LOCUS       sequence-289468-        5077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmDCP1_1-127-DmXRN1_1312-1383.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5077)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5077)
  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"
COMMENT     Sequence Label: pRSFDuetG-DmDCP1_1-127-DmXRN1_1312-1383_N
FEATURES             Location/Qualifiers
     source          1..5077
                     /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        598..616
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    617..641
                     /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             687..1340
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1298..1320
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1347..1370
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1614..1658
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1696..1714)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1710..1757
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1987..2802)
                     /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(1987..2802)
                     /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     2710..2729
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2803..2894)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2910..3659)
                     /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(2920..2937)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3160..3179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3855..4937)
                     /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     4899..4918
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4938..5015)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5061..5077,1..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 cgatggccga cgagagcatc acgcgaatga acctggcggc
      181 catcaagaag atcgacccgt acgccaagga gatcgtggat tcgtcctcgc acgtcgcctt
      241 ctacacgttc aactcgtcgc agaacgagtg ggaaaagacc gatgtggagg gagccttctt
      301 catataccac cgcaacgcgg agccctttca cagcatcttc atcaacaacc gactgaacac
      361 cacgtccttc gtggagccca tcaccggcag cctggagctg cagtcgcagc cgccgttcct
      421 gctctaccgc aacgagcgct cgcgcatccg cggcttctgg ttctacaaca gcgaggagtg
      481 cgaccgcatc agcggcttgg tgaacgggct gctcaagtcc aagtagaagc ttgcggccgc
      541 ataatgctta agtcgaacag aaagtaatcg tattgtacac ggccgcataa tcgaaattaa
      601 tacgactcac tataggggaa ttgtgagcgg ataacaattc cccatcttag tatattagtt
      661 aagtataaga aggagatata cataatatgt cccctatact aggttattgg aaaattaagg
      721 gccttgtgca acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt
      781 tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt
      841 ttcccaatct tccttattat attgatggtg atgttaaatt aacacagtct atggccatca
      901 tacgttatat agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga
      961 tttcaatgct tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata
     1021 gtaaagactt tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa
     1081 tgttcgaaga tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg
     1141 acttcatgtt gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg
     1201 cgttcccaaa attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt
     1261 acttgaaatc cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg
     1321 gtggcgacca tcctccaaaa tcggatctgg aagttctgtt ccaggggccc catatgctcc
     1381 aggaggcggt gcagcagcaa cgtccgccac tacttcaaca gcaaagagct ccgtttcccg
     1441 gacaaatgcc caatctaccc aagccaccgc tcttctggca gcaagaggcg cagaagcagg
     1501 aggcgctgca gcaggaggcg cagcagcagg aggcgcagaa gaagcagcag caggcgcatg
     1561 cgcaaatgga gcctgaacgt atcaatagcc agtagggtac cctcgagtct ggtaaagaaa
     1621 ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc tactagcgca gcttaattaa
     1681 cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1741 tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc
     1801 cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc
     1861 ctgaaccgac gacaagctga cgaccgggtc tccgcaagtg gcacttttcg gggaaatgtg
     1921 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat
     1981 taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     2041 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     2101 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     2161 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     2221 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     2281 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     2341 tgattgcgcc tgagcgagac gaaatacgcg gtcgctgtta aaaggacaat tacaaacagg
     2401 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     2461 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     2521 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     2581 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     2641 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     2701 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     2761 tcgcggccta gagcaagacg tttcccgttg aatatggctc atactcttcc tttttcaata
     2821 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2881 gaaaaataaa caaataggca tgcagcgctc ttccgcttcc tcgctcactg actcgctacg
     2941 ctcggtcgtt cgactgcggc gagcggtgtc agctcactca aaagcggtaa tacggttatc
     3001 cacagaatca ggggataaag ccggaaagaa catgtgagca aaaagcaaag caccggaaga
     3061 agccaacgcc gcaggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3121 gacgctcaag ccagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3181 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3241 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgttgg tatctcagtt
     3301 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3361 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3421 cactggcagc agccattggt aactgattta gaggactttg tcttgaagtt atgcacctgt
     3481 taaggctaaa ctgaaagaac agattttggt gagtgcggtc ctccaaccca cttaccttgg
     3541 ttcaaagagt tggtagctca gcgaaccttg agaaaaccac cgttggtagc ggtggttttt
     3601 ctttatttat gagatgatga atcaatcggt ctatcaagtc aacgaacagc tattccgtta
     3661 ctctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
     3721 tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag ctgactgggt
     3781 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     3841 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3901 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     3961 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     4021 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     4081 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc
     4141 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     4201 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     4261 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     4321 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     4381 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     4441 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     4501 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     4561 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     4621 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     4681 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     4741 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     4801 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     4861 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     4921 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     4981 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     5041 cgactcctgc attaggaaat taatacgact cactata
//
