LOCUS       sequence-288560-        4915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsPatL1_408-770.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4915)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 4915)
  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: pRSFDuetP-HsPatL1_408-770_D
FEATURES             Location/Qualifiers
     source          1..4915
                     /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        1300..1318
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1319..1343
                     /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             1452..1496
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1534..1552)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1548..1595
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1825..2640)
                     /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(1825..2640)
                     /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     2548..2567
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2641..2732)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2748..3497)
                     /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(2758..2775)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2998..3017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3693..4775)
                     /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     4737..4756
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4776..4853)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4899..4915,1..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 cgcgggaaaa ggattgggtc tctaaaatcc agatgatgca
      181 actgcaaagc actgatccct acctggatga tttttattac cagaattact ttgaaaaact
      241 ggagaaactg tcagctgctg aagaaataca aggtgatggc cctaagaagg agcgcaccaa
      301 gcttatcacc cctcaggtgg ccaaactgga gcacgcctat aagccagtgc aatttgaggg
      361 ctctttggga aagcttaccg tttctagtgt gaataatccc cgaaaaatga ttgatgctgt
      421 tgtgacatct cggagtgagg atgatgagac aaaagaaaaa caagttcgag acaagaggag
      481 aaaaaccctt gttataattg agaaaaccta cagcttactc cttgatgtgg aggactatga
      541 aagacgttat ctcctaagtc tggaagaaga gcgacctgcc ctaatggatg acagaaagca
      601 caaaatttgt agcatgtatg acaacttaag ggggaaattg cctggacaag agaggcctag
      661 tgatgaccac tttgtacaga tcatgtgtat ccgaaaaggg aagagaatgg ttgcccgtat
      721 tcttcctttc ctctccacag agcaagcagc tgacattctc atgacaacag ccaggaacct
      781 ccctttcctt atcaagaagg atgcacaaga tgaggtgctg ccatgcttac tgagtccctt
      841 ctctctcctt ctctatcatc ttccatcagt gagtatcacc agccttttgc gacagctaat
      901 gaacctacct caaagtgcag ctacaccagc actctccaat cctcacctca ctgctgtgct
      961 ccagaacaag tttggcctgt cactgctcct catcctcctg agccgtggtg aagacctaca
     1021 gagttcagac cctgctacag aatcaacaca aaataatcag tggacggagg tgatgttcat
     1081 ggcaacacga gaacttctgc ggattcccca agcagccctg gccaagccaa tctctatacc
     1141 tacaaaccta gtgtccctct tttctcgcta tgttgaccgg cagaaactga acttgctgga
     1201 gacaaaactg cagctagttc aggggatacg ataagcggcc gcataatgct taagtcgaac
     1261 agaaagtaat cgtattgtac acggccgcat aatcgaaatt aatacgactc actatagggg
     1321 aattgtgagc ggataacaat tccccatctt agtatattag ttaagtataa gaaggagata
     1381 tacatatggc agatctcaat tggatatcgg ccggccacgc gatcgctgac gtcggtaccc
     1441 tcgagtctgg taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgtcta
     1501 ctagcgcagc ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc
     1561 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc
     1621 acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg
     1681 ctatttaacg accctgccct gaaccgacga caagctgacg accgggtctc cgcaagtggc
     1741 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1801 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     1861 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     1921 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     1981 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     2041 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     2101 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     2161 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcggt cgctgttaaa
     2221 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     2281 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     2341 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     2401 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     2461 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     2521 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     2581 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     2641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2701 catatttgaa tgtatttaga aaaataaaca aataggcatg cagcgctctt ccgcttcctc
     2761 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggtgtcag ctcactcaaa
     2821 agcggtaata cggttatcca cagaatcagg ggataaagcc ggaaagaaca tgtgagcaaa
     2881 aagcaaagca ccggaagaag ccaacgccgc aggcgttttt ccataggctc cgcccccctg
     2941 acgagcatca caaaaatcga cgctcaagcc agaggtggcg aaacccgaca ggactataaa
     3001 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3061 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3121 gctgttggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3181 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3241 taagacacga cttatcgcca ctggcagcag ccattggtaa ctgatttaga ggactttgtc
     3301 ttgaagttat gcacctgtta aggctaaact gaaagaacag attttggtga gtgcggtcct
     3361 ccaacccact taccttggtt caaagagttg gtagctcagc gaaccttgag aaaaccaccg
     3421 ttggtagcgg tggtttttct ttatttatga gatgatgaat caatcggtct atcaagtcaa
     3481 cgaacagcta ttccgttact ctagatttca gtgcaattta tctcttcaaa tgtagcacct
     3541 gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc
     3601 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     3661 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3721 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3781 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     3841 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     3901 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     3961 cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     4021 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     4081 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     4141 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     4201 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     4261 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     4321 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     4381 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     4441 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     4501 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     4561 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     4621 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     4681 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     4741 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     4801 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     4861 tcgacgctct cccttatgcg actcctgcat taggaaatta atacgactca ctata
//
