LOCUS       sequence-288559-        4153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsPatL1_408-516.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4153)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 4153)
  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_408-516_D
FEATURES             Location/Qualifiers
     source          1..4153
                     /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        538..556
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    557..581
                     /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             690..734
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(772..790)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      786..833
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1063..1878)
                     /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(1063..1878)
                     /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     1786..1805
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1879..1970)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1986..2735)
                     /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(1996..2013)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2236..2255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2931..4013)
                     /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     3975..3994
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4014..4091)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4137..4153,1..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 caagttcgat aggcggccgc
      481 ataatgctta agtcgaacag aaagtaatcg tattgtacac ggccgcataa tcgaaattaa
      541 tacgactcac tataggggaa ttgtgagcgg ataacaattc cccatcttag tatattagtt
      601 aagtataaga aggagatata catatggcag atctcaattg gatatcggcc ggccacgcga
      661 tcgctgacgt cggtaccctc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc
      721 agcacatgga ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc
      781 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac
      841 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
      901 agcaatagac ataagcggct atttaacgac cctgccctga accgacgaca agctgacgac
      961 cgggtctccg caagtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1021 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     1081 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     1141 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     1201 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     1261 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     1321 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     1381 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     1441 tacgcggtcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     1501 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     1561 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     1621 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     1681 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     1741 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     1801 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     1861 ccgttgaata tggctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1921 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggcatgca
     1981 gcgctcttcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     2041 ggtgtcagct cactcaaaag cggtaatacg gttatccaca gaatcagggg ataaagccgg
     2101 aaagaacatg tgagcaaaaa gcaaagcacc ggaagaagcc aacgccgcag gcgtttttcc
     2161 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagccag aggtggcgaa
     2221 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2281 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2341 cgctttctca tagctcacgc tgttggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2401 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2461 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc attggtaact
     2521 gatttagagg actttgtctt gaagttatgc acctgttaag gctaaactga aagaacagat
     2581 tttggtgagt gcggtcctcc aacccactta ccttggttca aagagttggt agctcagcga
     2641 accttgagaa aaccaccgtt ggtagcggtg gtttttcttt atttatgaga tgatgaatca
     2701 atcggtctat caagtcaacg aacagctatt ccgttactct agatttcagt gcaatttatc
     2761 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag
     2821 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     2881 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     2941 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3001 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     3061 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     3121 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     3181 tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg
     3241 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     3301 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     3361 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     3421 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     3481 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     3541 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     3601 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     3661 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     3721 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     3781 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     3841 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     3901 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     3961 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     4021 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     4081 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaaattaat
     4141 acgactcact ata
//
