LOCUS       sequence-288558-        4942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsPatL1_399-770.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4942)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 4942)
  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_399-770_D
FEATURES             Location/Qualifiers
     source          1..4942
                     /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        1327..1345
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1346..1370
                     /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             1479..1523
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1561..1579)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1575..1622
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1852..2667)
                     /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(1852..2667)
                     /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     2575..2594
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2668..2759)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2775..3524)
                     /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(2785..2802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3025..3044)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3720..4802)
                     /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     4764..4783
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4803..4880)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4926..4942,1..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 tggatccata tgccaatctc atgttgcagc gggaaaagga
      181 ttgggtctct aaaatccaga tgatgcaact gcaaagcact gatccctacc tggatgattt
      241 ttattaccag aattactttg aaaaactgga gaaactgtca gctgctgaag aaatacaagg
      301 tgatggccct aagaaggagc gcaccaagct tatcacccct caggtggcca aactggagca
      361 cgcctataag ccagtgcaat ttgagggctc tttgggaaag cttaccgttt ctagtgtgaa
      421 taatccccga aaaatgattg atgctgttgt gacatctcgg agtgaggatg atgagacaaa
      481 agaaaaacaa gttcgagaca agaggagaaa aacccttgtt ataattgaga aaacctacag
      541 cttactcctt gatgtggagg actatgaaag acgttatctc ctaagtctgg aagaagagcg
      601 acctgcccta atggatgaca gaaagcacaa aatttgtagc atgtatgaca acttaagggg
      661 gaaattgcct ggacaagaga ggcctagtga tgaccacttt gtacagatca tgtgtatccg
      721 aaaagggaag agaatggttg cccgtattct tcctttcctc tccacagagc aagcagctga
      781 cattctcatg acaacagcca ggaacctccc tttccttatc aagaaggatg cacaagatga
      841 ggtgctgcca tgcttactga gtcccttctc tctccttctc tatcatcttc catcagtgag
      901 tatcaccagc cttttgcgac agctaatgaa cctacctcaa agtgcagcta caccagcact
      961 ctccaatcct cacctcactg ctgtgctcca gaacaagttt ggcctgtcac tgctcctcat
     1021 cctcctgagc cgtggtgaag acctacagag ttcagaccct gctacagaat caacacaaaa
     1081 taatcagtgg acggaggtga tgttcatggc aacacgagaa cttctgcgga ttccccaagc
     1141 agccctggcc aagccaatct ctatacctac aaacctagtg tccctctttt ctcgctatgt
     1201 tgaccggcag aaactgaact tgctggagac aaaactgcag ctagttcagg ggatacgata
     1261 agcggccgca taatgcttaa gtcgaacaga aagtaatcgt attgtacacg gccgcataat
     1321 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc ccatcttagt
     1381 atattagtta agtataagaa ggagatatac atatggcaga tctcaattgg atatcggccg
     1441 gccacgcgat cgctgacgtc ggtaccctcg agtctggtaa agaaaccgct gctgcgaaat
     1501 ttgaacgcca gcacatggac tcgtctacta gcgcagctta attaacctag gctgctgcca
     1561 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1621 tgctgaaacc tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac
     1681 cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgacaa
     1741 gctgacgacc gggtctccgc aagtggcact tttcggggaa atgtgcgcgg aacccctatt
     1801 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa
     1861 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     1921 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     1981 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     2041 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     2101 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     2161 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     2221 gagacgaaat acgcggtcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     2281 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     2341 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     2401 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     2461 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     2521 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     2581 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca
     2641 agacgtttcc cgttgaatat ggctcatact cttccttttt caatattatt gaagcattta
     2701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2761 aggcatgcag cgctcttccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact
     2821 gcggcgagcg gtgtcagctc actcaaaagc ggtaatacgg ttatccacag aatcagggga
     2881 taaagccgga aagaacatgt gagcaaaaag caaagcaccg gaagaagcca acgccgcagg
     2941 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagccaga
     3001 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3061 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3121 gaagcgtggc gctttctcat agctcacgct gttggtatct cagttcggtg taggtcgttc
     3181 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3241 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3301 ttggtaactg atttagagga ctttgtcttg aagttatgca cctgttaagg ctaaactgaa
     3361 agaacagatt ttggtgagtg cggtcctcca acccacttac cttggttcaa agagttggta
     3421 gctcagcgaa ccttgagaaa accaccgttg gtagcggtgg tttttcttta tttatgagat
     3481 gatgaatcaa tcggtctatc aagtcaacga acagctattc cgttactcta gatttcagtg
     3541 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc
     3601 tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     3661 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     3721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3781 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     3841 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     3901 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     3961 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     4021 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     4081 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     4141 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     4201 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     4261 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     4321 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     4381 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     4441 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     4501 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     4561 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     4621 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     4681 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     4741 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     4801 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     4861 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     4921 gaaattaata cgactcacta ta
//
