LOCUS       sequence-288414-        5041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsPatL1_1-173.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5041)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5041)
  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_1-173 (MCSII)_B
FEATURES             Location/Qualifiers
     source          1..5041
                     /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             1578..1622
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1660..1678)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1674..1721
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1951..2766)
                     /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(1951..2766)
                     /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     2674..2693
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2767..2858)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2874..3623)
                     /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(2884..2901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3124..3143)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3819..4901)
                     /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     4863..4882
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4902..4979)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5025..5041,1..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 gcattatagg
     1561 gtaccctcga gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag cacatggact
     1621 cgtctactag cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa taactagcat
     1681 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct caggcatttg
     1741 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
     1801 aagcggctat ttaacgaccc tgccctgaac cgacgacaag ctgacgaccg ggtctccgca
     1861 agtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     1921 tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa
     1981 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     2041 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     2101 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     2161 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     2221 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     2281 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcggtcgct
     2341 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     2401 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     2461 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     2521 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     2581 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     2641 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     2701 atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg
     2761 gctcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2821 cggatacata tttgaatgta tttagaaaaa taaacaaata ggcatgcagc gctcttccgc
     2881 ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg tgtcagctca
     2941 ctcaaaagcg gtaatacggt tatccacaga atcaggggat aaagccggaa agaacatgtg
     3001 agcaaaaagc aaagcaccgg aagaagccaa cgccgcaggc gtttttccat aggctccgcc
     3061 cccctgacga gcatcacaaa aatcgacgct caagccagag gtggcgaaac ccgacaggac
     3121 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3181 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3241 gctcacgctg ttggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3301 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3361 acccggtaag acacgactta tcgccactgg cagcagccat tggtaactga tttagaggac
     3421 tttgtcttga agttatgcac ctgttaaggc taaactgaaa gaacagattt tggtgagtgc
     3481 ggtcctccaa cccacttacc ttggttcaaa gagttggtag ctcagcgaac cttgagaaaa
     3541 ccaccgttgg tagcggtggt ttttctttat ttatgagatg atgaatcaat cggtctatca
     3601 agtcaacgaa cagctattcc gttactctag atttcagtgc aatttatctc ttcaaatgta
     3661 gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc atgccccgcg
     3721 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     3781 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3841 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3901 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     3961 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     4021 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     4081 tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc
     4141 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     4201 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     4261 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     4321 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     4381 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     4441 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     4501 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     4561 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     4621 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     4681 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     4741 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     4801 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     4861 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     4921 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     4981 gggatctcga cgctctccct tatgcgactc ctgcattagg aaattaatac gactcactat
     5041 a
//
