LOCUS       sequence-288555-        6553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsEDC3_192-508del231-263-HsRck_70-472.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6553)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6553)
  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_70-472_D
FEATURES             Location/Qualifiers
     source          1..6553
                     /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             3090..3134
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3172..3190)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3186..3233
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3463..4278)
                     /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(3463..4278)
                     /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     4186..4205
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4279..4370)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4386..5135)
                     /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(4396..4413)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4636..4655)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5331..6413)
                     /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     6375..6394
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6414..6491)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6537..6553,1..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 gatccaagga tctaagaatc aaaacttcgg atgtgacctc cacaaaagga aatgagtttg
     1921 aagattactg tttgaaacgg gagttactga tgggaatttt tgaaatgggc tgggaaaagc
     1981 catctcctat tcaggaggag agcattccca ttgctttatc tggtagggat atcttagcta
     2041 gagcaaaaaa tggaacaggc aagagcggtg cctacctcat tcccttactt gaacggctag
     2101 acctgaagaa ggacaatata caagcaatgg tgattgttcc cactagagaa cttgctctac
     2161 aggtcagtca aatttgcatc caggtcagca aacacatggg aggggccaaa gtgatggcaa
     2221 ccacaggagg aaccaattta cgagatgaca taatgaggct tgatgataca gtgcacgtgg
     2281 tgattgctac ccctgggaga atcctggatc ttattaagaa aggagtagca aaggttgatc
     2341 atgtccagat gatagtattg gatgaggcag ataagttgct gtcacaggat tttgtgcaga
     2401 taatggagga tattattctc acgctaccta aaaacaggca gattttacta tattccgcta
     2461 ctttccctct tagtgtacag aagttcatga attcccattt gcagaaaccc tatgagatta
     2521 acctgatgga ggaactaact ctgaagggag taacccagta ctacgcatat gtaactgagc
     2581 gccaaaaagt acactgcctc aacacacttt tctccaggct tcagataaac cagtcgatca
     2641 ttttctgtaa ctcctctcag cgagttgaat tgctagccaa gaagatttct caactgggtt
     2701 attcttgctt ctatattcat gctaaaatga ggcaggaaca tcgaaatcgt gtatttcatg
     2761 atttccgaaa tggcttatgc cgcaatcttg tttgcactga tctgtttacc cgaggtattg
     2821 atatacaagc tgtgaatgtg gtaataaact ttgatttccc aaagctggca gagacctatc
     2881 tccatcgtat tggaagatca ggtcgctttg gtcatcttgg cttagccatc aacttgatca
     2941 catatgatga tcgcttcaac ctgaaaagta ttgaggagca gctgggaaca gaaattaaac
     3001 ctattccgag caacattgat aagagcctgt atgtggcaga ataccacagc gagcctgtag
     3061 aagatgagaa accttaactc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc
     3121 agcacatgga ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc
     3181 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac
     3241 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
     3301 agcaatagac ataagcggct atttaacgac cctgccctga accgacgaca agctgacgac
     3361 cgggtctccg caagtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3421 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     3481 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     3541 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     3601 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     3661 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     3721 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     3781 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     3841 tacgcggtcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     3901 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     3961 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     4021 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     4081 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     4141 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     4201 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     4261 ccgttgaata tggctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4321 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggcatgca
     4381 gcgctcttcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     4441 ggtgtcagct cactcaaaag cggtaatacg gttatccaca gaatcagggg ataaagccgg
     4501 aaagaacatg tgagcaaaaa gcaaagcacc ggaagaagcc aacgccgcag gcgtttttcc
     4561 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagccag aggtggcgaa
     4621 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4681 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4741 cgctttctca tagctcacgc tgttggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4801 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4861 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc attggtaact
     4921 gatttagagg actttgtctt gaagttatgc acctgttaag gctaaactga aagaacagat
     4981 tttggtgagt gcggtcctcc aacccactta ccttggttca aagagttggt agctcagcga
     5041 accttgagaa aaccaccgtt ggtagcggtg gtttttcttt atttatgaga tgatgaatca
     5101 atcggtctat caagtcaacg aacagctatt ccgttactct agatttcagt gcaatttatc
     5161 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag
     5221 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     5281 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     5341 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5401 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     5461 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     5521 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     5581 tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg
     5641 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     5701 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     5761 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     5821 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     5881 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     5941 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     6001 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     6061 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     6121 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     6181 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     6241 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     6301 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     6361 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     6421 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     6481 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaaattaat
     6541 acgactcact ata
//
