcgcagaacg
     1261 agtgggaaaa gaccgatgtg gagggagcct tcttcatata ccaccgcaac gcggagccct
     1321 ttcacagcat cttcatcaac aaccgactga acaccacgtc cttcgtggag cccatcaccg
     1381 gcagcctgga gctgcagtcg cagccgccgt tcctgctcta ccgcaacgag cgctcgcgca
     1441 tccgcggctt ctggttctac aacagcgagg agtgcgaccg catcagcggc ttggtgaacg
     1501 ggctgctcaa gtccaagtag ggtaccctcg agtctggtaa agaaaccgct gctgcgaaat
     1561 ttgaacgcca gcacatggac tcgtctacta gcgcagctta attaacctag gctgctgcca
     1621 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1681 tgctgaaacc tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac
     1741 cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgacaa
     1801 gctgacgacc gggtctccgc aagtggcact tttcggggaa atgtgcgcgg aacccctatt
     1861 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa
     1921 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     1981 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     2041 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     2101 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     2161 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     2221 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     2281 gagacgaaat acgcggtcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     2341 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     2401 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     2461 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     2521 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     2581 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     2641 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca
     2701 agacgtttcc cgttgaatat ggctcatact cttccttttt caatattatt gaagcattta
     2761 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2821 aggcatgcag cgctcttccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact
     2881 gcggcgagcg gtgtcagctc actcaaaagc ggtaatacgg ttatccacag aatcagggga
     2941 taaagccgga aagaacatgt gagcaaaaag caaagcaccg gaagaagcca acgccgcagg
     3001 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagccaga
     3061 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3121 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3181 gaagcgtggc gctttctcat agctcacgct gttggtatct cagttcggtg taggtcgttc
     3241 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3301 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3361 ttggtaactg atttagagga ctttgtcttg aagttatgca cctgttaagg ctaaactgaa
     3421 agaacagatt ttggtgagtg cggtcctcca acccacttac cttggttcaa agagttggta
     3481 gctcagcgaa ccttgagaaa accaccgttg gtagcggtgg tttttcttta tttatgagat
     3541 gatgaatcaa tcggtctatc aagtcaacga acagctattc cgttactcta gatttcagtg
     3601 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc
     3661 tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     3721 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     3781 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3841 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     3901 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     3961 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     4021 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     4081 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     4141 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     4201 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     4261 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     4321 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     4381 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     4441 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     4501 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     4561 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     4621 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     4681 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     4741 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     4801 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     4861 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     4921 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     4981 gaaattaata cgactcacta ta
//
