LOCUS       sequence-383963-        6082 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 6082)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6082)
  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..6082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     248..270
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             297..320
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             540..584
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(622..640)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      636..683
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(913..1728)
                     /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(913..1728)
                     /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     1636..1655
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1729..1820)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1836..2585)
                     /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(1846..1863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2086..2105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2781..3863)
                     /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     3825..3844
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3864..3941)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        3987..4005
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4006..4030
                     /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             4086..4103
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4113..4136
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        5630..5648
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5649..5673
                     /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             join(5719..6082,1..290)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
ORIGIN
        1 ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat
       61 atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg
      121 ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac
      181 gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc
      241 ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tcggatctgg
      301 aagttctgtt ccaggggccc catatggcag atctcaattg gatgttccgc tacgagtctt
      361 tggaggattg tcctctggat gaagatgaag atgcatttca gggactggga gaagaagatg
      421 aagagattga tcaattcaat gatgatacat ttgggtcagg tgcagttgat gatgattggc
      481 aggaagcaca tgagcgcctg gctgaattgg aagaaaagta gggtaccctc gagtctggta
      541 aagaaaccgc tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt
      601 aattaaccta ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
      661 aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac
      721 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
      781 cctgccctga accgacgaca agctgacgac cgggtctccg caagtggcac ttttcgggga
      841 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      901 atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
      961 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     1021 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     1081 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     1141 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     1201 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     1261 cattcgtgat tgcgcctgag cgagacgaaa tacgcggtcg ctgttaaaag gacaattaca
     1321 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     1381 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     1441 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     1501 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     1561 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     1621 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     1681 atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcatac tcttcctttt
     1741 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1801 tatttagaaa aataaacaaa taggcatgca gcgctcttcc gcttcctcgc tcactgactc
     1861 gctacgctcg gtcgttcgac tgcggcgagc ggtgtcagct cactcaaaag cggtaatacg
     1921 gttatccaca gaatcagggg ataaagccgg aaagaacatg tgagcaaaaa gcaaagcacc
     1981 ggaagaagcc aacgccgcag gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2041 aaaatcgacg ctcaagccag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2101 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2161 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgttggtatc
     2221 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2281 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2341 tatcgccact ggcagcagcc attggtaact gatttagagg actttgtctt gaagttatgc
     2401 acctgttaag gctaaactga aagaacagat tttggtgagt gcggtcctcc aacccactta
     2461 ccttggttca aagagttggt agctcagcga accttgagaa aaccaccgtt ggtagcggtg
     2521 gtttttcttt atttatgaga tgatgaatca atcggtctat caagtcaacg aacagctatt
     2581 ccgttactct agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc
     2641 atacgatata agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga
     2701 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     2761 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2821 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     2881 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     2941 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     3001 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     3061 atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     3121 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     3181 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     3241 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     3301 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     3361 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     3421 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     3481 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     3541 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     3601 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     3661 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     3721 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     3781 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     3841 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     3901 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     3961 cttatgcgac tcctgcatta ggaaattaat acgactcact ataggggaat tgtgagcgga
     4021 taacaattcc cctgtagaaa taattttgtt taactttaat aaggagatat accatgggca
     4081 gcagccatca ccatcatcac cacagcgcgg gtctggaagt tctgttccag gggccccagg
     4141 atccgatggg tctgtccagt caaaatggtc agctgagagg ccctgtgaaa cccactggtg
     4201 gccctggagg agggggcaca cagacacagc aacagatgaa ccagctgaaa aacaccaaca
     4261 caatcaataa tggcactcag cagcaagcac agagtatgac caccactatt aaacctggtg
     4321 atgactggaa aaagacttta aaactccctc caaaggatct aagaatcaaa acttcggatg
     4381 tgacctccac aaaaggaaat gagtttgaag attactgttt gaaacgggag ttactgatgg
     4441 gaatttttga aatgggctgg gaaaagccat ctcctattca ggaggagagc attcccattg
     4501 ctttatctgg tagggatatc ttagctagag caaaaaatgg aacaggcaag agcggtgcct
     4561 acctcattcc cttacttgaa cggctagacc tgaagaagga caatatacaa gcaatggtga
     4621 ttgttcccac tagagaactt gctctacagg tcagtcaaat ttgcatccag gtcagcaaac
     4681 acatgggagg ggccaaagtg atggcaacca caggaggaac caatttacga gatgacataa
     4741 tgaggcttga tgatacagtg cacgtggtga ttgctacccc tgggagaatc ctggatctta
     4801 ttaagaaagg agtagcaaag gttgatcatg tccagatgat agtattggat gaggcagata
     4861 agttgctgtc acaggatttt gtgcagataa tggaggatat tattctcacg ctacctaaaa
     4921 acaggcagat tttactatat tccgctactt tccctcttag tgtacagaag ttcatgaatt
     4981 cccatttgca gaaaccctat gagattaacc tgatggagga actaactctg aagggagtaa
     5041 cccagtacta cgcatatgta actgagcgcc aaaaagtaca ctgcctcaac acacttttct
     5101 ccaggcttca gataaaccag tcgatcattt tctgtaactc ctctcagcga gttgaattgc
     5161 tagccaagaa gatttctcaa ctgggttatt cttgcttcta tattcatgct aaaatgaggc
     5221 aggaacatcg aaatcgtgta tttcatgatt tccgaaatgg cttatgccgc aatcttgttt
     5281 gcactgatct gtttacccga ggtattgata tacaagctgt gaatgtggta ataaactttg
     5341 atttcccaaa gctggcagag acctatctcc atcgtattgg aagatcaggt cgctttggtc
     5401 atcttggctt agccatcaac ttgatcacat atgatgatcg cttcaacctg aaaagtattg
     5461 aggagcagct gggaacagaa attaaaccta ttccgagcaa cattgataag agcctgtatg
     5521 tggcagaata ccacagcgag cctgtagaag atgagaaacc ttaagcggcc gcataatgct
     5581 taagtcgaac agaaagtaat cgtattgtac acggccgcat aatcgaaatt aatacgactc
     5641 actatagggg aattgtgagc ggataacaat tccccatctt agtatattag ttaagtataa
     5701 gaaggagata tacataatat gtcccctata ctaggttatt ggaaaattaa gggccttgtg
     5761 caacccactc gacttctttt ggaatatctt gaagaaaaat atgaagagca tttgtatgag
     5821 cgcgatgaag gtgataaatg gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat
     5881 cttccttatt atattgatgg tgatgttaaa ttaacacagt ctatggccat catacgttat
     5941 atagctgaca agcacaacat gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg
     6001 cttgaaggag cggttttgga tattagatac ggtgtttcga gaattgcata tagtaaagac
     6061 tttgaaactc tcaaagttga tt
//
