LOCUS       sequence-288621-        6235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmXRN1_1-573.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6235)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6235)
  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-DmXRN1_1-573 (MCSI)_E
FEATURES             Location/Qualifiers
     source          1..6235
                     /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        1930..1948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1949..1973
                     /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             2019..2672
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2630..2652
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2679..2702
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2772..2816
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2854..2872)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2868..2915
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3145..3960)
                     /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(3145..3960)
                     /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     3868..3887
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3961..4052)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4068..4817)
                     /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(4078..4095)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4318..4337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5013..6095)
                     /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     6057..6076
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6096..6173)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6219..6235,1..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 tgatgggcgt tcccaagttc tttcgctaca tcagcgagcg
      181 atatccgtgc ctcagcgaat tggcgcggga gcactgcata cccgagttcg acaatctgta
      241 cttggacatg aacggcattg tccacaactg ctcgcatccg gacgacaaca acatacactt
      301 tcacctggag gaggagcaga tattccagga aatcttcaac tacgtagaca agctgttcta
      361 cctgatcaag ccgcagcgcc tcttcttcct ttcagttgac ggggtggctc cacgggccaa
      421 gatgaaccag cagcgcagcc ggcgattccg gacggcgcgc gaggcggagc agcaggaggc
      481 caaggcggcc cagcgaggcg agttgcgaga gcacgagcgc ttcgacagca actgcattac
      541 gccgggcacc gaattcatgg tgcgtctgca ggagggactg cgcgcctttc tcaaaacgaa
      601 gatttcgacg gatccgctgt ggcagcgttg caccgtaatc ctaagtggcc aggaggcgcc
      661 cggcgagggc gagcacaaga tcatggacta cattcggtac atgaagacgc agccggacta
      721 tgatcccaat acacgacact gcctttacgg cctggacgcc gatctgatca ttctgggcct
      781 ctgcacccac gagctgcact tcgtggtgct gcgcgaggag gtcaagttcg ggcgcaacgt
      841 caagcgcacc tcggtggagg agacgcgctt cttcctgctg cacctgggtc tgctgcgcga
      901 gtacctcgag ctggagttcg acgccctgag aacagacgag cacaaattgg acattgcgca
      961 gcttatcgac gactgggtgc tgatgggctt tttggtcggc aacgacttca ttccccacct
     1021 gccgtgcctg cacatttcat caaatgcact gccgctgctc taccgcacct acatcggcat
     1081 ttacccgacc ctcggcggca acatcaacga gaacggcaag ctcaacctgc gccgcttgca
     1141 gattttcata tcggccctca ccgaggtgga gctggatcac ttcaaggagc acgccgacga
     1201 cttgaagtac atgaacaaca aatcagaggc gttcgacatg gacgtcggcg agatcaccga
     1261 gtctcaaaac ctcgactccg atctcggagc cctcatcaac aagagcatgc tgctctacga
     1321 tgacgacagc gaggaggact gctccgacga gaacgcggtt cttctgaagg agttccagaa
     1381 ctacaagcga aacttctacc gcaacaagtt caagcgtgat ccgaatgatg aactgatcga
     1441 ggaactgtgc caccactacg tcaatgcgct gcagtgggtg ctcgactact actaccgtgg
     1501 cgtgcagtcc tgggactggt actatccctt ccactacact ccattcatca gcgacctcaa
     1561 gaacatcgag caggtggaga tcgccttcca catgggcacg ccctttctgc cattccagca
     1621 gcttctggcg gtactgccgg cggccagtgc aaagcttctc cctgtcgcct accacgacct
     1681 catgctgctg ccgaccagcc cgctggccga gttctatcca ctggagttcg agagcgacct
     1741 caatggcaag aaacacgatt gggaggcggt tgtgctgata cccttcatcg atgagggtcg
     1801 gctgctggcc gctatgctgc catgcgaggc tcagctgtcg ctggaggagc gcgaacgcaa
     1861 ctaggcggcc gcataatgct taagtcgaac agaaagtaat cgtattgtac acggccgcat
     1921 aatcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tccccatctt
     1981 agtatattag ttaagtataa gaaggagata tacataatat gtcccctata ctaggttatt
     2041 ggaaaattaa gggccttgtg caacccactc gacttctttt ggaatatctt gaagaaaaat
     2101 atgaagagca tttgtatgag cgcgatgaag gtgataaatg gcgaaacaaa aagtttgaat
     2161 tgggtttgga gtttcccaat cttccttatt atattgatgg tgatgttaaa ttaacacagt
     2221 ctatggccat catacgttat atagctgaca agcacaacat gttgggtggt tgtccaaaag
     2281 agcgtgcaga gatttcaatg cttgaaggag cggttttgga tattagatac ggtgtttcga
     2341 gaattgcata tagtaaagac tttgaaactc tcaaagttga ttttcttagc aagctacctg
     2401 aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac atatttaaat ggtgatcatg
     2461 taacccatcc tgacttcatg ttgtatgacg ctcttgatgt tgttttatac atggacccaa
     2521 tgtgcctgga tgcgttccca aaattagttt gttttaaaaa acgtattgaa gctatcccac
     2581 aaattgataa gtacttgaaa tccagcaagt atatagcatg gcctttgcag ggctggcaag
     2641 ccacgtttgg tggtggcgac catcctccaa aatcggatct ggaagttctg ttccaggggc
     2701 cccatatggc agatctcaat tggatatcgg ccggccacgc gatcgctgac gtcggtaccc
     2761 tcgagtctgg taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgtcta
     2821 ctagcgcagc ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc
     2881 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc
     2941 acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg
     3001 ctatttaacg accctgccct gaaccgacga caagctgacg accgggtctc cgcaagtggc
     3061 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3121 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     3181 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     3241 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     3301 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     3361 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     3421 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     3481 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcggt cgctgttaaa
     3541 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     3601 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     3661 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     3721 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     3781 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     3841 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     3901 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     3961 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4021 catatttgaa tgtatttaga aaaataaaca aataggcatg cagcgctctt ccgcttcctc
     4081 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggtgtcag ctcactcaaa
     4141 agcggtaata cggttatcca cagaatcagg ggataaagcc ggaaagaaca tgtgagcaaa
     4201 aagcaaagca ccggaagaag ccaacgccgc aggcgttttt ccataggctc cgcccccctg
     4261 acgagcatca caaaaatcga cgctcaagcc agaggtggcg aaacccgaca ggactataaa
     4321 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4381 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4441 gctgttggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4501 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4561 taagacacga cttatcgcca ctggcagcag ccattggtaa ctgatttaga ggactttgtc
     4621 ttgaagttat gcacctgtta aggctaaact gaaagaacag attttggtga gtgcggtcct
     4681 ccaacccact taccttggtt caaagagttg gtagctcagc gaaccttgag aaaaccaccg
     4741 ttggtagcgg tggtttttct ttatttatga gatgatgaat caatcggtct atcaagtcaa
     4801 cgaacagcta ttccgttact ctagatttca gtgcaattta tctcttcaaa tgtagcacct
     4861 gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc
     4921 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     4981 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5041 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5101 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     5161 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     5221 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     5281 cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     5341 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     5401 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     5461 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     5521 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     5581 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     5641 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     5701 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     5761 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     5821 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     5881 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     5941 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     6001 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     6061 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     6121 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     6181 tcgacgctct cccttatgcg actcctgcat taggaaatta atacgactca ctata
//
