LOCUS       sequence-288557-        6787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsEDC3_192-508del231-254-HsRck.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6787)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6787)
  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_D
FEATURES             Location/Qualifiers
     source          1..6787
                     /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             3324..3368
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3406..3424)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3420..3467
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3697..4512)
                     /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(3697..4512)
                     /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     4420..4439
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4513..4604)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4620..5369)
                     /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(4630..4647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4870..4889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5565..6647)
                     /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     6609..6628
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6648..6725)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6771..6787,1..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 ccatgggtct gtccagtcaa aatggtcagc
     1921 tgagaggccc tgtgaaaccc actggtggcc ctggaggagg gggcacacag acacagcaac
     1981 agatgaacca gctgaaaaac accaacacaa tcaataatgg cactcagcag caagcacaga
     2041 gtatgaccac cactattaaa cctggtgatg actggaaaaa gactttaaaa ctccctccaa
     2101 aggatctaag aatcaaaact tcggatgtga cctccacaaa aggaaatgag tttgaagatt
     2161 actgtttgaa acgggagtta ctgatgggaa tttttgaaat gggctgggaa aagccatctc
     2221 ctattcagga ggagagcatt cccattgctt tatctggtag ggatatctta gctagagcaa
     2281 aaaatggaac aggcaagagc ggtgcctacc tcattccctt acttgaacgg ctagacctga
     2341 agaaggacaa tatacaagca atggtgattg ttcccactag agaacttgct ctacaggtca
     2401 gtcaaatttg catccaggtc agcaaacaca tgggaggggc caaagtgatg gcaaccacag
     2461 gaggaaccaa tttacgagat gacataatga ggcttgatga tacagtgcac gtggtgattg
     2521 ctacccctgg gagaatcctg gatcttatta agaaaggagt agcaaaggtt gatcatgtcc
     2581 agatgatagt attggatgag gcagataagt tgctgtcaca ggattttgtg cagataatgg
     2641 aggatattat tctcacgcta cctaaaaaca ggcagatttt actatattcc gctactttcc
     2701 ctcttagtgt acagaagttc atgaattccc atttgcagaa accctatgag attaacctga
     2761 tggaggaact aactctgaag ggagtaaccc agtactacgc atatgtaact gagcgccaaa
     2821 aagtacactg cctcaacaca cttttctcca ggcttcagat aaaccagtcg atcattttct
     2881 gtaactcctc tcagcgagtt gaattgctag ccaagaagat ttctcaactg ggttattctt
     2941 gcttctatat tcatgctaaa atgaggcagg aacatcgaaa tcgtgtattt catgatttcc
     3001 gaaatggctt atgccgcaat cttgtttgca ctgatctgtt tacccgaggt attgatatac
     3061 aagctgtgaa tgtggtaata aactttgatt tcccaaagct ggcagagacc tatctccatc
     3121 gtattggaag atcaggtcgc tttggtcatc ttggcttagc catcaacttg atcacatatg
     3181 atgatcgctt caacctgaaa agtattgagg agcagctggg aacagaaatt aaacctattc
     3241 cgagcaacat tgataagagc ctgtatgtgg cagaatacca cagcgagcct gtagaagatg
     3301 agaaacctta actcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca
     3361 tggactcgtc tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac
     3421 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg
     3481 catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat
     3541 agacataagc ggctatttaa cgaccctgcc ctgaaccgac gacaagctga cgaccgggtc
     3601 tccgcaagtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3661 atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa
     3721 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     3781 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     3841 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     3901 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     3961 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     4021 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     4081 gtcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     4141 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     4201 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     4261 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     4321 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     4381 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     4441 atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg
     4501 aatatggctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4561 catgagcgga tacatatttg aatgtattta gaaaaataaa caaataggca tgcagcgctc
     4621 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggtgtc
     4681 agctcactca aaagcggtaa tacggttatc cacagaatca ggggataaag ccggaaagaa
     4741 catgtgagca aaaagcaaag caccggaaga agccaacgcc gcaggcgttt ttccataggc
     4801 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag ccagaggtgg cgaaacccga
     4861 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4921 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4981 ctcatagctc acgctgttgg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5041 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5101 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccattggt aactgattta
     5161 gaggactttg tcttgaagtt atgcacctgt taaggctaaa ctgaaagaac agattttggt
     5221 gagtgcggtc ctccaaccca cttaccttgg ttcaaagagt tggtagctca gcgaaccttg
     5281 agaaaaccac cgttggtagc ggtggttttt ctttatttat gagatgatga atcaatcggt
     5341 ctatcaagtc aacgaacagc tattccgtta ctctagattt cagtgcaatt tatctcttca
     5401 aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc
     5461 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
     5521 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     5581 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5641 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     5701 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     5761 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     5821 gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg
     5881 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     5941 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     6001 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     6061 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     6121 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     6181 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     6241 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     6301 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     6361 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     6421 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     6481 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     6541 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     6601 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     6661 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     6721 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact
     6781 cactata
//
