LOCUS       sequence-500317-        6294 bp    DNA     circular SYN 18-FEB-2026
DEFINITION  mitochondrial calcium sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6294)
  AUTHORS   Kwon SK, Sando R 3rd, Lewis TL, Hirabayashi Y, Maximov A, Polleux F
  TITLE     LKB1 Regulates Mitochondria-Dependent Presynaptic Calcium Clearance 
            and Neurotransmitter Release Properties at Excitatory Synapses along
            Cortical Axons.
  JOURNAL   PLoS Biol. 2016 Jul 18;14(7):e1002516. doi: 
            10.1371/journal.pbio.1002516. eCollection 2016 Jul.
  PUBMED    27429220
REFERENCE   2  (bases 1 to 6294)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 18, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Biol."; date: "2016-07-18"; volume: "14(7):e1002516. doi"; pages: " 
            10.1371/journal.pbio.1002516. eCollection 2016 Jul"
COMMENT     Sequence Label: pCAG mito-GCaMP5G
FEATURES             Location/Qualifiers
     source          1..6294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(42..630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(122..141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(801..1592,1593..1661))
                     /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     1424..1443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1662..1766)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1797..2176
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          2456..3472
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3396..3418
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3480..3499
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3533..3607
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             join(3635..3643,3644..3661,3662..3664,3665..3697,3698..3700,
                     3701..3724,3725..3751,3752..3808,3809..3814,3815..4084,
                     4085..4102,4103..4537,4538..4543,4544..4987)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(3635..3643,3644..3661,3662..3664,3665..3697,3698..3700,
                     3701..3724,3725..3751,3752..3808,3809..3814,3815..4084,
                     4085..4102,4103..4537,4538..4543,4544..4987)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      3731..3740
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4021..4042
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4148..4169)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4409..4428)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(5078..5097)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5143..5198
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5197..5216)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5589..5605)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5589..5605)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5602..5624)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5613..5629
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5637..5643,5644..5661,5662..5667))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5682..5703
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5762..5957
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5808..5943
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5963..6097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6000..6019)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6054..6073
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6165..6182)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
       61 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      481 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      781 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      841 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      901 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      961 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1021 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1081 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1141 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1201 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1261 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1321 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1381 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1441 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1501 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1561 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1621 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1681 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1741 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca
     1801 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1861 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1921 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1981 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2041 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2101 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2161 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     2221 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
     2281 ggcggggggg gggggggggc gcgcgccggg gggggggggg gggggggggg gggggggggg
     2341 ggsgrggsgg rgrggkgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
     2401 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
     2461 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
     2521 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
     2581 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
     2641 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg
     2701 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg
     2761 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg
     2821 tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
     2881 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc
     2941 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc
     3001 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
     3061 gcctcgggcc ggggrgggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
     3121 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
     3181 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
     3241 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
     3301 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
     3361 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
     3421 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     3481 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgctcg agatgtccgt
     3541 cctgacgccg ctgctgctgc ggggcttgac aggctcggcc cggcggctcc cagtgccgcg
     3601 cgccaagatc cattcgttgg gggatccacc ggtcatgggt tctcatcatc atcatcatca
     3661 tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga
     3721 taaggatctc gccaccatgg tcgactcatc acgtcgtaag tggaataaga caggtcacgc
     3781 agtcagagct ataggtcggc tgagctcact cgagaacgtc tatatcaagg ccgacaagca
     3841 gaagaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg gcggcgtgca
     3901 gctcgcctac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3961 caaccactac ctgagcgtgc agtccaaact ttcgaaagac cccaacgaga agcgcgatca
     4021 catggtcctg ctggagttcg tgactgccgc cgggatcact ctcggcatgg acgagctgta
     4081 caagggcggt accggaggga gcatggtgag caagggcgag gagctgttca ccggggtggt
     4141 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     4201 gggtgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     4261 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     4321 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     4381 catccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     4441 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     4501 ggacggcaac atcctggggc acaagctgga gtacaacctg ccggaccaac tgactgaaga
     4561 gcagatcgca gaatttaaag aggctttctc cctatttgac aaggacgggg atgggacaat
     4621 aacaaccaag gagctgggga cggtgatgcg gtctctgggg cagaacccca cagaagcaga
     4681 gctgcaggac atgatcaatg aagtagatgc cgacggtgac ggcacaatcg acttccctga
     4741 gttcctgaca atgatggcaa gaaaaatgaa atacacagac agtgaagaag aaattagaga
     4801 agcgttccgt gtgtttgata aggatggcaa tggctacatc agtgcagcag agcttcgcca
     4861 cgtgatgaca aaccttggag agaagttaac agatgaagag gttgatgaaa tgatcaggga
     4921 agcagacatc gatggggatg gtcaggtaaa ctacgaagag tttgtacaaa tgatgacagc
     4981 gaagtgagcg gccgcaattc actcctcagg tgcaggctgc ctatcagaag gtggtggctg
     5041 gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt
     5101 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     5161 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     5221 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     5281 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     5341 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     5401 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     5461 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     5521 gatccctcga cctgcaccgt cgaccagctg gtcgacggtg caccgtcgac cagcttggcg
     5581 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5641 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5701 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     5761 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5821 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5881 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5941 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     6001 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6061 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     6121 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6181 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6241 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccag
//
