LOCUS       sequence-288419-        6236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsPatL1_1-111.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6236)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6236)
  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_1-111_B
FEATURES             Location/Qualifiers
     source          1..6236
                     /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             2773..2817
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2855..2873)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2869..2916
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3146..3961)
                     /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(3146..3961)
                     /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     3869..3888
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3962..4053)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4069..4818)
                     /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(4079..4096)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4319..4338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5014..6096)
                     /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     6058..6077
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6097..6174)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6220..6236,1..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 aggcggtacc
     2761 ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac gccagcacat ggactcgtct
     2821 actagcgcag cttaattaac ctaggctgct gccaccgctg agcaataact agcataaccc
     2881 cttggggcct ctaaacgggt cttgaggggt tttttgctga aacctcaggc atttgagaag
     2941 cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg
     3001 gctatttaac gaccctgccc tgaaccgacg acaagctgac gaccgggtct ccgcaagtgg
     3061 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3121 tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca
     3181 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     3241 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     3301 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     3361 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     3421 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     3481 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcgg tcgctgttaa
     3541 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     3601 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     3661 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     3721 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     3781 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     3841 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     3901 catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca
     3961 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4021 acatatttga atgtatttag aaaaataaac aaataggcat gcagcgctct tccgcttcct
     4081 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggtgtca gctcactcaa
     4141 aagcggtaat acggttatcc acagaatcag gggataaagc cggaaagaac atgtgagcaa
     4201 aaagcaaagc accggaagaa gccaacgccg caggcgtttt tccataggct ccgcccccct
     4261 gacgagcatc acaaaaatcg acgctcaagc cagaggtggc gaaacccgac aggactataa
     4321 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4381 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4441 cgctgttggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4501 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4561 gtaagacacg acttatcgcc actggcagca gccattggta actgatttag aggactttgt
     4621 cttgaagtta tgcacctgtt aaggctaaac tgaaagaaca gattttggtg agtgcggtcc
     4681 tccaacccac ttaccttggt tcaaagagtt ggtagctcag cgaaccttga gaaaaccacc
     4741 gttggtagcg gtggtttttc tttatttatg agatgatgaa tcaatcggtc tatcaagtca
     4801 acgaacagct attccgttac tctagatttc agtgcaattt atctcttcaa atgtagcacc
     4861 tgaagtcagc cccatacgat ataagttgta attctcatgt tagtcatgcc ccgcgcccac
     4921 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     4981 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5041 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5101 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     5161 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     5221 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     5281 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     5341 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     5401 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     5461 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     5521 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     5581 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     5641 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     5701 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     5761 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     5821 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     5881 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     5941 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     6001 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     6061 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     6121 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     6181 ctcgacgctc tcccttatgc gactcctgca ttaggaaatt aatacgactc actata
//
