LOCUS       sequence-244048-        6490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Highly resposive GFP-based pyruvate nanosensor with a nuclear 
            localization signal..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6490)
  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 6490)
  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: NLS-PyronicSF/pCMV-myc-nuc
FEATURES             Location/Qualifiers
     source          1..6490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      650..659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2201..2221
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2225..2245
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2249..2269
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2282..2311
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2347..2364)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2353..2577
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2623..3051
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2710..2729)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2920..2941
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3060..3080)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3065..3394
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3245..3380
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3307..3326
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3420..4214
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3474..3493)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4084..4103
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4388..4509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4425..4444)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4479..4498
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4542..4559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4713..5301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4793..4812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5472..6263,6264..6332))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6095..6114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6333..6437)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtaccgaatt cacattgatt attgagtagt tattaatagt aatcaattac ggggtcatta
       61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      361 atctacggtt agtcatcgct attaccatag tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctttctggc
      601 taactagaga acccactgct tactggcacg tggaaattaa tacgacgtgg ccaccatggc
      661 ccaggtgcag ctgcaggtcg acctcgagac catggcatat agcaaaattc ggcagcccaa
      721 actgagcgat gtgattgagc agcagctgga gtttctgatt ctggaaggca ccctgagacc
      781 tggagagaaa ctgccccctg agcgcgaact cgccaagcag ttcgacgtga gtcgaccatc
      841 actgagggag gctatccaga ggctggaagc aaagggactg ctcctgagga gacagggagg
      901 agggactttc gtgcagagct ccctgtggca gagcttcagc gaccccctgg tcgagctcct
      961 gtctgaccac ccagaaagtc agtacgatct cctggagaca agacatgctc tggaaggcat
     1021 cgccgcttac tatgcagccc tgcggtccac tgacgaggat aaggaacgca tccgagagct
     1081 gcaccatgcc attgaactcg ctcagcagtc aggagacctg gatgcagaga gcaacgccgt
     1141 gctgcagtac cagattgcag tcaccgaggc tgcacacaat gtggtcctcc tgcatctcct
     1201 gaggtgcatg gagccaatgc tggcccagaa cgtgagacag aattttgagc tcctgctcga
     1261 aaacgtctat atcaaggccg acaagcagaa aaacggcatt aaggctaact tcaagatcag
     1321 acacaacatc gaggatggtg gcgtgcagct ggcctaccat tatcagcaga acacaccaat
     1381 cggagacgga ccagtgctgc tcccagataa tcactacctg agcgtccagt ccaagctgtc
     1441 taaagaccct aacgagaagc gggatcatat ggtgctgctc gaatttgtca cagccgctgg
     1501 gatcactctg ggtatggacg agctctataa aggagggacc ggtggcagta tggtgtcaaa
     1561 gggcgaggaa ctgttcacag gagtggtccc cattctggtg gagctcgacg gcgatgtcaa
     1621 tggacacaaa ttttccgtgt ctggcgaggg cgaaggagat gctacctacg ggaagctgac
     1681 actcaaattc atctgcacca caggcaagct gccagtgccc tggcctactc tggtcactac
     1741 cctcacctac ggggtgcagt gtttctccag atatcccgac cacatgaagc agcatgattt
     1801 ctttaaatct gctatgcctg aggggtacat ccaggaacgg acaattttct ttaaggacga
     1861 tggtaactac aaaacacgcg cagaggtgaa gttcgaaggc gacactctgg tcaatcgaat
     1921 cgagctgaag ggaattgact ttaaagaaga tgggaacatc ctgggtcaca agctggagta
     1981 caatactagg tattctcggc gcgaaatgct gccactcgtg tctagtcaca ggaccagaat
     2041 ctttgaggca attatggccg gaaagcccga ggaagctaga gaagcaagtc accggcatct
     2101 ggccttcatc gaggaaattc tgctcgaccg gagccgcgag gaatcccgaa gggagcgcag
     2161 cctgagacga ctcgaacagc gaaagaacgc ggccgcagat ccaaaaaaga agagaaaggt
     2221 agatccaaaa aagaagagaa aggtagatcc aaaaaagaag agaaaggtag atacggccgc
     2281 agaacaaaaa ctcatctcag aagaggatct gaatggggcc gcatagtcta gaagctcgct
     2341 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2401 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2461 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2521 agggggagga ttgggaagac aatagcaggc atgctgggga tggcccgggc tctatggctt
     2581 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2641 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2701 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2761 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctct
     2821 cccccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2881 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2941 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3001 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3061 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3121 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3181 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3241 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgcccctag gctgactaat
     3301 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3361 aggaggcttt tttggaggcc taggcttttg caaaaagctc cccccccggg aggtccacaa
     3421 tggttgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3481 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3541 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactcc
     3601 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3661 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3721 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3781 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3841 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3901 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgt atgcccgacg
     3961 gcgaggatct cgtcgtgact catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4021 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4081 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4141 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4201 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4261 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4321 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4381 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4441 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4501 tgtatcttat catgtctgta taccgtcgat ctttccgctt cctcgctcac tgactcgctg
     4561 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4621 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4681 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4741 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4801 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4861 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4921 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtatgca cgaacccccc
     4981 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5041 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5101 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5161 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5221 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5281 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5341 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5401 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5461 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5521 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5581 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5641 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5701 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5761 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5821 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5881 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5941 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6001 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6061 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6121 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6181 ctgttgagat ccagttcgat gtaacccact cgggcaccca actgatcttc agcatctttt
     6241 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6301 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6361 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6421 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcag atcgacggat
     6481 cgggagatcg
//
