LOCUS       sequence-288418-        6082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsPatL1_1-59.
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-HsPatL1_1-59_B
FEATURES             Location/Qualifiers
     source          1..6082
                     /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             2619..2663
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2701..2719)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2715..2762
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2992..3807)
                     /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(2992..3807)
                     /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     3715..3734
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3808..3899)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3915..4664)
                     /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(3925..3942)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4165..4184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4860..5942)
                     /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     5904..5923
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5943..6020)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6066..6082,1..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 agaaaagtag ggtaccctcg agtctggtaa agaaaccgct gctgcgaaat
     2641 ttgaacgcca gcacatggac tcgtctacta gcgcagctta attaacctag gctgctgcca
     2701 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2761 tgctgaaacc tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac
     2821 cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgacaa
     2881 gctgacgacc gggtctccgc aagtggcact tttcggggaa atgtgcgcgg aacccctatt
     2941 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa
     3001 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     3061 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     3121 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     3181 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     3241 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     3301 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     3361 gagacgaaat acgcggtcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     3421 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     3481 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     3541 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     3601 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     3661 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     3721 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca
     3781 agacgtttcc cgttgaatat ggctcatact cttccttttt caatattatt gaagcattta
     3841 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3901 aggcatgcag cgctcttccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact
     3961 gcggcgagcg gtgtcagctc actcaaaagc ggtaatacgg ttatccacag aatcagggga
     4021 taaagccgga aagaacatgt gagcaaaaag caaagcaccg gaagaagcca acgccgcagg
     4081 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagccaga
     4141 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4201 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4261 gaagcgtggc gctttctcat agctcacgct gttggtatct cagttcggtg taggtcgttc
     4321 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4381 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4441 ttggtaactg atttagagga ctttgtcttg aagttatgca cctgttaagg ctaaactgaa
     4501 agaacagatt ttggtgagtg cggtcctcca acccacttac cttggttcaa agagttggta
     4561 gctcagcgaa ccttgagaaa accaccgttg gtagcggtgg tttttcttta tttatgagat
     4621 gatgaatcaa tcggtctatc aagtcaacga acagctattc cgttactcta gatttcagtg
     4681 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc
     4741 tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     4801 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     4861 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4921 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     4981 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     5041 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     5101 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     5161 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     5221 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     5281 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     5341 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     5401 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     5461 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     5521 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     5581 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     5641 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     5701 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     5761 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     5821 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     5881 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     5941 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     6001 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     6061 gaaattaata cgactcacta ta
//
