LOCUS       sequence-211046-        5968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Epidermal Mitochondrial-tagged Florescent Calcium Sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5968)
  AUTHORS   Xu S, Chisholm AD
  TITLE     C. elegans epidermal wounding induces a mitochondrial ROS burst that
            promotes wound repair.
  JOURNAL   Dev Cell. 2014 Oct 13;31(1):48-60. doi: 
            10.1016/j.devcel.2014.08.002.
  PUBMED    25313960
REFERENCE   2  (bases 1 to 5968)
  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"; doi: 
            "10.1016/j.devcel.2014.08"; journalName: "Dev Cell"; date: 
            "2014-10-13- 13"; volume: "31"; issue: "1"; pages: "48-60"
COMMENT     Sequence Label: pCZGY2162
FEATURES             Location/Qualifiers
     source          1..5968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             32..106
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             join(131..139,140..157,158..160,161..193,194..196,197..220,
                     221..247,248..304,305..310,311..580,581..598,599..1033,
                     1034..1036,1037..1039,1040..1483)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIMADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSTQCKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(131..139,140..157,158..160,161..193,194..196,197..220,
                     221..247,248..304,305..310,311..580,581..598,599..1033,
                     1034..1036,1037..1039,1040..1483)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIMADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSTQCKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      227..236
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     517..538
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(644..665)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(905..924)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(1499..1523)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1549..1565)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1549..1565)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2302..2322)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2302..2320)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2341..2357)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2341..2357)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2354..2376)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2365..2381
                     /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(2389..2395,2396..2413,2414..2419))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2434..2455
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2572..2589)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2743..3331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2823..2842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3502..4293,4294..4362))
                     /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     4125..4144
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4363..4467)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4493..4948)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4630..4651)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4842..4861
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5075..5097
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5089..5106
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5090..5106
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5116..5135
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5116..5134
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(5961..5968,1..17)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gtacaaaaaa gcaggctccg aattcgccct tatgtccgtc ctgacgccgc tgctgctgcg
       61 gggcttgaca ggctcggccc ggcggctccc agtgccgcgc gccaagatcc attcgttggg
      121 ggatccaatg atgggttctc atcatcatca tcatcatggt atggctagca tgactggtgg
      181 acagcaaatg ggtcgggatc tgtacgacga tgacgataag gatctcgcca ccatggtcga
      241 ctcatcacgt cgtaagtgga ataagacagg tcacgcagtc agagctatag gtcggctgag
      301 ctcactcgag aacgtctata tcatggccga caagcagaag aacggcatca aggcgaactt
      361 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gcctaccact accagcagaa
      421 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtg
      481 caaactttcg aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
      541 cgccgccggg atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat
      601 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      661 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg
      721 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      781 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
      841 gcacgacttc ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt
      901 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      961 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1021 gctggagtac aacacgcgtg accaactgac tgaagagcag atcgcagaat ttaaagaggc
     1081 tttctcccta tttgacaagg acggggatgg gacaataaca accaaggagc tggggacggt
     1141 gatgcggtct ctggggcaga accccacaga agcagagctg caggacatga tcaatgaagt
     1201 agatgccgac ggtaatggca caatcgactt ccctgagttc ctgacaatga tggcaagaaa
     1261 aatgaaagac acagacagtg aagaagaaat tagagaagcg ttccgtgtgt ttgataagga
     1321 tggcaatggc tacatcagtg cagcagagct tcgccacgtg atgacaaacc ttggagagaa
     1381 gttaacagat gaagaggttg atgaaatgat cagggaagca gacatcgatg gggatggtca
     1441 ggtaaactac gaagagtttg tacaaatgat gacagcgaag tgagcaaggg cgaattcgac
     1501 ccagctttct tgtacaaagt ggtgatatcg aattcctgca gcccggggga tccactagtt
     1561 ctagagcggc cgccaccgcg gccgctgtca tcagatcgcc atctcgcgcc cgtgcctctg
     1621 acttctaagt ccaattactc ttcaacatcc ctacatgctc tttctccctg tgctcccacc
     1681 ccctattttt gttattatca aaaaacttct cttaatttct ttgtttttta gcttctttta
     1741 agtcacctct aacaatgaaa ttgtgtagat tcaaaaatag aattaattcg taataaaaag
     1801 tcgaaaaaaa ttgtgctccc tccccccatt aataataatt ctatcccaaa atctacacaa
     1861 tgttctgtgt acacttctta tgttttttac ttctgataaa tttttttgaa acatcataga
     1921 aaaaaccgca cacaaaatac cttatcatat gttacgtttc agtttatgac cgcaattttt
     1981 atttcttcgc acgtctgggc ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt
     2041 ggagtatttt tggaattttt caatcaagtg aaagtttatg aaattaattt tcctgctttt
     2101 gctttttggg gtttccccta ttgtttgtca agatttcgag gacggcgttt ttcttgctaa
     2161 aatcacaagt attgatgagc acgatgcaag aaagatcgga agaaggtttg ggtttgaggc
     2221 tcagtggaag gtgagtagaa gttgataatt tgaaagtgga gtagtgtcta tggggttttt
     2281 gccgagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     2341 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2401 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2461 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2521 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2581 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2641 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2701 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2761 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2821 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2881 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2941 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3001 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3061 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3121 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3181 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3241 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3301 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3361 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3421 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3481 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3541 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3601 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3661 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3721 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3781 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3841 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3901 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3961 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4021 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4081 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4141 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4201 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4261 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4321 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4381 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4441 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     4501 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4561 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4621 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4681 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4741 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4801 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4861 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4921 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
     4981 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     5041 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     5101 gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccag catttgaaaa
     5161 tttgcaccaa tgtattattt aatttttttt ttcgaaaatt taacgcattt tctctctaaa
     5221 aactcgaaat ttagtgtgtt ctaaacaaca gtaagcatac aatacctttg ttcaaaattg
     5281 acgtgctttc tgaaccaata tggttagttt caaaaatttt tgtattatag gatagaaata
     5341 tttggaaata aattttaaaa ccaaacttat gcctttctct tttagtatcc cagctaggta
     5401 atattttagt atttgcccaa atccttgaag taaggagtat ataatttttg aaaaacaata
     5461 aaactccaga taattcatag ttttttctcg aaagaaaatt tttgagatta gttattgaac
     5521 ttcatttttg aacattattc gttgaaaaac actcgctttg tctttatttt caaaaaaatt
     5581 ccgatttccc caaccagaaa aaaaaacaga tagaagaaat ttctccttaa atttcattgt
     5641 ccatctctct tggaaacaca ttatctatca aatgaaaaac gcattttttt tcctggcaga
     5701 aaaatgaaat tggttagatt acactggtta ggtttgaagg tgtaactttc gctttctcag
     5761 caactttcag tataaaagga aacggtcacc atttagaaag acatcagttc atcaactcga
     5821 ggcatgcctg caggtcgact ctagaggatc cccgggattg gccaaaggac ccaaaggtat
     5881 gtttcgaatg atactaacat aacatagaac attttcagga ggacccttgg agggtacagg
     5941 tagaaaaaaa gcttgatatc acaagttt
//
