LOCUS       sequence-288421-        7099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsPatL1_1-398.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7099)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7099)
  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-HsPatL1_1-398_B
FEATURES             Location/Qualifiers
     source          1..7099
                     /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             3636..3680
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3718..3736)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3732..3779
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4009..4824)
                     /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(4009..4824)
                     /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     4732..4751
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4825..4916)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4932..5681)
                     /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(4942..4959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5182..5201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5877..6959)
                     /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     6921..6940
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6960..7037)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(7083..7099,1..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 atgttccgct acgagtcttt ggaggattgt cctctggatg
     2461 aagatgaaga tgcatttcag ggactgggag aagaagatga agagattgat caattcaatg
     2521 atgatacatt tgggtcaggt gcagttgatg atgattggca ggaagcacat gagcgcctgg
     2581 ctgaattgga agaaaagcta ccagtggcag ttaatgaaca aacaggcaat ggagagagag
     2641 atgaaatgga cttgttgggt gaccatgagg agaatctggc agaaaggctc agtaagatgg
     2701 tgattgaaaa tgaactagaa gatccagcta ttatgagggc agtgcagacc aggccagttt
     2761 tacaacccca accaggaagt ctgaattcca gtatctggga tggatctgaa gttctgaggc
     2821 gaatccgagg accactgctt gctcaggaaa tgcctacagt gtctgtatta gaatatgctt
     2881 tgcctcagag gcccccccag ggtccagaag atgatcggga cctttctgaa cgagcattac
     2941 caaggcggtc aacttcacct atcattggca gtcctcctgt tagagctgtc cccataggca
     3001 ccccacctaa acagatggct gtacccagct tcacccaaca gattctgtgt ccgaagcctg
     3061 tccatgttcg gcccccaatg ccacctcgtt atcctgctcc ctatggtgag aggatgtctc
     3121 caaaccagct ctgcagtgtc ccgaactctt ccctcctggg tcaccctttt cctcctagtg
     3181 ttcctcctgt tctcagcccc ctccagagag cacagcttct tggaggagca cagctacagc
     3241 ctggacggat gtctcccagc cagtttgcac gggtccctgg atttgttggt agtccacttg
     3301 ctgccatgaa tcccaagttg ctacaagggc gagttgggca gatgcttccc ccagcaccag
     3361 gcttccgtgc cttctttagt gctccaccct ccgctacacc acctccacag cagcaccctc
     3421 ctggcccagg accccacctg caaaacctaa gatctcaggc cccaatgttt agaccggaca
     3481 caactcacct ccatccacag caccgtcgac tcttgcatca gagacagcaa cagaatagaa
     3541 gtcagcatcg gaatctcaat ggtgcgggag atagaggaag tcaccggagc agtcatcaag
     3601 atcatctccg aaagtagggt accctcgagt ctggtaaaga aaccgctgct gcgaaatttg
     3661 aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct gctgccaccg
     3721 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     3781 tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg
     3841 taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg acgacaagct
     3901 gacgaccggg tctccgcaag tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3961 ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc
     4021 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     4081 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     4141 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     4201 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     4261 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     4321 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     4381 acgaaatacg cggtcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     4441 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     4501 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     4561 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     4621 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     4681 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     4741 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga
     4801 cgtttcccgt tgaatatggc tcatactctt cctttttcaa tattattgaa gcatttatca
     4861 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4921 catgcagcgc tcttccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
     4981 gcgagcggtg tcagctcact caaaagcggt aatacggtta tccacagaat caggggataa
     5041 agccggaaag aacatgtgag caaaaagcaa agcaccggaa gaagccaacg ccgcaggcgt
     5101 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agccagaggt
     5161 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5221 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5281 gcgtggcgct ttctcatagc tcacgctgtt ggtatctcag ttcggtgtag gtcgttcgct
     5341 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5401 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccattg
     5461 gtaactgatt tagaggactt tgtcttgaag ttatgcacct gttaaggcta aactgaaaga
     5521 acagattttg gtgagtgcgg tcctccaacc cacttacctt ggttcaaaga gttggtagct
     5581 cagcgaacct tgagaaaacc accgttggta gcggtggttt ttctttattt atgagatgat
     5641 gaatcaatcg gtctatcaag tcaacgaaca gctattccgt tactctagat ttcagtgcaa
     5701 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca
     5761 tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     5821 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     5881 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5941 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     6001 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     6061 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     6121 gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg
     6181 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     6241 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     6301 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     6361 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     6421 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     6481 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     6541 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     6601 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     6661 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     6721 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     6781 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     6841 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     6901 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     6961 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     7021 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     7081 attaatacga ctcactata
//
