LOCUS       sequence-305307-        6987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded HomoFRET-based probe to monitor peroxiredoxin2 
            (Prx2) oligomerization (Recombinant bacterial expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6987)
  AUTHORS   Pastor-Flores D, Talwar D, Pedre B, Dick TP
  TITLE     Real-time monitoring of peroxiredoxin oligomerization dynamics in 
            living cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Jun 29. pii: 1915275117. doi: 
            10.1073/pnas.1915275117.
  PUBMED    32601209
REFERENCE   2  (bases 1 to 6987)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2020 Jun 29. pii: 1915275117. doi: 
            10.1073/pnas.1915275117."
COMMENT     Sequence Label: pET-SUMO mCER-Prx2
FEATURES             Location/Qualifiers
     source          1..6987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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)."
     RBS             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             309..326
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             336..353
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             360..653
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             654..677
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(678..680,681..683,684..1394)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(723..744)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(984..1003)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1331..1352
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2060..2077
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2130..2148)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2144..2191
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2228..2683
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2315..2334)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2525..2546
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2775..3590)
                     /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(2775..3590)
                     /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     3498..3517
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3582..3601
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3712..4300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4201..4220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4454..4471
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4486..4625
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4551..4570)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4711..4733
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4727..4918)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5727..6809)
                     /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     6771..6790
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6810..6887)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat atacatatgg
      301 gcagcagcca tcatcatcat catcacggca gcggcctggt gccgcgcggc agcgctagca
      361 tgtcggactc agaagtcaat caagaagcta agccagaggt caagccagaa gtcaagcctg
      421 agactcacat caatttaaag gtgtccgatg gatcttcaga gatcttcttc aagatcaaaa
      481 agaccactcc tttaagaagg ctgatggaag cgttcgctaa aagacagggt aaggaaatgg
      541 actccttaag attcttgtac gacggtatta gaattcaagc tgatcagacc cctgaagatt
      601 tggacatgga ggataacgat attattgagg ctcacagaga acagattggt ggtctggaag
      661 ttctgttcca gggtccaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      721 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      781 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      841 ccgtgccctg gcccaccctc gtgaccaccc tgacctgggg cgtgcagtgc ttcgcccgct
      901 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      961 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1021 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1081 gcaacatcct ggggcacaag ctggagtaca acgccatcag cgacaacgtc tatatcaccg
     1141 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     1201 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1261 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
     1321 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1381 acgagctgta caagggtacc atggcctccg gtaacgcgcg catcggaaag ccagcccctg
     1441 acttcaaggc cacagcggtg gttgatggcg ccttcaaaga ggtgaagctg tcggactaca
     1501 aagggaagta cgtggtcctc tttttctacc ctctggactt cacttttgtg tgccccaccg
     1561 agatcatcgc gttcagcaac cgtgcagagg acttccgcaa gctgggctgt gaagtgctgg
     1621 gcgtctcggt ggactctcag ttcacccacc tggcttggat caacaccccc cggaaagagg
     1681 gaggcttggg ccccctgaac atccccctgc ttgctgacgt gaccagacgc ttgtctgagg
     1741 attacggcgt gctgaaaaca gatgagggca ttgcctacag gggcctcttt atcatcgatg
     1801 gcaagggtgt ccttcgccag atcactgtta atgatttgcc tgtgggacgc tccgtggatg
     1861 aggctctgcg gctggtccag gccttccagt acacagacga gcatggggaa gtttgtcccg
     1921 ctggctggaa gcctggcagt gacacgatta agcccaacgt ggatgacagc aaggaatatt
     1981 tctccaaaca caattaaaga caagcttagg tatttattcg gcgcaaagtg cgtcgggtga
     2041 tgctgccaac ttagtcgagc accaccacca ccaccactga gatccggctg ctaacaaagc
     2101 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     2161 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
     2221 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2281 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2341 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2401 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2461 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2521 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2581 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2641 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     2701 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2761 tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     2821 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     2881 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     2941 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     3001 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
     3061 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     3121 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     3181 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     3241 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     3301 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     3361 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     3421 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     3481 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     3541 gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     3601 gtattactgt ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga
     3661 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3721 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3781 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3841 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3901 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3961 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4021 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4081 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4141 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4201 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4261 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4321 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4381 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4441 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     4501 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     4561 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     4621 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4681 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4741 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     4801 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     4861 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     4921 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     4981 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     5041 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     5101 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     5161 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     5221 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     5281 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     5341 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg
     5401 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     5461 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     5521 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     5581 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     5641 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     5701 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5761 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     5821 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     5881 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     5941 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     6001 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     6061 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     6121 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     6181 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     6241 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     6301 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     6361 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     6421 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     6481 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     6541 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     6601 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     6661 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     6721 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     6781 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     6841 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     6901 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     6961 caccgccgcc gcaaggaatg gtgcatg
//
