LOCUS       sequence-288420-        6424 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial Expression of HsPatL1_1-173.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6424)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6424)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_1-173_B
FEATURES             Location/Qualifiers
     source          1..6424
                     /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             2961..3005
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3043..3061)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3057..3104
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3334..4149)
                     /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(3334..4149)
                     /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     4057..4076
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4150..4241)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4257..5006)
                     /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(4267..4284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4507..4526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5202..6284)
                     /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     6246..6265
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6285..6362)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6408..6424,1..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 aggccagttt
     2761 tacaacccca accaggaagt ctgaattcca gtatctggga tggatctgaa gttctgaggc
     2821 gaatccgagg accactgctt gctcaggaaa tgcctacagt gtctgtatta gaatatgctt
     2881 tgcctcagag gcccccccag ggtccagaag atgatcggga cctttctgaa cgagcattat
     2941 agggtaccct cgagtctggt aaagaaaccg ctgctgcgaa atttgaacgc cagcacatgg
     3001 actcgtctac tagcgcagct taattaacct aggctgctgc caccgctgag caataactag
     3061 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa cctcaggcat
     3121 ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga
     3181 cataagcggc tatttaacga ccctgccctg aaccgacgac aagctgacga ccgggtctcc
     3241 gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3301 cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg
     3361 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     3421 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     3481 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     3541 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     3601 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     3661 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcggtc
     3721 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     3781 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     3841 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     3901 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     3961 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     4021 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     4081 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     4141 atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4201 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggcatgc agcgctcttc
     4261 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggtgtcagc
     4321 tcactcaaaa gcggtaatac ggttatccac agaatcaggg gataaagccg gaaagaacat
     4381 gtgagcaaaa agcaaagcac cggaagaagc caacgccgca ggcgtttttc cataggctcc
     4441 gcccccctga cgagcatcac aaaaatcgac gctcaagcca gaggtggcga aacccgacag
     4501 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4561 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4621 atagctcacg ctgttggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4681 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4741 ccaacccggt aagacacgac ttatcgccac tggcagcagc cattggtaac tgatttagag
     4801 gactttgtct tgaagttatg cacctgttaa ggctaaactg aaagaacaga ttttggtgag
     4861 tgcggtcctc caacccactt accttggttc aaagagttgg tagctcagcg aaccttgaga
     4921 aaaccaccgt tggtagcggt ggtttttctt tatttatgag atgatgaatc aatcggtcta
     4981 tcaagtcaac gaacagctat tccgttactc tagatttcag tgcaatttat ctcttcaaat
     5041 gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgtta gtcatgcccc
     5101 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     5161 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     5221 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5281 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     5341 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     5401 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     5461 tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg
     5521 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     5581 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     5641 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     5701 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     5761 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     5821 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     5881 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     5941 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     6001 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     6061 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     6121 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     6181 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     6241 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     6301 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     6361 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaaattaa tacgactcac
     6421 tata
//
