LOCUS       sequence-333806-        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     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             101..118
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             128..145
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             152..445
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             446..469
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(470..472,473..475,476..1186)
                     /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(515..536)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(776..795)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1123..1144
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1852..1869
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1922..1940)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1936..1983
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2020..2475
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2107..2126)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2317..2338
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2567..3382)
                     /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(2567..3382)
                     /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     3290..3309
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3374..3393
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3504..4092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3993..4012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4246..4263
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4278..4417
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4343..4362)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4503..4525
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4519..4710)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5519..6601)
                     /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     6563..6582
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6602..6679)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6885..6904
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gggcagcagc catcatcatc atcatcacgg
      121 cagcggcctg gtgccgcgcg gcagcgctag catgtcggac tcagaagtca atcaagaagc
      181 taagccagag gtcaagccag aagtcaagcc tgagactcac atcaatttaa aggtgtccga
      241 tggatcttca gagatcttct tcaagatcaa aaagaccact cctttaagaa ggctgatgga
      301 agcgttcgct aaaagacagg gtaaggaaat ggactcctta agattcttgt acgacggtat
      361 tagaattcaa gctgatcaga cccctgaaga tttggacatg gaggataacg atattattga
      421 ggctcacaga gaacagattg gtggtctgga agttctgttc cagggtccaa tggtgagcaa
      481 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      541 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      601 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      661 cctgacctgg ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
      721 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
      781 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
      841 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
      901 caacgccatc agcgacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
      961 caacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1021 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1081 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1141 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggta ccatggcctc
     1201 cggtaacgcg cgcatcggaa agccagcccc tgacttcaag gccacagcgg tggttgatgg
     1261 cgccttcaaa gaggtgaagc tgtcggacta caaagggaag tacgtggtcc tctttttcta
     1321 ccctctggac ttcacttttg tgtgccccac cgagatcatc gcgttcagca accgtgcaga
     1381 ggacttccgc aagctgggct gtgaagtgct gggcgtctcg gtggactctc agttcaccca
     1441 cctggcttgg atcaacaccc cccggaaaga gggaggcttg ggccccctga acatccccct
     1501 gcttgctgac gtgaccagac gcttgtctga ggattacggc gtgctgaaaa cagatgaggg
     1561 cattgcctac aggggcctct ttatcatcga tggcaagggt gtccttcgcc agatcactgt
     1621 taatgatttg cctgtgggac gctccgtgga tgaggctctg cggctggtcc aggccttcca
     1681 gtacacagac gagcatgggg aagtttgtcc cgctggctgg aagcctggca gtgacacgat
     1741 taagcccaac gtggatgaca gcaaggaata tttctccaaa cacaattaaa gacaagctta
     1801 ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttagtcga gcaccaccac
     1861 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1921 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1981 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
     2041 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2101 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2161 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2221 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2281 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2341 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2401 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2461 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2521 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat
     2581 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     2641 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     2701 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     2761 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     2821 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     2881 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     2941 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     3001 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     3061 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     3121 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     3181 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     3241 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     3301 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     3361 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     3421 ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3481 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3541 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3601 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3661 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3721 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3781 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3841 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3901 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3961 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4021 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4081 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4141 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4201 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4261 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4321 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     4381 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     4441 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4501 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4561 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4621 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4681 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     4741 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     4801 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     4861 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     4921 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     4981 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     5041 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     5101 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     5161 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     5221 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     5281 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     5341 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     5401 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     5461 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     5521 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5581 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     5641 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     5701 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     5761 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     5821 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     5881 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     5941 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     6001 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     6061 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     6121 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     6181 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     6241 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     6301 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     6361 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     6421 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     6481 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     6541 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     6601 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     6661 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     6721 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     6781 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     6841 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     6901 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     6961 aggatcgaga tctcgatccc gcgaaat
//
