LOCUS       sequence-328167-        7617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded fluorescent H2O2 sensor roGFP2-Prx2 for 
            cytosolic expression in mammalian cells (for stable cell line 
            generation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7617)
  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 7617)
  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: pLPCX roGFP2-Prx2
FEATURES             Location/Qualifiers
     source          1..7617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             414..589
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    652..851
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1052..1468
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1398..1420
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1436..1452
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1566..2165
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(1566..1585)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2062..2082
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     enhancer        2340..2643
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2644..2847
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2798..2818
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(2898..2900,2901..2903,2904..3614)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2943..2964)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3204..3223)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3551..3572
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4356..4949
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5052..5074)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    5202..5235
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5295..5328
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     5348..5367
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5448..5465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5619..6207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5699..5718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6378..7169,7170..7238))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7001..7020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7239..7343)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7411..7429
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
      121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
      181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
      301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
      421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1441 tccagccctc actccttctc taggcgccgg aattgatccc ccgggctgca ggtcggccgc
     1501 cacgaccggt gccgccacca tcccctgacc cacgcccctg acccctcaca aggagacgac
     1561 cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
     1621 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
     1681 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     1741 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     1801 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     1861 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     1921 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     1981 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     2041 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     2101 ccgccgacgt cgagtgcccg aaggaccgcg cgacctggtg catgacccgc aagcccggtg
     2161 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatggct
     2221 ccgaccgaag ccgacccggg cggccccgcc gaccccgcac ccgcccccga ggcccaccga
     2281 ctctagtaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     2341 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     2401 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     2461 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     2521 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     2581 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     2641 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     2701 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     2761 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     2821 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
     2881 cggactcaga tctcgagatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2941 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     3001 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctccaccacc ggcaagctgc
     3061 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     3121 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3181 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3241 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3301 gcaacatcct ggggcacaag ctggagtaca actacaactg ccacaacgtc tatatcatgg
     3361 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     3421 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3481 tgctgcccga caaccactac ctgagcacct gctccgccct gagcaaagac cccaacgaga
     3541 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3601 acgagctgta caagactagt ggtggttcag gtggtggtgg ttcaggtggt ggtggttcag
     3661 gtggaggagg atcaggagga ggaggatcag gaggaggagg atcaggagga gaattcgcct
     3721 ccggtaacgc gcgcatcgga aagccagccc ctgacttcaa ggccacagcg gtggttgatg
     3781 gcgccttcaa agaggtgaag ctgtcggact acaaagggaa gtacgtggtc ctctttttct
     3841 accctctgga cttcactttt gtgtgcccca ccgagatcat cgcgttcagc aaccgtgcag
     3901 aggacttccg caagctgggc tgtgaagtgc tgggcgtctc ggtggactct cagttcaccc
     3961 acctggcttg gatcaacacc ccccggaaag agggaggctt gggccccctg aacatccccc
     4021 tgcttgctga cgtgaccaga cgcttgtctg aggattacgg cgtgctgaaa acagatgagg
     4081 gcattgccta caggggcctc tttatcatcg atggcaaggg tgtccttcgc cagatcactg
     4141 ttaatgattt gcctgtggga cgctccgtgg atgaggctct gcggctggtc caggccttcc
     4201 agtacacaga cgagcatggg gaagtttgtc ccgctggctg gaagcctggc agtgacacga
     4261 ttaagcccaa cgtggatgac agcaaggaat atttctccaa acacaattag atcgataaaa
     4321 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     4381 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata
     4441 gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata
     4501 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
     4561 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     4621 ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
     4681 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     4741 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     4801 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
     4861 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     4921 tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg
     4981 cccagggacc accgacccac caccgggagg taagctggct gcctcgcgcg tttcggtgat
     5041 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     5101 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     5161 gcagccatga cccagtcacg tagcgatagc ggagtgtact gataacttcg tataatgtat
     5221 gctatacgaa gttattaggt ctgaagagga gtttacgtcc agccaagcta gggccgcgat
     5281 ccggaaccct taatataact tcgtataatg tatgctatac gaagttatca gtactggctt
     5341 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     5401 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     5461 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5521 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5581 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5641 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5701 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5761 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5821 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5881 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5941 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6001 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6061 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6121 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6181 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6241 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6301 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6361 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6421 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6481 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6541 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6601 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6661 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
     6721 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6781 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6841 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6901 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6961 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     7021 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7081 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7141 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7201 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7261 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7321 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     7381 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
     7441 caagaattgc tagcaattgc tagcaattgc tagcaattca taccagatca ccgaaaactg
     7501 tcctccaaat gtgtccccct cacactccca aattcgcggg cttctgcctc ttagaccact
     7561 ctaccctatt ccccacactc accggagcca aagccgcggc ccttccgttt ctttgct
//
