LOCUS       sequence-333993-        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"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..150
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        1113..1131
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1132..1156
                     /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             1217..1870
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1828..1850
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1877..1900
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3134..3178
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3216..3234)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3230..3277
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3507..4322)
                     /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(3507..4322)
                     /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     4230..4249
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4323..4414)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4430..5179)
                     /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(4440..4457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4680..4699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5375..6457)
                     /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     6419..6438
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6458..6535)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcaccatca tcaccacagc
      121 gcgggtctgg aagttctgtt ccaggggccc gggttcgggg atgatattga ggagatccca
      181 gacacagatt ttgattttga agggaacctg gctctttttg acaaggcagc tgtgtttgag
      241 gagattgata cctatgaaag gagaagtggt ggtagtggtg gtcccattgt ctatcgacgg
      301 atcatagtgc cccacaacgt gagcaaggag ttctgcacgg actctggcct ggttgtccca
      361 agtatttcct atgagctgca taaaaagctg ttgtccgtgg ctgagaagca tgggctgacc
      421 cttgagcgga gactggagat gacaggtgtg tgtgccagtc agatggcact gaccctcctc
      481 ggaggaccta acaggttgaa tcccaaaaat gttcaccaga ggcctacagt ggctctactg
      541 tgtggacctc atgtgaaggg ggctcagggt atcagctgtg gaaggcacct agccaaccat
      601 gatgtccagg tcatcctttt cctgcccaat tttgtcaaga tgttggaatc tatcaccaat
      661 gagctgtcgc tcttcagcaa gacccaaggc caacaagtgt ctagcctcaa agatctgccc
      721 actagccctg tggacctggt catcaactgc ctggattgcc ctgagaacgt cttcctgcgc
      781 gatcaaccct ggtacaaggc agctgtggcc tgggccaacc agaaccgggc accagtactc
      841 agcatagacc ctcctgtgca tgaagtcgaa cagggcattg atgccaaatg gtcactggca
      901 ctgggcctgc ctctgccact gggggagcac gcaggccgta tctatttgtg cgacattggc
      961 attccccagc aggtcttcca ggaggtgggc atcaactacc actcgccctt tggctgcaag
     1021 tttgttatcc cactgcactc tgcttaggcg gccgcataat gcttaagtcg aacagaaagt
     1081 aatcgtattg tacacggccg cataatcgaa attaatacga ctcactatag gggaattgtg
     1141 agcggataac aattccccat cttagtatat tagttaagta taagaaggag atatacatat
     1201 ggcagatctc aattggatgt cccctatact aggttattgg aaaattaagg gccttgtgca
     1261 acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg
     1321 cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct
     1381 tccttattat attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat
     1441 agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct
     1501 tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt
     1561 tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga
     1621 tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt
     1681 gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa
     1741 attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc
     1801 cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca
     1861 tcctccaaaa tcggatctgg aagttctgtt ccaggggccc ctgggatcca aggatctaag
     1921 aatcaaaact tcggatgtga cctccacaaa aggaaatgag tttgaagatt actgtttgaa
     1981 acgggagtta ctgatgggaa tttttgaaat gggctgggaa aagccatctc ctattcagga
     2041 ggagagcatt cccattgctt tatctggtag ggatatctta gctagagcaa aaaatggaac
     2101 aggcaagagc ggtgcctacc tcattccctt acttgaacgg ctagacctga agaaggacaa
     2161 tatacaagca atggtgattg ttcccactag agaacttgct ctacaggtca gtcaaatttg
     2221 catccaggtc agcaaacaca tgggaggggc caaagtgatg gcaaccacag gaggaaccaa
     2281 tttacgagat gacataatga ggcttgatga tacagtgcac gtggtgattg ctacccctgg
     2341 gagaatcctg gatcttatta agaaaggagt agcaaaggtt gatcatgtcc agatgatagt
     2401 attggatgag gcagataagt tgctgtcaca ggattttgtg cagataatgg aggatattat
     2461 tctcacgcta cctaaaaaca ggcagatttt actatattcc gctactttcc ctcttagtgt
     2521 acagaagttc atgaattccc atttgcagaa accctatgag attaacctga tggaggaact
     2581 aactctgaag ggagtaaccc agtactacgc atatgtaact gagcgccaaa aagtacactg
     2641 cctcaacaca cttttctcca ggcttcagat aaaccagtcg atcattttct gtaactcctc
     2701 tcagcgagtt gaattgctag ccaagaagat ttctcaactg ggttattctt gcttctatat
     2761 tcatgctaaa atgaggcagg aacatcgaaa tcgtgtattt catgatttcc gaaatggctt
     2821 atgccgcaat cttgtttgca ctgatctgtt tacccgaggt attgatatac aagctgtgaa
     2881 tgtggtaata aactttgatt tcccaaagct ggcagagacc tatctccatc gtattggaag
     2941 atcaggtcgc tttggtcatc ttggcttagc catcaacttg atcacatatg atgatcgctt
     3001 caacctgaaa agtattgagg agcagctggg aacagaaatt aaacctattc cgagcaacat
     3061 tgataagagc ctgtatgtgg cagaatacca cagcgagcct gtagaagatg agaaacctta
     3121 actcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc
     3181 tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac tagcataacc
     3241 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa
     3301 gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc
     3361 ggctatttaa cgaccctgcc ctgaaccgac gacaagctga cgaccgggtc tccgcaagtg
     3421 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3481 atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc
     3541 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     3601 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     3661 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     3721 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
     3781 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     3841 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg gtcgctgtta
     3901 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     3961 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     4021 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     4081 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     4141 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     4201 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     4261 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc
     4321 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4381 tacatatttg aatgtattta gaaaaataaa caaataggca tgcagcgctc ttccgcttcc
     4441 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggtgtc agctcactca
     4501 aaagcggtaa tacggttatc cacagaatca ggggataaag ccggaaagaa catgtgagca
     4561 aaaagcaaag caccggaaga agccaacgcc gcaggcgttt ttccataggc tccgcccccc
     4621 tgacgagcat cacaaaaatc gacgctcaag ccagaggtgg cgaaacccga caggactata
     4681 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4741 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4801 acgctgttgg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4861 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4921 ggtaagacac gacttatcgc cactggcagc agccattggt aactgattta gaggactttg
     4981 tcttgaagtt atgcacctgt taaggctaaa ctgaaagaac agattttggt gagtgcggtc
     5041 ctccaaccca cttaccttgg ttcaaagagt tggtagctca gcgaaccttg agaaaaccac
     5101 cgttggtagc ggtggttttt ctttatttat gagatgatga atcaatcggt ctatcaagtc
     5161 aacgaacagc tattccgtta ctctagattt cagtgcaatt tatctcttca aatgtagcac
     5221 ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca
     5281 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     5341 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5401 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5461 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     5521 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     5581 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     5641 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     5701 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     5761 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     5821 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     5881 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     5941 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     6001 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     6061 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     6121 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     6181 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     6241 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     6301 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     6361 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     6421 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     6481 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     6541 tctcgacgct ctcccttatg cgactcctgc attaggaaat
//
