LOCUS       sequence-305308-        7546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded HomoFRET-based probe to monitor peroxiredoxin2 
            (Prx2) oligomerization (for stable cell line generation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7546)
  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 7546)
  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 mCER-Prx2
FEATURES             Location/Qualifiers
     source          1..7546
                     /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(2901..2903,2904..2906,2907..3617)
                     /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(2946..2967)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3207..3226)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3554..3575
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4285..4878
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4981..5003)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    5131..5164
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5224..5257
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     5277..5296
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5377..5394)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5548..6136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5628..5647)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6307..7098,7099..7167))
                     /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     6930..6949
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7168..7272)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7340..7358
                     /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 tctcgagacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2941 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3001 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3061 tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag tgcttcgccc
     3121 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3181 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3241 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3301 acggcaacat cctggggcac aagctggagt acaacgccat cagcgacaac gtctatatca
     3361 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     3421 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3481 tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg
     3541 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     3601 tggacgagct gtacaagggt accatggcct ccggtaacgc gcgcatcgga aagccagccc
     3661 ctgacttcaa ggccacagcg gtggttgatg gcgccttcaa agaggtgaag ctgtcggact
     3721 acaaagggaa gtacgtggtc ctctttttct accctctgga cttcactttt gtgtgcccca
     3781 ccgagatcat cgcgttcagc aaccgtgcag aggacttccg caagctgggc tgtgaagtgc
     3841 tgggcgtctc ggtggactct cagttcaccc acctggcttg gatcaacacc ccccggaaag
     3901 agggaggctt gggccccctg aacatccccc tgcttgctga cgtgaccaga cgcttgtctg
     3961 aggattacgg cgtgctgaaa acagatgagg gcattgccta caggggcctc tttatcatcg
     4021 atggcaaggg tgtccttcgc cagatcactg ttaatgattt gcctgtggga cgctccgtgg
     4081 atgaggctct gcggctggtc caggccttcc agtacacaga cgagcatggg gaagtttgtc
     4141 ccgctggctg gaagcctggc agtgacacga ttaagcccaa cgtggatgac agcaaggaat
     4201 atttctccaa acacaattaa gcggccgcct cggccaaaca tcgataaaat aaaagatttt
     4261 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     4321 ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag
     4381 atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     4441 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag
     4501 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     4561 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     4621 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     4681 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     4741 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     4801 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     4861 cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca
     4921 ccgacccacc accgggaggt aagctggctg cctcgcgcgt ttcggtgatg acggtgaaaa
     4981 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5041 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     5101 ccagtcacgt agcgatagcg gagtgtactg ataacttcgt ataatgtatg ctatacgaag
     5161 ttattaggtc tgaagaggag tttacgtcca gccaagctag ggccgcgatc cggaaccctt
     5221 aatataactt cgtataatgt atgctatacg aagttatcag tactggctta actatgcggc
     5281 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     5341 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5401 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5461 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5521 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5581 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5641 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5701 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5761 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5821 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5881 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5941 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6001 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6061 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6121 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6181 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6241 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6301 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6361 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6421 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6481 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6541 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6601 gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6661 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6721 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6781 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6841 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6901 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     6961 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7021 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7081 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7141 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7201 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7261 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     7321 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattgct
     7381 agcaattgct agcaattgct agcaattcat accagatcac cgaaaactgt cctccaaatg
     7441 tgtccccctc acactcccaa attcgcgggc ttctgcctct tagaccactc taccctattc
     7501 cccacactca ccggagccaa agccgcggcc cttccgtttc tttgct
//
