LOCUS       sequence-482464-        2814 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cre-dependent expression of a green fluorescent protein biosensor to
            determine mitochondrial cellular reduction-oxidation (redox) 
            states..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2814)
  AUTHORS   Tubert C, Zampese E, Pancani T, Tkatch T, Surmeier DJ
  TITLE     Feed-forward metabotropic signaling by Cav1 Ca(2+) channels supports
            pacemaking in pedunculopontine cholinergic neurons.
  JOURNAL   Neurobiol Dis. 2023 Nov;188:106328. doi: 10.1016/j.nbd.2023.106328. 
            Epub 2023 Oct 16.
  PUBMED    37852390
REFERENCE   2  (bases 1 to 2814)
  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: "Neurobiol 
            Dis."; date: "2023-11"; volume: "188:106328. doi"; pages: " 
            10.1016/j.nbd.2023.106328. Epub 2023 Oct 16"
COMMENT     Sequence Label: pFB-CMV-DIO-MTS-roGFP-WPRE
FEATURES             Location/Qualifiers
     source          1..2814
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        194..497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        498..701
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     652..672
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     698..722
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(803..836)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(887..920)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(929..1642,1643..1645,1646..1648))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(974..995)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1323..1342
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1582..1603
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    1709..1742
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1793..1826
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1851..2439
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1904..1924)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2322..2333)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2465..2484)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2466..2484)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2510..2644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2547..2566)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2601..2620
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   2674..2814
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2674..2803
                     /label=AAV2 ITR
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtagtaat caattacggg gtcattagtt catagcccat
      181 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      241 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      301 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      361 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      421 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      481 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      541 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      601 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      661 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      721 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
      781 gcctccaacc gtcgactccg gaataacttc gtataggata ctttatacga agttatgcag
      841 aatggtagct ggattgtagc tgctattagc aatatgaaac ctcttaataa cttcgtatag
      901 catacattat acgaagttat ggcgcgcctt acttgtacag ctcgtccatg ccgagagtga
      961 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     1021 tgctcagggc ggagcaggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     1081 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt
     1141 ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt
     1201 tgtggcagtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     1261 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     1321 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     1381 gcatggcgga cttgaagaag tcgtgccgct tcatgtggtc ggggtagcgg ctgaagcact
     1441 gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     1501 tgctgatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     1561 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     1621 tgaacagctc ctcgcccttg ctcaccatga tgccgctgcg cttgaagaag cggatgctct
     1681 ggcgcaggct cagcatggtg gcgctagcat aacttcgtat aaagtatcct atacgaagtt
     1741 atttgcctta acccagaaat tatcactgtt attctttaga atggtgcaaa gaataacttc
     1801 gtataatgta tgctatacga agttatgaat tcgatatcaa gcttatcgat aatcaacctc
     1861 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     1921 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     1981 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     2041 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     2101 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     2161 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     2221 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg
     2281 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     2341 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     2401 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgtacccaa
     2461 ttcgccctat agtgagtcgt attacgcgcg cagcggccga ccatggccca acttgtttat
     2521 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2581 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     2641 gatctccgga ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     2701 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     2761 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg cagg
//
