LOCUS       sequence-288417-        5932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsRck.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5932)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5932)
  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-HsRCK(MCSI)_B
FEATURES             Location/Qualifiers
     source          1..5932
                     /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        1627..1645
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1646..1670
                     /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             1716..2369
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2327..2349
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2376..2399
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2469..2513
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2551..2569)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2565..2612
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2842..3657)
                     /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(2842..3657)
                     /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     3565..3584
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3658..3749)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3765..4514)
                     /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(3775..3792)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4015..4034)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4710..5792)
                     /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     5754..5773
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5793..5870)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5916..5932,1..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 cgatgggtct gtccagtcaa aatggtcagc tgagaggccc
      181 tgtgaaaccc actggtggcc ctggaggagg gggcacacag acacagcaac agatgaacca
      241 gctgaaaaac accaacacaa tcaataatgg cactcagcag caagcacaga gtatgaccac
      301 cactattaaa cctggtgatg actggaaaaa gactttaaaa ctccctccaa aggatctaag
      361 aatcaaaact tcggatgtga cctccacaaa aggaaatgag tttgaagatt actgtttgaa
      421 acgggagtta ctgatgggaa tttttgaaat gggctgggaa aagccatctc ctattcagga
      481 ggagagcatt cccattgctt tatctggtag ggatatctta gctagagcaa aaaatggaac
      541 aggcaagagc ggtgcctacc tcattccctt acttgaacgg ctagacctga agaaggacaa
      601 tatacaagca atggtgattg ttcccactag agaacttgct ctacaggtca gtcaaatttg
      661 catccaggtc agcaaacaca tgggaggggc caaagtgatg gcaaccacag gaggaaccaa
      721 tttacgagat gacataatga ggcttgatga tacagtgcac gtggtgattg ctacccctgg
      781 gagaatcctg gatcttatta agaaaggagt agcaaaggtt gatcatgtcc agatgatagt
      841 attggatgag gcagataagt tgctgtcaca ggattttgtg cagataatgg aggatattat
      901 tctcacgcta cctaaaaaca ggcagatttt actatattcc gctactttcc ctcttagtgt
      961 acagaagttc atgaattccc atttgcagaa accctatgag attaacctga tggaggaact
     1021 aactctgaag ggagtaaccc agtactacgc atatgtaact gagcgccaaa aagtacactg
     1081 cctcaacaca cttttctcca ggcttcagat aaaccagtcg atcattttct gtaactcctc
     1141 tcagcgagtt gaattgctag ccaagaagat ttctcaactg ggttattctt gcttctatat
     1201 tcatgctaaa atgaggcagg aacatcgaaa tcgtgtattt catgatttcc gaaatggctt
     1261 atgccgcaat cttgtttgca ctgatctgtt tacccgaggt attgatatac aagctgtgaa
     1321 tgtggtaata aactttgatt tcccaaagct ggcagagacc tatctccatc gtattggaag
     1381 atcaggtcgc tttggtcatc ttggcttagc catcaacttg atcacatatg atgatcgctt
     1441 caacctgaaa agtattgagg agcagctggg aacagaaatt aaacctattc cgagcaacat
     1501 tgataagagc ctgtatgtgg cagaatacca cagcgagcct gtagaagatg agaaacctta
     1561 agcggccgca taatgcttaa gtcgaacaga aagtaatcgt attgtacacg gccgcataat
     1621 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc ccatcttagt
     1681 atattagtta agtataagaa ggagatatac ataatatgtc ccctatacta ggttattgga
     1741 aaattaaggg ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg
     1801 aagagcattt gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg
     1861 gtttggagtt tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta
     1921 tggccatcat acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc
     1981 gtgcagagat ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa
     2041 ttgcatatag taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa
     2101 tgctgaaaat gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa
     2161 cccatcctga cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt
     2221 gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa
     2281 ttgataagta cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca
     2341 cgtttggtgg tggcgaccat cctccaaaat cggatctgga agttctgttc caggggcccc
     2401 atatggcaga tctcaattgg atatcggccg gccacgcgat cgctgacgtc ggtaccctcg
     2461 agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta
     2521 gcgcagctta attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg
     2581 gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca
     2641 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
     2701 tttaacgacc ctgccctgaa ccgacgacaa gctgacgacc gggtctccgc aagtggcact
     2761 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2821 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     2881 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     2941 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     3001 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     3061 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     3121 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     3181 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcggtcgc tgttaaaagg
     3241 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     3301 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     3361 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     3421 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     3481 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     3541 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     3601 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcatact
     3661 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3721 atttgaatgt atttagaaaa ataaacaaat aggcatgcag cgctcttccg cttcctcgct
     3781 cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gtgtcagctc actcaaaagc
     3841 ggtaatacgg ttatccacag aatcagggga taaagccgga aagaacatgt gagcaaaaag
     3901 caaagcaccg gaagaagcca acgccgcagg cgtttttcca taggctccgc ccccctgacg
     3961 agcatcacaa aaatcgacgc tcaagccaga ggtggcgaaa cccgacagga ctataaagat
     4021 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4081 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4141 gttggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4201 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4261 gacacgactt atcgccactg gcagcagcca ttggtaactg atttagagga ctttgtcttg
     4321 aagttatgca cctgttaagg ctaaactgaa agaacagatt ttggtgagtg cggtcctcca
     4381 acccacttac cttggttcaa agagttggta gctcagcgaa ccttgagaaa accaccgttg
     4441 gtagcggtgg tttttcttta tttatgagat gatgaatcaa tcggtctatc aagtcaacga
     4501 acagctattc cgttactcta gatttcagtg caatttatct cttcaaatgt agcacctgaa
     4561 gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga
     4621 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     4681 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4741 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4801 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     4861 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     4921 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     4981 actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     5041 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     5101 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     5161 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     5221 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     5281 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     5341 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     5401 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     5461 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     5521 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     5581 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     5641 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     5701 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     5761 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     5821 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     5881 acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta ta
//
