LOCUS       sequence-289667-        4969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmXRN1_1323-1355-L1323AQ1326A-DmDCP1_1-127.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4969)
  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 4969)
  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_1323-1355-L1323AQ1326A-DmDCP1_1-127_P
FEATURES             Location/Qualifiers
     source          1..4969
                     /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        316..334
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    335..359
                     /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             405..1058
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1016..1038
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1065..1088
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1506..1550
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1588..1606)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1602..1649
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1879..2694)
                     /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(1879..2694)
                     /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     2602..2621
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2695..2786)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2802..3551)
                     /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(2812..2829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3052..3071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3747..4829)
                     /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     4791..4810
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4830..4907)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4953..4969,1..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 cggcacttca agcgcaaaga gctccgtttc ccggacaaat
      181 gcccaatcta cccaagccac cgctcttctg gcagcaagag gcgcagaagc aggaggcgct
      241 gtagaagctt gcggccgcat aatgcttaag tcgaacagaa agtaatcgta ttgtacacgg
      301 ccgcataatc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
      361 catcttagta tattagttaa gtataagaag gagatataca taatatgtcc cctatactag
      421 gttattggaa aattaagggc cttgtgcaac ccactcgact tcttttggaa tatcttgaag
      481 aaaaatatga agagcatttg tatgagcgcg atgaaggtga taaatggcga aacaaaaagt
      541 ttgaattggg tttggagttt cccaatcttc cttattatat tgatggtgat gttaaattaa
      601 cacagtctat ggccatcata cgttatatag ctgacaagca caacatgttg ggtggttgtc
      661 caaaagagcg tgcagagatt tcaatgcttg aaggagcggt tttggatatt agatacggtg
      721 tttcgagaat tgcatatagt aaagactttg aaactctcaa agttgatttt cttagcaagc
      781 tacctgaaat gctgaaaatg ttcgaagatc gtttatgtca taaaacatat ttaaatggtg
      841 atcatgtaac ccatcctgac ttcatgttgt atgacgctct tgatgttgtt ttatacatgg
      901 acccaatgtg cctggatgcg ttcccaaaat tagtttgttt taaaaaacgt attgaagcta
      961 tcccacaaat tgataagtac ttgaaatcca gcaagtatat agcatggcct ttgcagggct
     1021 ggcaagccac gtttggtggt ggcgaccatc ctccaaaatc ggatctggaa gttctgttcc
     1081 aggggcccca tatggcagat ctcatggccg acgagagcat cacgcgaatg aacctggcgg
     1141 ccatcaagaa gatcgacccg tacgccaagg agatcgtgga ttcgtcctcg cacgtcgcct
     1201 tctacacgtt caactcgtcg cagaacgagt gggaaaagac cgatgtggag ggagccttct
     1261 tcatatacca ccgcaacgcg gagccctttc acagcatctt catcaacaac cgactgaaca
     1321 ccacgtcctt cgtggagccc atcaccggca gcctggagct gcagtcgcag ccgccgttcc
     1381 tgctctaccg caacgagcgc tcgcgcatcc gcggcttctg gttctacaac agcgaggagt
     1441 gcgaccgcat cagcggcttg gtgaacgggc tgctcaagtc caagtagggt accctcgagt
     1501 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg
     1561 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     1621 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg
     1681 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
     1741 aacgaccctg ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt
     1801 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1861 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     1921 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     1981 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     2041 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     2101 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     2161 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     2221 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca
     2281 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     2341 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     2401 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     2461 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     2521 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     2581 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     2641 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt
     2701 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     2761 tgaatgtatt tagaaaaata aacaaatagg catgcagcgc tcttccgctt cctcgctcac
     2821 tgactcgcta cgctcggtcg ttcgactgcg gcgagcggtg tcagctcact caaaagcggt
     2881 aatacggtta tccacagaat caggggataa agccggaaag aacatgtgag caaaaagcaa
     2941 agcaccggaa gaagccaacg ccgcaggcgt ttttccatag gctccgcccc cctgacgagc
     3001 atcacaaaaa tcgacgctca agccagaggt ggcgaaaccc gacaggacta taaagatacc
     3061 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3121 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgtt
     3181 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3241 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3301 acgacttatc gccactggca gcagccattg gtaactgatt tagaggactt tgtcttgaag
     3361 ttatgcacct gttaaggcta aactgaaaga acagattttg gtgagtgcgg tcctccaacc
     3421 cacttacctt ggttcaaaga gttggtagct cagcgaacct tgagaaaacc accgttggta
     3481 gcggtggttt ttctttattt atgagatgat gaatcaatcg gtctatcaag tcaacgaaca
     3541 gctattccgt tactctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
     3601 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg
     3661 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     3721 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3781 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     3841 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     3901 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     3961 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     4021 accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     4081 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     4141 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     4201 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     4261 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     4321 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     4381 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     4441 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     4501 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     4561 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     4621 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     4681 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     4741 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     4801 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     4861 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     4921 ctctccctta tgcgactcct gcattaggaa attaatacga ctcactata
//
