LOCUS       sequence-288553-        6580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsEDC3_192-508del231-254-HsRck_70-472.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6580)
  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-HsEDC3_192-508del231-254_GST-HsRCK_70-472_D
FEATURES             Location/Qualifiers
     source          1..6580
                     /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        1096..1114
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1115..1139
                     /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             1200..1853
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1811..1833
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1860..1883
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3117..3161
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3199..3217)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3213..3260
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3490..4305)
                     /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(3490..4305)
                     /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     4213..4232
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4306..4397)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4413..5162)
                     /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(4423..4440)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4663..4682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5358..6440)
                     /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     6402..6421
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6441..6518)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6564..6580,1..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 cccgggttcg gggatgatat tgaggagatc ccagacacag attttgattt
      181 tgaagggaac ctggctcttt ttgacaaggc agctgtgttt gaggagattg atacctatga
      241 aaggagaagt ggtggtagtg gtggtcccat tgtctatcga cggatcatag tgccccacaa
      301 cgtgagcaag gagttctgca cggactctgg cctggttgtc ccaagtattt cctatgagct
      361 gcataaaaag ctgttgtccg tggctgagaa gcatgggctg acccttgagc ggagactgga
      421 gatgacaggt gtgtgtgcca gtcagatggc actgaccctc ctcggaggac ctaacaggtt
      481 gaatcccaaa aatgttcacc agaggcctac agtggctcta ctgtgtggac ctcatgtgaa
      541 gggggctcag ggtatcagct gtggaaggca cctagccaac catgatgtcc aggtcatcct
      601 tttcctgccc aattttgtca agatgttgga atctatcacc aatgagctgt cgctcttcag
      661 caagacccaa ggccaacaag tgtctagcct caaagatctg cccactagcc ctgtggacct
      721 ggtcatcaac tgcctggatt gccctgagaa cgtcttcctg cgcgatcaac cctggtacaa
      781 ggcagctgtg gcctgggcca accagaaccg ggcaccagta ctcagcatag accctcctgt
      841 gcatgaagtc gaacagggca ttgatgccaa atggtcactg gcactgggcc tgcctctgcc
      901 actgggggag cacgcaggcc gtatctattt gtgcgacatt ggcattcccc agcaggtctt
      961 ccaggaggtg ggcatcaact accactcgcc ctttggctgc aagtttgtta tcccactgca
     1021 ctctgcttag gcggccgcat aatgcttaag tcgaacagaa agtaatcgta ttgtacacgg
     1081 ccgcataatc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
     1141 catcttagta tattagttaa gtataagaag gagatataca tatggcagat ctcaattgga
     1201 tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact cgacttcttt
     1261 tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa ggtgataaat
     1321 ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat tatattgatg
     1381 gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac aagcacaaca
     1441 tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga gcggttttgg
     1501 atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact ctcaaagttg
     1561 attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta tgtcataaaa
     1621 catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac gctcttgatg
     1681 ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa
     1741 aacgtattga agctatccca caaattgata agtacttgaa atccagcaag tatatagcat
     1801 ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca aaatcggatc
     1861 tggaagttct gttccagggg cccctgggat ccaaggatct aagaatcaaa acttcggatg
     1921 tgacctccac aaaaggaaat gagtttgaag attactgttt gaaacgggag ttactgatgg
     1981 gaatttttga aatgggctgg gaaaagccat ctcctattca ggaggagagc attcccattg
     2041 ctttatctgg tagggatatc ttagctagag caaaaaatgg aacaggcaag agcggtgcct
     2101 acctcattcc cttacttgaa cggctagacc tgaagaagga caatatacaa gcaatggtga
     2161 ttgttcccac tagagaactt gctctacagg tcagtcaaat ttgcatccag gtcagcaaac
     2221 acatgggagg ggccaaagtg atggcaacca caggaggaac caatttacga gatgacataa
     2281 tgaggcttga tgatacagtg cacgtggtga ttgctacccc tgggagaatc ctggatctta
     2341 ttaagaaagg agtagcaaag gttgatcatg tccagatgat agtattggat gaggcagata
     2401 agttgctgtc acaggatttt gtgcagataa tggaggatat tattctcacg ctacctaaaa
     2461 acaggcagat tttactatat tccgctactt tccctcttag tgtacagaag ttcatgaatt
     2521 cccatttgca gaaaccctat gagattaacc tgatggagga actaactctg aagggagtaa
     2581 cccagtacta cgcatatgta actgagcgcc aaaaagtaca ctgcctcaac acacttttct
     2641 ccaggcttca gataaaccag tcgatcattt tctgtaactc ctctcagcga gttgaattgc
     2701 tagccaagaa gatttctcaa ctgggttatt cttgcttcta tattcatgct aaaatgaggc
     2761 aggaacatcg aaatcgtgta tttcatgatt tccgaaatgg cttatgccgc aatcttgttt
     2821 gcactgatct gtttacccga ggtattgata tacaagctgt gaatgtggta ataaactttg
     2881 atttcccaaa gctggcagag acctatctcc atcgtattgg aagatcaggt cgctttggtc
     2941 atcttggctt agccatcaac ttgatcacat atgatgatcg cttcaacctg aaaagtattg
     3001 aggagcagct gggaacagaa attaaaccta ttccgagcaa cattgataag agcctgtatg
     3061 tggcagaata ccacagcgag cctgtagaag atgagaaacc ttaactcgag tctggtaaag
     3121 aaaccgctgc tgcgaaattt gaacgccagc acatggactc gtctactagc gcagcttaat
     3181 taacctaggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     3241 gggtcttgag gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacactgc
     3301 ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct
     3361 gccctgaacc gacgacaagc tgacgaccgg gtctccgcaa gtggcacttt tcggggaaat
     3421 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3481 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     3541 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     3601 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     3661 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     3721 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     3781 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     3841 tcgtgattgc gcctgagcga gacgaaatac gcggtcgctg ttaaaaggac aattacaaac
     3901 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     3961 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     4021 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     4081 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     4141 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     4201 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     4261 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcatactct tcctttttca
     4321 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4381 ttagaaaaat aaacaaatag gcatgcagcg ctcttccgct tcctcgctca ctgactcgct
     4441 acgctcggtc gttcgactgc ggcgagcggt gtcagctcac tcaaaagcgg taatacggtt
     4501 atccacagaa tcaggggata aagccggaaa gaacatgtga gcaaaaagca aagcaccgga
     4561 agaagccaac gccgcaggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4621 atcgacgctc aagccagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4681 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4741 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt tggtatctca
     4801 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4861 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4921 cgccactggc agcagccatt ggtaactgat ttagaggact ttgtcttgaa gttatgcacc
     4981 tgttaaggct aaactgaaag aacagatttt ggtgagtgcg gtcctccaac ccacttacct
     5041 tggttcaaag agttggtagc tcagcgaacc ttgagaaaac caccgttggt agcggtggtt
     5101 tttctttatt tatgagatga tgaatcaatc ggtctatcaa gtcaacgaac agctattccg
     5161 ttactctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata
     5221 cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg
     5281 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     5341 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5401 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     5461 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     5521 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     5581 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     5641 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     5701 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     5761 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     5821 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     5881 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     5941 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     6001 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     6061 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     6121 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     6181 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     6241 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     6301 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     6361 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     6421 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     6481 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     6541 atgcgactcc tgcattagga aattaatacg actcactata
//
