LOCUS       sequence-244362-        5311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Highly resposive GFP-based pyruvate nanosensor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5311)
  AUTHORS   Arce-Molina R, Cortes-Molina F, Sandoval PY, Galaz A, Alegria K, 
            Schirmeier S, Barros LF, San Martin A
  TITLE     A highly responsive pyruvate sensor reveals pathway-regulatory role 
            of the mitochondrial pyruvate carrier MPC.
  JOURNAL   eLife. 2020 Mar 6;9. pii: 53917. doi: 10.7554/eLife.53917.
  PUBMED    32142409
REFERENCE   2  (bases 1 to 5311)
  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: "eLife."; 
            date: "2020-03-6"; volume: "9. pii: 53917. doi"; pages: " 
            10.7554/eLife.53917"
COMMENT     Sequence Label: PyronicSF-mRuby2/pBI-CMV1
FEATURES             Location/Qualifiers
     source          1..5311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2251..2332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2343..2360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2514..3102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2594..2613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3273..4064,4065..4133))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3896..3915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4134..4238)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    4271..4352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4475..5185)
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
ORIGIN
        1 ctcgagtagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gggatccctc gagatggcat atagcaaaat tcggcagccc aaactgagcg atgtgattga
      661 gcagcagctg gagtttctga ttctggaagg caccctgaga cctggagaga aactgccccc
      721 tgagcgcgaa ctcgccaagc agttcgacgt gagtcgacca tcactgaggg aggctatcca
      781 gaggctggaa gcaaagggac tgctcctgag gagacaggga ggagggactt tcgtgcagag
      841 ctccctgtgg cagagcttca gcgaccccct ggtcgagctc ctgtctgacc acccagaaag
      901 tcagtacgat ctcctggaga caagacatgc tctggaaggc atcgccgctt actatgcagc
      961 cctgcggtcc actgacgagg ataaggaacg catccgagag ctgcaccatg ccattgaact
     1021 cgctcagcag tcaggagacc tggatgcaga gagcaacgcc gtgctgcagt accagattgc
     1081 agtcaccgag gctgcacaca atgtggtcct cctgcatctc ctgaggtgca tggagccaat
     1141 gctggcccag aacgtgagac agaattttga gctcctgctc gaaaacgtct atatcaaggc
     1201 cgacaagcag aaaaacggca ttaaggctaa cttcaagatc agacacaaca tcgaggatgg
     1261 tggcgtgcag ctggcctacc attatcagca gaacacacca atcggagacg gaccagtgct
     1321 gctcccagat aatcactacc tgagcgtcca gtccaagctg tctaaagacc ctaacgagaa
     1381 gcgggatcat atggtgctgc tcgaatttgt cacagccgct gggatcactc tgggtatgga
     1441 cgagctctat aaaggaggga ccggtggcag tatggtgtca aagggcgagg aactgttcac
     1501 aggagtggtc cccattctgg tggagctcga cggcgatgtc aatggacaca aattttccgt
     1561 gtctggcgag ggcgaaggag atgctaccta cgggaagctg acactcaaat tcatctgcac
     1621 cacaggcaag ctgccagtgc cctggcctac tctggtcact accctcacct acggggtgca
     1681 gtgtttctcc agatatcccg accacatgaa gcagcatgat ttctttaaat ctgctatgcc
     1741 tgaggggtac atccaggaac ggacaatttt ctttaaggac gatggtaact acaaaacacg
     1801 cgcagaggtg aagttcgaag gcgacactct ggtcaatcga atcgagctga agggaattga
     1861 ctttaaagaa gatgggaaca tcctgggtca caagctggag tacaatacta ggtattctcg
     1921 gcgcgaaatg ctgccactcg tgtctagtca caggaccaga atctttgagg caattatggc
     1981 cggaaagccc gaggaagcta gagaagcaag tcaccggcat ctggccttca tcgaggaaat
     2041 tctgctcgac cggagccgcg aggaatcccg aagggagcgc agcctgagac gactcgaaca
     2101 gcgaaagaac tgactcgaga agcttgtcga cgatatctcc agaggatcat aatcagccat
     2161 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     2221 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     2281 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gccccgagct
     2341 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2401 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2461 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2521 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2581 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2641 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2701 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2761 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2821 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2881 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2941 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3001 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3061 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3121 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3181 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3241 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3301 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3361 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3421 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3481 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3541 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3601 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3661 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3721 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3781 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3841 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     3901 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3961 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4021 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4081 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4141 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4201 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4261 cctgacgtcg gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     4321 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     4381 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     4441 gtatggctga ttatgatcct ctagactgca gttatttata cagttcatcc atcccgcctc
     4501 ccagtcctgc gaattttgcg acagcgtgct cccgctgcac cacgaacatc tcgttatcgg
     4561 actcctccag tctctccagc ctgtggtcca ctgcgtggat gcctggcatc ttgatattgc
     4621 ccactgtctt cttgctccgg tatgtggtca caaagctaca agacaggtgt cctcctccat
     4681 ccaccttcag ggccatgtgg gtatagcctc tcaggccgcc gtcggctggg tacatcatct
     4741 ctgtgttagg ctcccagccc ttggtcttct tctgcatcac ggggccatta gatgggaagt
     4801 tcacgcctct cacctgcacg tgatacacca ggcagccatc ctccagggat gtgtcctgca
     4861 tcacggtcac cacgccgcca tcctcgtagc gtgtcacccg ctcccaggta aagccctctg
     4921 ggaagctctg cttaaagaag tcagggatgc ccttgggata cttgatgaag gtcctgctgc
     4981 cgtacatgaa agaggtggcc aggatgtcaa aggcgaaggg cagtggtcct ccctcgatca
     5041 ctttgatcct cattgtctgg gtgcccatgt aaggattgcc ctcgccctcg ccggtacact
     5101 taaactggtg gccgttcacg ctgccctcca tgaccacctt catgcgcata ttctccttga
     5161 tcagctcctc gcccttggac accatggtct gcagcctcag gagatctggg cccccgcggc
     5221 atatgaccgg tgaattctcc aggcgatctg acggttcact aaacgagctc tgcttatata
     5281 ggcctcccac cgtacacgcc acctcgacat a
//
