LOCUS       sequence-288850-        4633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmDCP1_328-366-mut3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4633)
  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 4633)
  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-DmDCP1_328-366 (MCSII)-mut3_G
FEATURES             Location/Qualifiers
     source          1..4633
                     /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             1170..1214
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1252..1270)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1266..1313
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1543..2358)
                     /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(1543..2358)
                     /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     2266..2285
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2359..2450)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2466..3215)
                     /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(2476..2493)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2716..2735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3411..4493)
                     /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     4455..4474
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4494..4571)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4617..4633,1..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 gggccccagc
     1021 cacagcagcc actgctgaac tcgacgcagt tcgtccaggc ccgcacgtac ctttcccaga
     1081 acgataagga gcgcgccaac aagtcccaca aggcctacct gaacggctgt tccaatctac
     1141 tgctggacta gggtaccctc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc
     1201 agcacatgga ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc
     1261 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac
     1321 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
     1381 agcaatagac ataagcggct atttaacgac cctgccctga accgacgaca agctgacgac
     1441 cgggtctccg caagtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1501 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     1561 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     1621 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     1681 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     1741 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     1801 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     1861 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     1921 tacgcggtcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     1981 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     2041 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     2101 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     2161 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     2221 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     2281 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     2341 ccgttgaata tggctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2401 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggcatgca
     2461 gcgctcttcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     2521 ggtgtcagct cactcaaaag cggtaatacg gttatccaca gaatcagggg ataaagccgg
     2581 aaagaacatg tgagcaaaaa gcaaagcacc ggaagaagcc aacgccgcag gcgtttttcc
     2641 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagccag aggtggcgaa
     2701 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2761 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2821 cgctttctca tagctcacgc tgttggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2881 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2941 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc attggtaact
     3001 gatttagagg actttgtctt gaagttatgc acctgttaag gctaaactga aagaacagat
     3061 tttggtgagt gcggtcctcc aacccactta ccttggttca aagagttggt agctcagcga
     3121 accttgagaa aaccaccgtt ggtagcggtg gtttttcttt atttatgaga tgatgaatca
     3181 atcggtctat caagtcaacg aacagctatt ccgttactct agatttcagt gcaatttatc
     3241 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag
     3301 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     3361 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     3421 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3481 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     3541 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     3601 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     3661 tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg
     3721 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     3781 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     3841 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     3901 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     3961 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     4021 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     4081 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     4141 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     4201 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     4261 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     4321 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     4381 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     4441 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     4501 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     4561 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaaattaat
     4621 acgactcact ata
//
