LOCUS       sequence-360752-        5242 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial Expression of DmDcp1_128-372.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5242)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5242)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pRSFDuetG-DmDCP1_128-372 (MCSI)_E
FEATURES             Location/Qualifiers
     source          1..5242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..150
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        954..972
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    973..997
                     /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             1043..1696
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1654..1676
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1703..1726
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1796..1840
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1878..1896)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1892..1939
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2169..2984)
                     /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(2169..2984)
                     /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     2892..2911
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2985..3076)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3092..3841)
                     /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(3102..3119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3342..3361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4037..5119)
                     /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     5081..5100
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5120..5197)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcaccatca tcaccacagc
      121 gcgggtctgg aagttctgtt ccaggggccc caggatccgg atcagggaac gaatggccag
      181 gcccagcgtc acgtctccgc gccccagcag ccaaagcagg acagcagcca gccggccagc
      241 atattcaaca tgctgaccaa ggcccagaag gactacaatg cccaggtgag cggcgggcag
      301 ccgaagacgc cttcggcgga gaacgtaacg gccggcaatg tgttgaagtt cttcgaatcg
      361 gccaagcagg ccacggcaga gtcactgttc caccgcgtcc agccgctctc cgtggaccag
      421 ctggagaaac agcagcgggc agccacgcct ggcgaggatc tgttgccccc ggcaggcagg
      481 gacaatcacg agagccggct gtcgcccttc cagaagatgc agcagcttaa cattcagctc
      541 ctgagcgacc tgaagatcgg taaaagctac gctacggctc aatctccgac agtcgccccg
      601 tcctcgtcgg cgctgatggc caacgacgat gccggcaagt gcttgcggag actgctggct
      661 ggcgacaagc ccgggccggc cctcatgccg cccaccatgt tcgacgcgcc caacaacggc
      721 aatccggagc cacagcagcc acagcagcca ctgctgaact cgacgcagtt cgtccaggcc
      781 ttcacgtacc ttatccagaa cgataaggag tttgccaaca agctgcacaa ggcctacctg
      841 aacggctgtt ccaatctact gctggactcc agctccacat atcaataagc ggccgcataa
      901 tgcttaagtc gaacagaaag taatcgtatt gtacacggcc gcataatcga aattaatacg
      961 actcactata ggggaattgt gagcggataa caattcccca tcttagtata ttagttaagt
     1021 ataagaagga gatatacata atatgtcccc tatactaggt tattggaaaa ttaagggcct
     1081 tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta
     1141 tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc
     1201 caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg
     1261 ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc
     1321 aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa
     1381 agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt
     1441 cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt
     1501 catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt
     1561 cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt
     1621 gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg
     1681 cgaccatcct ccaaaatcgg atctggaagt tctgttccag gggccccata tggcagatct
     1741 caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt ctggtaaaga
     1801 aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg cagcttaatt
     1861 aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1921 ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct
     1981 tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg
     2041 ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt cggggaaatg
     2101 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2161 attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     2221 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     2281 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     2341 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     2401 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     2461 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     2521 cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca attacaaaca
     2581 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     2641 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     2701 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     2761 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     2821 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     2881 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     2941 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt cctttttcaa
     3001 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3061 tagaaaaata aacaaatagg catgcagcgc tcttccgctt cctcgctcac tgactcgcta
     3121 cgctcggtcg ttcgactgcg gcgagcggtg tcagctcact caaaagcggt aatacggtta
     3181 tccacagaat caggggataa agccggaaag aacatgtgag caaaaagcaa agcaccggaa
     3241 gaagccaacg ccgcaggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3301 tcgacgctca agccagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3361 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3421 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgtt ggtatctcag
     3481 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3541 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3601 gccactggca gcagccattg gtaactgatt tagaggactt tgtcttgaag ttatgcacct
     3661 gttaaggcta aactgaaaga acagattttg gtgagtgcgg tcctccaacc cacttacctt
     3721 ggttcaaaga gttggtagct cagcgaacct tgagaaaacc accgttggta gcggtggttt
     3781 ttctttattt atgagatgat gaatcaatcg gtctatcaag tcaacgaaca gctattccgt
     3841 tactctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac
     3901 gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg
     3961 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     4021 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4081 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     4141 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     4201 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     4261 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt
     4321 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     4381 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     4441 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     4501 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     4561 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     4621 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     4681 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     4741 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     4801 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     4861 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     4921 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     4981 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     5041 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     5101 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     5161 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     5221 tgcgactcct gcattaggaa at
//
