LOCUS       sequence-202022-        6206 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Presynaptic calcium sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6206)
  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 6206)
  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 vGlut1 GCaMP5G
FEATURES             Location/Qualifiers
     source          1..6206
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      37..46
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1328..1339)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(1738..1746,1747..1764,1765..1767,1768..1800,1801..1803,
                     1804..1827,1828..1854,1855..1911,1912..1917,1918..2187,
                     2188..2205,2206..2640,2641..2646,2647..3090)
                     /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(1738..1746,1747..1764,1765..1767,1768..1800,1801..1803,
                     1804..1827,1828..1854,1855..1911,1912..1917,1918..2187,
                     2188..2205,2206..2640,2641..2646,2647..3090)
                     /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      1834..1843
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2124..2145
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2251..2272)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2512..2531)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(3181..3200)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3246..3301
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3300..3319)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3692..3708)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3692..3708)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3705..3727)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3716..3732
                     /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(3740..3746,3747..3764,3765..3770))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3785..3806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3865..4060
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3911..4046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4066..4200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4103..4122)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4157..4176
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4268..4285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4439..5027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4519..4538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5198..5989,5990..6058))
                     /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     5821..5840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6059..6163)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttggcaaaga attgctcgag ctcaagcttc gaattggcca ccatggagtt ccggcaggag
       61 gagtttcgga agctggcggg gcgcgccctg gggaggctgc accggttact ggagaagcgg
      121 caggaaggcg cggagacatt ggagctgagc gccgacgggc gcccagtgac cacacacacg
      181 cgggacccgc cggtggtgga ctgcacttgc tttggcctcc ctcgccgcta catcatcgcg
      241 atcatgagcg gtctgggttt ctgcatcagc tttggcatcc gctgcaacct gggcgtggcc
      301 atcgtatcca tggtcaacaa cagtacaacc caccgtgggg gccacgtggt ggtgcagaaa
      361 gcccagttca actgggatcc agagactgtc ggcctcatac atggctcctt tttctggggg
      421 tacattgtca ctcagattcc tggaggattt atctgccaaa aattcgcagc caacagggtc
      481 tttggctttg ccattgtggc tacctccacc ctaaatatgt tgatcccttc agcagcccgt
      541 gttcactatg gctgtgtcat cttcgtgagg atccttcagg gattggtgga gggggtcaca
      601 taccctgctt gccatggcat ctggagcaaa tgggcccctc ccttagaacg gagtcggctg
      661 gcgacgacag ccttttgcgg ttcctatgcc ggggcagtgg ttgccatgcc tctggctggg
      721 gtcctggtac agtattcagg atggagttct gtcttctatg tctatggcag cttcgggatc
      781 ttttggtacc tgttctggtt gcttgtctcc tacgagtcac ctgcactaca ccccagcatc
      841 tccgaggagg agcgcaaata cattgaggat gccatcggag aaagcgccaa gctcatgaac
      901 cctgttacga agtttaacac accctggagg cgcttcttta cctccatgcc ggtctatgcc
      961 atcattgtcg ccaacttttg ccgcagctgg actttctacc tgctcctcat ctcccagccc
     1021 gcctactttg aagaagtgtt cggctttgag atcagcaagg tgggactggt gtcggcactg
     1081 cctcaccttg tcatgactat catcgtaccc atcggaggcc agatcgccga cttcctgcgc
     1141 agtcgtcata taatgtccac gaccaatgtg cgaaagctga tgaactgcgg gggtttcggg
     1201 atggaagcta cgctgctgct ggtggtcgga tactcacact ccaagggcgt ggccatctcc
     1261 ttcctggtcc tggctgtggg cttcagtggc tttgctatct ctgggtttaa cgtgaaccac
     1321 ttggacatcg cccctcgata tgccagcatc ttgatgggca tttccaatgg cgtgggcaca
     1381 ctgtctggga tggtgtgccc catcatcgtg ggtgcaatga ccaagcacaa gacgcgggag
     1441 gagtggcagt acgtgttcct catagcctcc ctggtgcact atggaggtgt catcttctat
     1501 ggggtctttg cttcgggaga gaaacagccg tgggcagagc cggaggagat gagcgaggag
     1561 aagtgtggct ttgttggcca cgaccagctg gctggcagtg acgaaagtga aatggaagac
     1621 gaggttgagc ccccgggggc accccccgca cctccgcctt cctacggggc cacacacagc
     1681 acagttcagc ctccaaggcc cccaccccct gtccgggact acgaattcgg tggttctatg
     1741 ggttctcatc atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt
     1801 cgggatctgt acgacgatga cgataaggat ctcgccacca tggtcgactc atcacgtcgt
     1861 aagtggaata agacaggtca cgcagtcaga gctataggtc ggctgagctc actcgagaac
     1921 gtctatatca aggccgacaa gcagaagaac ggcatcaagg cgaacttcaa gatccgccac
     1981 aacatcgagg acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc
     2041 gacggccccg tgctgctgcc cgacaaccac tacctgagcg tgcagtccaa actttcgaaa
     2101 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2161 actctcggca tggacgagct gtacaagggc ggtaccggag ggagcatggt gagcaagggc
     2221 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2281 cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca cctacggcaa gctgaccctg
     2341 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2401 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2461 aagtccgcca tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc
     2521 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2581 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2641 ctgccggacc aactgactga agagcagatc gcagaattta aagaggcttt ctccctattt
     2701 gacaaggacg gggatgggac aataacaacc aaggagctgg ggacggtgat gcggtctctg
     2761 gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga tgccgacggt
     2821 gacggcacaa tcgacttccc tgagttcctg acaatgatgg caagaaaaat gaaatacaca
     2881 gacagtgaag aagaaattag agaagcgttc cgtgtgtttg ataaggatgg caatggctac
     2941 atcagtgcag cagagcttcg ccacgtgatg acaaaccttg gagagaagtt aacagatgaa
     3001 gaggttgatg aaatgatcag ggaagcagac atcgatgggg atggtcaggt aaactacgaa
     3061 gagtttgtac aaatgatgac agcgaagtga gcggccgcaa ttcactcctc aggtgcaggc
     3121 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     3181 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     3241 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     3301 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     3361 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     3421 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     3481 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     3541 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     3601 agctgtccct cttctcttat ggagatccct cgacctgcac cgtcgaccag ctggtcgacg
     3661 gtgcaccgtc gaccagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3721 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3781 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3841 gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct
     3901 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3961 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4021 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca
     4081 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4141 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     4201 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4261 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4321 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4381 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4441 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4501 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4561 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4621 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4681 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4741 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4801 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4861 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4921 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4981 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5041 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5101 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5161 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5221 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5281 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5341 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5401 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5461 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5521 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5581 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5641 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5701 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5761 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5821 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5881 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5941 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6001 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6061 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6121 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6181 tgccacctgg gtcgacattg attatt
//
