LOCUS       sequence-288556-        5875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsEDC3_192-508del231-263-HsRck_296-472.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5875)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5875)
  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_296-472_D
FEATURES             Location/Qualifiers
     source          1..5875
                     /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             2412..2456
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2494..2512)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2508..2555
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2785..3600)
                     /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(2785..3600)
                     /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     3508..3527
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3601..3692)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3708..4457)
                     /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(3718..3735)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3958..3977)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4653..5735)
                     /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     5697..5716
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5736..5813)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5859..5875,1..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 gatccaaggg agtaacccag tactacgcat atgtaactga gcgccaaaaa gtacactgcc
     1921 tcaacacact tttctccagg cttcagataa accagtcgat cattttctgt aactcctctc
     1981 agcgagttga attgctagcc aagaagattt ctcaactggg ttattcttgc ttctatattc
     2041 atgctaaaat gaggcaggaa catcgaaatc gtgtatttca tgatttccga aatggcttat
     2101 gccgcaatct tgtttgcact gatctgttta cccgaggtat tgatatacaa gctgtgaatg
     2161 tggtaataaa ctttgatttc ccaaagctgg cagagaccta tctccatcgt attggaagat
     2221 caggtcgctt tggtcatctt ggcttagcca tcaacttgat cacatatgat gatcgcttca
     2281 acctgaaaag tattgaggag cagctgggaa cagaaattaa acctattccg agcaacattg
     2341 ataagagcct gtatgtggca gaataccaca gcgagcctgt agaagatgag aaaccttaac
     2401 tcgagtctgg taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgtcta
     2461 ctagcgcagc ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc
     2521 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc
     2581 acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg
     2641 ctatttaacg accctgccct gaaccgacga caagctgacg accgggtctc cgcaagtggc
     2701 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2761 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     2821 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     2881 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     2941 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     3001 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     3061 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     3121 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcggt cgctgttaaa
     3181 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     3241 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     3301 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     3361 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     3421 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     3481 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     3541 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     3601 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3661 catatttgaa tgtatttaga aaaataaaca aataggcatg cagcgctctt ccgcttcctc
     3721 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggtgtcag ctcactcaaa
     3781 agcggtaata cggttatcca cagaatcagg ggataaagcc ggaaagaaca tgtgagcaaa
     3841 aagcaaagca ccggaagaag ccaacgccgc aggcgttttt ccataggctc cgcccccctg
     3901 acgagcatca caaaaatcga cgctcaagcc agaggtggcg aaacccgaca ggactataaa
     3961 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4021 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4081 gctgttggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4141 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4201 taagacacga cttatcgcca ctggcagcag ccattggtaa ctgatttaga ggactttgtc
     4261 ttgaagttat gcacctgtta aggctaaact gaaagaacag attttggtga gtgcggtcct
     4321 ccaacccact taccttggtt caaagagttg gtagctcagc gaaccttgag aaaaccaccg
     4381 ttggtagcgg tggtttttct ttatttatga gatgatgaat caatcggtct atcaagtcaa
     4441 cgaacagcta ttccgttact ctagatttca gtgcaattta tctcttcaaa tgtagcacct
     4501 gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc
     4561 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     4621 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4681 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4741 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     4801 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     4861 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     4921 cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     4981 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     5041 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     5101 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     5161 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     5221 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     5281 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     5341 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     5401 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     5461 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     5521 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     5581 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     5641 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     5701 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     5761 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     5821 tcgacgctct cccttatgcg actcctgcat taggaaatta atacgactca ctata
//
