LOCUS       sequence-288415-        5716 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 5716)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5716)
  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-HsPatL1_1-398 (MCSII)_B
FEATURES             Location/Qualifiers
     source          1..5716
                     /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        244..262
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    263..287
                     /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             333..986
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     944..966
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             993..1016
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2253..2297
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2335..2353)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2349..2396
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2626..3441)
                     /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(2626..3441)
                     /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     3349..3368
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3442..3533)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3549..4298)
                     /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(3559..3576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3799..3818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4494..5576)
                     /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     5538..5557
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5577..5654)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5700..5716,1..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 cgaattcgag ctcggcgcgc ctgcaggtcg acaagcttgc
      181 ggccgcataa tgcttaagtc gaacagaaag taatcgtatt gtacacggcc gcataatcga
      241 aattaatacg actcactata ggggaattgt gagcggataa caattcccca tcttagtata
      301 ttagttaagt ataagaagga gatatacata atatgtcccc tatactaggt tattggaaaa
      361 ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag
      421 agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt
      481 tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg
      541 ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg
      601 cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg
      661 catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc
      721 tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc
      781 atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc
      841 tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg
      901 ataagtactt gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt
      961 ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt tctgttccag gggccccata
     1021 tggcagatct caattggatg ttccgctacg agtctttgga ggattgtcct ctggatgaag
     1081 atgaagatgc atttcaggga ctgggagaag aagatgaaga gattgatcaa ttcaatgatg
     1141 atacatttgg gtcaggtgca gttgatgatg attggcagga agcacatgag cgcctggctg
     1201 aattggaaga aaagctacca gtggcagtta atgaacaaac aggcaatgga gagagagatg
     1261 aaatggactt gttgggtgac catgaggaga atctggcaga aaggctcagt aagatggtga
     1321 ttgaaaatga actagaagat ccagctatta tgagggcagt gcagaccagg ccagttttac
     1381 aaccccaacc aggaagtctg aattccagta tctgggatgg atctgaagtt ctgaggcgaa
     1441 tccgaggacc actgcttgct caggaaatgc ctacagtgtc tgtattagaa tatgctttgc
     1501 ctcagaggcc cccccagggt ccagaagatg atcgggacct ttctgaacga gcattaccaa
     1561 ggcggtcaac ttcacctatc attggcagtc ctcctgttag agctgtcccc ataggcaccc
     1621 cacctaaaca gatggctgta cccagcttca cccaacagat tctgtgtccg aagcctgtcc
     1681 atgttcggcc cccaatgcca cctcgttatc ctgctcccta tggtgagagg atgtctccaa
     1741 accagctctg cagtgtcccg aactcttccc tcctgggtca cccttttcct cctagtgttc
     1801 ctcctgttct cagccccctc cagagagcac agcttcttgg aggagcacag ctacagcctg
     1861 gacggatgtc tcccagccag tttgcacggg tccctggatt tgttggtagt ccacttgctg
     1921 ccatgaatcc caagttgcta caagggcgag ttgggcagat gcttccccca gcaccaggct
     1981 tccgtgcctt ctttagtgct ccaccctccg ctacaccacc tccacagcag caccctcctg
     2041 gcccaggacc ccacctgcaa aacctaagat ctcaggcccc aatgtttaga ccggacacaa
     2101 ctcacctcca tccacagcac cgtcgactct tgcatcagag acagcaacag aatagaagtc
     2161 agcatcggaa tctcaatggt gcgggagata gaggaagtca ccggagcagt catcaagatc
     2221 atctccgaaa gtagggtacc ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac
     2281 gccagcacat ggactcgtct actagcgcag cttaattaac ctaggctgct gccaccgctg
     2341 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     2401 aacctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa
     2461 accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg acaagctgac
     2521 gaccgggtct ccgcaagtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2581 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
     2641 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     2701 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     2761 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     2821 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     2881 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     2941 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     3001 aaatacgcgg tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     3061 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     3121 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     3181 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     3241 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     3301 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     3361 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
     3421 ttcccgttga atatggctca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3481 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggcat
     3541 gcagcgctct tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg
     3601 agcggtgtca gctcactcaa aagcggtaat acggttatcc acagaatcag gggataaagc
     3661 cggaaagaac atgtgagcaa aaagcaaagc accggaagaa gccaacgccg caggcgtttt
     3721 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagc cagaggtggc
     3781 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3841 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3901 tggcgctttc tcatagctca cgctgttggt atctcagttc ggtgtaggtc gttcgctcca
     3961 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4021 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccattggta
     4081 actgatttag aggactttgt cttgaagtta tgcacctgtt aaggctaaac tgaaagaaca
     4141 gattttggtg agtgcggtcc tccaacccac ttaccttggt tcaaagagtt ggtagctcag
     4201 cgaaccttga gaaaaccacc gttggtagcg gtggtttttc tttatttatg agatgatgaa
     4261 tcaatcggtc tatcaagtca acgaacagct attccgttac tctagatttc agtgcaattt
     4321 atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt
     4381 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     4441 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     4501 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4561 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     4621 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     4681 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     4741 ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac
     4801 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     4861 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     4921 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     4981 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     5041 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     5101 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     5161 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     5221 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     5281 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     5341 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     5401 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     5461 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     5521 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     5581 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     5641 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaaatt
     5701 aatacgactc actata
//
