LOCUS       sequence-288620-        4717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmEDC3_4-70.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4717)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 4717)
  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: pRSFDuetG-DmEDC3_4-70 (MCSII)_E
FEATURES             Location/Qualifiers
     source          1..4717
                     /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        244..262
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    263..287
                     /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             333..986
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     944..966
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             993..1016
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1254..1298
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1336..1354)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1350..1397
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1627..2442)
                     /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(1627..2442)
                     /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     2350..2369
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2443..2534)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2550..3299)
                     /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(2560..2577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2800..2819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3495..4577)
                     /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     4539..4558
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4578..4655)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4701..4717,1..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 ccccaggatc cgaattcgag ctcggcgcgc ctgcaggtcg acaagcttgc
      181 ggccgcataa tgcttaagtc gaacagaaag taatcgtatt gtacacggcc gcataatcga
      241 aattaatacg actcactata ggggaattgt gagcggataa caattcccca tcttagtata
      301 ttagttaagt ataagaagga gatatacata atatgtcccc tatactaggt tattggaaaa
      361 ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag
      421 agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt
      481 tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg
      541 ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg
      601 cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg
      661 catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc
      721 tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc
      781 atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc
      841 tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg
      901 ataagtactt gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt
      961 ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt tctgttccag gggccccata
     1021 tggcagatct cacggatcaa gactggatcg gctgtgcagt gtctatagcc tgcgacgagg
     1081 tcctgggcgt cttccagggc ctgatcaagc agatttctgc cgaggagatc accattgtga
     1141 gggctttccg caatggcgtg ccgctgcgaa agcagaacgc cgaggtggtc ctcaagtgca
     1201 cggacattcg cagcatcgat ctcatcgagc cctagggtac cctcgagtct ggtaaagaaa
     1261 ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc tactagcgca gcttaattaa
     1321 cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1381 tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc
     1441 cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc
     1501 ctgaaccgac gacaagctga cgaccgggtc tccgcaagtg gcacttttcg gggaaatgtg
     1561 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat
     1621 taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     1681 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     1741 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     1801 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     1861 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     1921 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     1981 tgattgcgcc tgagcgagac gaaatacgcg gtcgctgtta aaaggacaat tacaaacagg
     2041 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     2101 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     2161 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     2221 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     2281 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     2341 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     2401 tcgcggccta gagcaagacg tttcccgttg aatatggctc atactcttcc tttttcaata
     2461 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2521 gaaaaataaa caaataggca tgcagcgctc ttccgcttcc tcgctcactg actcgctacg
     2581 ctcggtcgtt cgactgcggc gagcggtgtc agctcactca aaagcggtaa tacggttatc
     2641 cacagaatca ggggataaag ccggaaagaa catgtgagca aaaagcaaag caccggaaga
     2701 agccaacgcc gcaggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2761 gacgctcaag ccagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2821 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2881 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgttgg tatctcagtt
     2941 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3001 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3061 cactggcagc agccattggt aactgattta gaggactttg tcttgaagtt atgcacctgt
     3121 taaggctaaa ctgaaagaac agattttggt gagtgcggtc ctccaaccca cttaccttgg
     3181 ttcaaagagt tggtagctca gcgaaccttg agaaaaccac cgttggtagc ggtggttttt
     3241 ctttatttat gagatgatga atcaatcggt ctatcaagtc aacgaacagc tattccgtta
     3301 ctctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
     3361 tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag ctgactgggt
     3421 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     3481 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3541 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     3601 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     3661 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     3721 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc
     3781 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     3841 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     3901 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     3961 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     4021 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     4081 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     4141 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     4201 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     4261 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     4321 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     4381 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     4441 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     4501 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     4561 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     4621 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     4681 cgactcctgc attaggaaat taatacgact cactata
//
