LOCUS       sequence-199363-        6289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mitochondrial calcium sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6289)
  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 6289)
  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: "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..6289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(42..629)
                     /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(800..1591,1592..1660))
                     /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     1423..1442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1661..1765)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1796..2175
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2177..2454
                     /label=chicken beta-actin promoter
     intron          2455..3467
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3391..3413
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3475..3494
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3528..3602
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             join(3630..3638,3639..3656,3657..3659,3660..3692,3693..3695,
                     3696..3719,3720..3746,3747..3803,3804..3809,3810..4079,
                     4080..4097,4098..4532,4533..4538,4539..4982)
                     /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(3630..3638,3639..3656,3657..3659,3660..3692,3693..3695,
                     3696..3719,3720..3746,3747..3803,3804..3809,3810..4079,
                     4080..4097,4098..4532,4533..4538,4539..4982)
                     /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      3726..3735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4016..4037
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4143..4164)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4404..4423)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(5073..5092)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5138..5193
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5192..5211)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5584..5600)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5584..5600)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5597..5619)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5608..5624
                     /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(5632..5638,5639..5656,5657..5662))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5677..5698
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5757..5952
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5803..5938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5958..6092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5995..6014)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6049..6068
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6160..6177)
                     /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 agacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
      541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
      601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
      661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
      721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
      781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
      841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
      901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
      961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     1681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     1741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgacat
     1801 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     1861 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     1921 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     1981 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     2041 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     2101 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     2161 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
     2221 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
     2281 gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
     2341 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
     2401 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     2461 gctgcgttgc cttcgccccg tgccccgctc cgcgccgcct cgcgccgccc gccccggctc
     2521 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     2581 aattagcgct tggtttaatg acggctcgtt tcttttctgt ggctgcgtga aagccttaaa
     2641 gggctccggg agggcccttt gtgcgggggg gagcggctcg gggggtgcgt gcgtgtgtgt
     2701 gtgcgtgggg agcgccgcgt gcggcccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     2761 ggcgcggggc tttgtgcgct ccgcgtgtgc gcgaggggag cgcggccggg ggcggtgccc
     2821 cgcggtgcgg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg
     2881 gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac ccccctcccc
     2941 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     3001 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
     3061 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
     3121 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     3181 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
     3241 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
     3301 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     3361 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     3421 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     3481 tgctggttat tgtgctgtct catcattttg gcaaagaatt gctcgagatg tccgtcctga
     3541 cgccgctgct gctgcggggc ttgacaggct cggcccggcg gctcccagtg ccgcgcgcca
     3601 agatccattc gttgggggat ccaccggtca tgggttctca tcatcatcat catcatggta
     3661 tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg
     3721 atctcgccac catggtcgac tcatcacgtc gtaagtggaa taagacaggt cacgcagtca
     3781 gagctatagg tcggctgagc tcactcgaga acgtctatat caaggccgac aagcagaaga
     3841 acggcatcaa ggcgaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     3901 cctaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3961 actacctgag cgtgcagtcc aaactttcga aagaccccaa cgagaagcgc gatcacatgg
     4021 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     4081 gcggtaccgg agggagcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     4141 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg
     4201 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     4261 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     4321 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacatcc
     4381 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     4441 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     4501 gcaacatcct ggggcacaag ctggagtaca acctgccgga ccaactgact gaagagcaga
     4561 tcgcagaatt taaagaggct ttctccctat ttgacaagga cggggatggg acaataacaa
     4621 ccaaggagct ggggacggtg atgcggtctc tggggcagaa ccccacagaa gcagagctgc
     4681 aggacatgat caatgaagta gatgccgacg gtgacggcac aatcgacttc cctgagttcc
     4741 tgacaatgat ggcaagaaaa atgaaataca cagacagtga agaagaaatt agagaagcgt
     4801 tccgtgtgtt tgataaggat ggcaatggct acatcagtgc agcagagctt cgccacgtga
     4861 tgacaaacct tggagagaag ttaacagatg aagaggttga tgaaatgatc agggaagcag
     4921 acatcgatgg ggatggtcag gtaaactacg aagagtttgt acaaatgatg acagcgaagt
     4981 gagcggccgc aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     5041 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     5101 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     5161 gcaatagtgt gttggaattt tttgtgtcta tcactcggaa ggacatatgg gagggcaaat
     5221 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     5281 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     5341 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     5401 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     5461 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     5521 ctcgacctgc accgtcgacc agctggtcga cggtgcaccg tcgaccagct tggcgtaatc
     5581 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5641 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5701 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat
     5761 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5821 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5881 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5941 ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6001 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6061 aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc
     6121 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6181 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccag
//
