LOCUS       sequence-288554-        6760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsEDC3_192-508del231-263-HsRck.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6760)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6760)
  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_192-508del231-263_GST-HsRCK_D
FEATURES             Location/Qualifiers
     source          1..6760
                     /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        1069..1087
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1088..1112
                     /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             1173..1826
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1784..1806
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1833..1856
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3297..3341
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3379..3397)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3393..3440
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3670..4485)
                     /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(3670..4485)
                     /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     4393..4412
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4486..4577)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4593..5342)
                     /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(4603..4620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4843..4862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5538..6620)
                     /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     6582..6601
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6621..6698)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6744..6760,1..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 gtggtcccca caacgtgagc aaggagttct gcacggactc
      301 tggcctggtt gtcccaagta tttcctatga gctgcataaa aagctgttgt ccgtggctga
      361 gaagcatggg ctgacccttg agcggagact ggagatgaca ggtgtgtgtg ccagtcagat
      421 ggcactgacc ctcctcggag gacctaacag gttgaatccc aaaaatgttc accagaggcc
      481 tacagtggct ctactgtgtg gacctcatgt gaagggggct cagggtatca gctgtggaag
      541 gcacctagcc aaccatgatg tccaggtcat ccttttcctg cccaattttg tcaagatgtt
      601 ggaatctatc accaatgagc tgtcgctctt cagcaagacc caaggccaac aagtgtctag
      661 cctcaaagat ctgcccacta gccctgtgga cctggtcatc aactgcctgg attgccctga
      721 gaacgtcttc ctgcgcgatc aaccctggta caaggcagct gtggcctggg ccaaccagaa
      781 ccgggcacca gtactcagca tagaccctcc tgtgcatgaa gtcgaacagg gcattgatgc
      841 caaatggtca ctggcactgg gcctgcctct gccactgggg gagcacgcag gccgtatcta
      901 tttgtgcgac attggcattc cccagcaggt cttccaggag gtgggcatca actaccactc
      961 gccctttggc tgcaagtttg ttatcccact gcactctgct taggcggccg cataatgctt
     1021 aagtcgaaca gaaagtaatc gtattgtaca cggccgcata atcgaaatta atacgactca
     1081 ctatagggga attgtgagcg gataacaatt ccccatctta gtatattagt taagtataag
     1141 aaggagatat acatatggca gatctcaatt ggatgtcccc tatactaggt tattggaaaa
     1201 ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag
     1261 agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt
     1321 tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg
     1381 ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg
     1441 cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg
     1501 catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc
     1561 tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc
     1621 atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc
     1681 tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg
     1741 ataagtactt gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt
     1801 ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt tctgttccag gggcccctgg
     1861 gatccatggg tctgtccagt caaaatggtc agctgagagg ccctgtgaaa cccactggtg
     1921 gccctggagg agggggcaca cagacacagc aacagatgaa ccagctgaaa aacaccaaca
     1981 caatcaataa tggcactcag cagcaagcac agagtatgac caccactatt aaacctggtg
     2041 atgactggaa aaagacttta aaactccctc caaaggatct aagaatcaaa acttcggatg
     2101 tgacctccac aaaaggaaat gagtttgaag attactgttt gaaacgggag ttactgatgg
     2161 gaatttttga aatgggctgg gaaaagccat ctcctattca ggaggagagc attcccattg
     2221 ctttatctgg tagggatatc ttagctagag caaaaaatgg aacaggcaag agcggtgcct
     2281 acctcattcc cttacttgaa cggctagacc tgaagaagga caatatacaa gcaatggtga
     2341 ttgttcccac tagagaactt gctctacagg tcagtcaaat ttgcatccag gtcagcaaac
     2401 acatgggagg ggccaaagtg atggcaacca caggaggaac caatttacga gatgacataa
     2461 tgaggcttga tgatacagtg cacgtggtga ttgctacccc tgggagaatc ctggatctta
     2521 ttaagaaagg agtagcaaag gttgatcatg tccagatgat agtattggat gaggcagata
     2581 agttgctgtc acaggatttt gtgcagataa tggaggatat tattctcacg ctacctaaaa
     2641 acaggcagat tttactatat tccgctactt tccctcttag tgtacagaag ttcatgaatt
     2701 cccatttgca gaaaccctat gagattaacc tgatggagga actaactctg aagggagtaa
     2761 cccagtacta cgcatatgta actgagcgcc aaaaagtaca ctgcctcaac acacttttct
     2821 ccaggcttca gataaaccag tcgatcattt tctgtaactc ctctcagcga gttgaattgc
     2881 tagccaagaa gatttctcaa ctgggttatt cttgcttcta tattcatgct aaaatgaggc
     2941 aggaacatcg aaatcgtgta tttcatgatt tccgaaatgg cttatgccgc aatcttgttt
     3001 gcactgatct gtttacccga ggtattgata tacaagctgt gaatgtggta ataaactttg
     3061 atttcccaaa gctggcagag acctatctcc atcgtattgg aagatcaggt cgctttggtc
     3121 atcttggctt agccatcaac ttgatcacat atgatgatcg cttcaacctg aaaagtattg
     3181 aggagcagct gggaacagaa attaaaccta ttccgagcaa cattgataag agcctgtatg
     3241 tggcagaata ccacagcgag cctgtagaag atgagaaacc ttaactcgag tctggtaaag
     3301 aaaccgctgc tgcgaaattt gaacgccagc acatggactc gtctactagc gcagcttaat
     3361 taacctaggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     3421 gggtcttgag gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacactgc
     3481 ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct
     3541 gccctgaacc gacgacaagc tgacgaccgg gtctccgcaa gtggcacttt tcggggaaat
     3601 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3661 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     3721 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     3781 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     3841 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     3901 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     3961 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     4021 tcgtgattgc gcctgagcga gacgaaatac gcggtcgctg ttaaaaggac aattacaaac
     4081 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     4141 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     4201 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     4261 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     4321 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     4381 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     4441 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcatactct tcctttttca
     4501 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4561 ttagaaaaat aaacaaatag gcatgcagcg ctcttccgct tcctcgctca ctgactcgct
     4621 acgctcggtc gttcgactgc ggcgagcggt gtcagctcac tcaaaagcgg taatacggtt
     4681 atccacagaa tcaggggata aagccggaaa gaacatgtga gcaaaaagca aagcaccgga
     4741 agaagccaac gccgcaggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4801 atcgacgctc aagccagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4861 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4921 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt tggtatctca
     4981 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5041 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5101 cgccactggc agcagccatt ggtaactgat ttagaggact ttgtcttgaa gttatgcacc
     5161 tgttaaggct aaactgaaag aacagatttt ggtgagtgcg gtcctccaac ccacttacct
     5221 tggttcaaag agttggtagc tcagcgaacc ttgagaaaac caccgttggt agcggtggtt
     5281 tttctttatt tatgagatga tgaatcaatc ggtctatcaa gtcaacgaac agctattccg
     5341 ttactctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata
     5401 cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg
     5461 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     5521 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5581 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     5641 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     5701 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     5761 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     5821 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     5881 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     5941 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     6001 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     6061 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     6121 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     6181 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     6241 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     6301 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     6361 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     6421 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     6481 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     6541 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     6601 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     6661 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     6721 atgcgactcc tgcattagga aattaatacg actcactata
//
