LOCUS       sequence-288854-        4648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsDCP1a_539-582-mut2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4648)
  AUTHORS   Tritschler F, Braun JE, Motz C, Igreja C, Haas G, Truffault V, 
            Izaurralde E, Weichenrieder O
  TITLE     DCP1 forms asymmetric trimers to assemble into active mRNA decapping
            complexes in metazoa.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Dec 22. 106(51):21591-6.
  PUBMED    19966221
REFERENCE   2  (bases 1 to 4648)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2009 Dec 22. 106(51):21591-6."
COMMENT     Sequence Label: pRSFDuetG-HsDCP1a_539-582 (MCSII)-mut2_G
FEATURES             Location/Qualifiers
     source          1..4648
                     /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        244..262
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    263..287
                     /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             333..986
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     944..966
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             993..1016
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1185..1229
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1267..1285)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1281..1328
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1558..2373)
                     /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(1558..2373)
                     /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     2281..2300
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2374..2465)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2481..3230)
                     /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(2491..2508)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2731..2750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3426..4508)
                     /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     4470..4489
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4509..4586)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4632..4648,1..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 cgaattcgag ctcggcgcgc ctgcaggtcg acaagcttgc
      181 ggccgcataa tgcttaagtc gaacagaaag taatcgtatt gtacacggcc gcataatcga
      241 aattaatacg actcactata ggggaattgt gagcggataa caattcccca tcttagtata
      301 ttagttaagt ataagaagga gatatacata atatgtcccc tatactaggt tattggaaaa
      361 ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag
      421 agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt
      481 tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg
      541 ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg
      601 cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg
      661 catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc
      721 tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc
      781 atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc
      841 tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg
      901 ataagtactt gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt
      961 ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt tctgttccag gggcccagtc
     1021 agagaaagcc ttccattatt ctcagcaagt ctcagctcca ggatacatta tcccatctac
     1081 gcaagaatga ttccagcttc gaaagtacac ttcatgaagt ctacttgcag gtgctgacca
     1141 agaacaaaga caaccacaac ctatgaggta ccctcgagtc tggtaaagaa accgctgctg
     1201 cgaaatttga acgccagcac atggactcgt ctactagcgc agcttaatta acctaggctg
     1261 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     1321 gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca
     1381 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga
     1441 cgacaagctg acgaccgggt ctccgcaagt ggcacttttc ggggaaatgt gcgcggaacc
     1501 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta
     1561 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     1621 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     1681 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     1741 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     1801 atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc
     1861 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     1921 ctgagcgaga cgaaatacgc ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     1981 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     2041 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     2101 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     2161 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     2221 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     2281 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     2341 agagcaagac gtttcccgtt gaatatggct catactcttc ctttttcaat attattgaag
     2401 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     2461 acaaataggc atgcagcgct cttccgcttc ctcgctcact gactcgctac gctcggtcgt
     2521 tcgactgcgg cgagcggtgt cagctcactc aaaagcggta atacggttat ccacagaatc
     2581 aggggataaa gccggaaaga acatgtgagc aaaaagcaaa gcaccggaag aagccaacgc
     2641 cgcaggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2701 gccagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2761 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2821 cttcgggaag cgtggcgctt tctcatagct cacgctgttg gtatctcagt tcggtgtagg
     2881 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2941 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3001 cagccattgg taactgattt agaggacttt gtcttgaagt tatgcacctg ttaaggctaa
     3061 actgaaagaa cagattttgg tgagtgcggt cctccaaccc acttaccttg gttcaaagag
     3121 ttggtagctc agcgaacctt gagaaaacca ccgttggtag cggtggtttt tctttattta
     3181 tgagatgatg aatcaatcgg tctatcaagt caacgaacag ctattccgtt actctagatt
     3241 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     3301 taattctcat gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     3361 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     3421 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3481 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     3541 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     3601 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     3661 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg
     3721 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     3781 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     3841 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     3901 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     3961 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     4021 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     4081 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     4141 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     4201 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     4261 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     4321 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     4381 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     4441 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     4501 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     4561 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     4621 cattaggaaa ttaatacgac tcactata
//
