LOCUS       sequence-288413-        4855 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 4855)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 4855)
  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_1-111 (MCSII)_B
FEATURES             Location/Qualifiers
     source          1..4855
                     /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             1392..1436
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1474..1492)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1488..1535
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1765..2580)
                     /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(1765..2580)
                     /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     2488..2507
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2581..2672)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2688..3437)
                     /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(2698..2715)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2938..2957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3633..4715)
                     /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     4677..4696
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4716..4793)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4839..4855,1..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 tagggtaccc
     1381 tcgagtctgg taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgtcta
     1441 ctagcgcagc ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc
     1501 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc
     1561 acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg
     1621 ctatttaacg accctgccct gaaccgacga caagctgacg accgggtctc cgcaagtggc
     1681 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1741 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     1801 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     1861 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     1921 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     1981 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     2041 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     2101 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcggt cgctgttaaa
     2161 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     2221 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     2281 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     2341 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     2401 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     2461 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     2521 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     2581 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2641 catatttgaa tgtatttaga aaaataaaca aataggcatg cagcgctctt ccgcttcctc
     2701 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggtgtcag ctcactcaaa
     2761 agcggtaata cggttatcca cagaatcagg ggataaagcc ggaaagaaca tgtgagcaaa
     2821 aagcaaagca ccggaagaag ccaacgccgc aggcgttttt ccataggctc cgcccccctg
     2881 acgagcatca caaaaatcga cgctcaagcc agaggtggcg aaacccgaca ggactataaa
     2941 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3001 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3061 gctgttggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3121 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3181 taagacacga cttatcgcca ctggcagcag ccattggtaa ctgatttaga ggactttgtc
     3241 ttgaagttat gcacctgtta aggctaaact gaaagaacag attttggtga gtgcggtcct
     3301 ccaacccact taccttggtt caaagagttg gtagctcagc gaaccttgag aaaaccaccg
     3361 ttggtagcgg tggtttttct ttatttatga gatgatgaat caatcggtct atcaagtcaa
     3421 cgaacagcta ttccgttact ctagatttca gtgcaattta tctcttcaaa tgtagcacct
     3481 gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc
     3541 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     3601 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3661 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3721 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     3781 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     3841 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     3901 cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     3961 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     4021 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     4081 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     4141 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     4201 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     4261 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     4321 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     4381 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     4441 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     4501 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     4561 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     4621 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     4681 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     4741 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     4801 tcgacgctct cccttatgcg actcctgcat taggaaatta atacgactca ctata
//
