LOCUS       sequence-288416-        5047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsRck_296-472.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5047)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5047)
  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_296-472(MCSI)_B
FEATURES             Location/Qualifiers
     source          1..5047
                     /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        742..760
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    761..785
                     /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             831..1484
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1442..1464
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1491..1514
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1584..1628
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1666..1684)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1680..1727
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1957..2772)
                     /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(1957..2772)
                     /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     2680..2699
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2773..2864)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2880..3629)
                     /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(2890..2907)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3130..3149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3825..4907)
                     /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     4869..4888
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4908..4985)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5031..5047,1..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 cgaagggagt aacccagtac tacgcatatg taactgagcg
      181 ccaaaaagta cactgcctca acacactttt ctccaggctt cagataaacc agtcgatcat
      241 tttctgtaac tcctctcagc gagttgaatt gctagccaag aagatttctc aactgggtta
      301 ttcttgcttc tatattcatg ctaaaatgag gcaggaacat cgaaatcgtg tatttcatga
      361 tttccgaaat ggcttatgcc gcaatcttgt ttgcactgat ctgtttaccc gaggtattga
      421 tatacaagct gtgaatgtgg taataaactt tgatttccca aagctggcag agacctatct
      481 ccatcgtatt ggaagatcag gtcgctttgg tcatcttggc ttagccatca acttgatcac
      541 atatgatgat cgcttcaacc tgaaaagtat tgaggagcag ctgggaacag aaattaaacc
      601 tattccgagc aacattgata agagcctgta tgtggcagaa taccacagcg agcctgtaga
      661 agatgagaaa ccttaagcgg ccgcataatg cttaagtcga acagaaagta atcgtattgt
      721 acacggccgc ataatcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
      781 attccccatc ttagtatatt agttaagtat aagaaggaga tatacataat atgtccccta
      841 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
      901 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
      961 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
     1021 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
     1081 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
     1141 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
     1201 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
     1261 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
     1321 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
     1381 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
     1441 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc
     1501 tgttccaggg gccccatatg gcagatctca attggatatc ggccggccac gcgatcgctg
     1561 acgtcggtac cctcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca
     1621 tggactcgtc tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac
     1681 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg
     1741 catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat
     1801 agacataagc ggctatttaa cgaccctgcc ctgaaccgac gacaagctga cgaccgggtc
     1861 tccgcaagtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1921 atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa
     1981 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     2041 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     2101 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     2161 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     2221 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     2281 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     2341 gtcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     2401 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     2461 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     2521 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     2581 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     2641 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     2701 atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg
     2761 aatatggctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     2821 catgagcgga tacatatttg aatgtattta gaaaaataaa caaataggca tgcagcgctc
     2881 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggtgtc
     2941 agctcactca aaagcggtaa tacggttatc cacagaatca ggggataaag ccggaaagaa
     3001 catgtgagca aaaagcaaag caccggaaga agccaacgcc gcaggcgttt ttccataggc
     3061 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag ccagaggtgg cgaaacccga
     3121 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3181 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3241 ctcatagctc acgctgttgg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3301 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3361 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccattggt aactgattta
     3421 gaggactttg tcttgaagtt atgcacctgt taaggctaaa ctgaaagaac agattttggt
     3481 gagtgcggtc ctccaaccca cttaccttgg ttcaaagagt tggtagctca gcgaaccttg
     3541 agaaaaccac cgttggtagc ggtggttttt ctttatttat gagatgatga atcaatcggt
     3601 ctatcaagtc aacgaacagc tattccgtta ctctagattt cagtgcaatt tatctcttca
     3661 aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc
     3721 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
     3781 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     3841 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3901 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     3961 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     4021 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     4081 gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg
     4141 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     4201 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     4261 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     4321 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     4381 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     4441 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     4501 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     4561 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     4621 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     4681 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     4741 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     4801 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     4861 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     4921 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     4981 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact
     5041 cactata
//
