LOCUS       sequence-90322-e        5819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for calcium signaling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5819)
  AUTHORS   Haustein MD, Kracun S, Lu XH, Shih T, Jackson-Weaver O, Tong X, Xu 
            J, Yang XW, O'Dell TJ, Marvin JS, Ellisman MH, Bushong EA, Looger 
            LL, Khakh BS
  TITLE     Conditions and constraints for astrocyte calcium signaling in the 
            hippocampal mossy fiber pathway.
  JOURNAL   Neuron. 2014 Apr 16;82(2):413-29. doi: 10.1016/j.neuron.2014.02.041.
  PUBMED    24742463
REFERENCE   2  (bases 1 to 5819)
  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.neuron.2014.02"; journalName: "Neuron"; date: 
            "2014-04-16- 16"; volume: "82"; issue: "2"; pages: "413-29"
COMMENT     Sequence Label: pZac2.1 gfaABC1D-cyto-GCaMP6f
FEATURES             Location/Qualifiers
     source          1..5819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     intron          1239..1371
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1354..1373
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1416..1434
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1440..1449
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1446..1454,1455..1472,1473..1475,1476..1508,1509..1511,
                     1512..1535,1536..1568,1569..1625,1626..1631,1632..1901,
                     1902..1919,1920..2354,2355..2360,2361..2804)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLASATMVDSSRRKW
                     NKTGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGD
                     GPVLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKG
                     EELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTT
                     LTYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNR
                     IELKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVM
                     RSLGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKD
                     GNGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1548..1557
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1838..1859
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1965..1986)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2226..2245)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2844..2965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2855..2874)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2909..2928
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3127..3256
                     /label=AAV2 ITR
     primer_bind     complement(3275..3292)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3275..3291)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3284..3306)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3433..3888
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3520..3539)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3730..3751
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3914..4018
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4019..4087,4088..4879)
                     /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"
     rep_origin      5050..5638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5539..5558
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatctaa catatcctgg tgtggagtag gggacgctgc
      481 tctgacagag gctcgggggc ctgagctggc tctgtgagct ggggaggagg cagacagcca
      541 ggccttgtct gcaagcagac ctggcagcat tgggctggcc gccccccagg gcctcctctt
      601 catgcccagt gaatgactca ccttggcaca gacacaatgt tcggggtggg cacagtgcct
      661 gcttcccgcc gcaccccagc ccccctcaaa tgccttccga gaagcccatt gagcaggggg
      721 cttgcattgc accccagcct gacagcctgg catcttggga taaaagcagc acagccccct
      781 aggggctgcc cttgctgtgt ggcgccaccg gcggtggaga acaaggctct attcagcctg
      841 tgcccaggaa aggggatcag gggatgccca ggcatggaca gtgggtggca gggggggaga
      901 ggagggctgt ctgcttccca gaagtccaag gacacaaatg ggtgagggga gagctctccc
      961 catagctggg ctgcggccca accccacccc ctcaggctat gccagggggt gttgccaggg
     1021 gcacccgggc atcgccagtc tagcccactc cttcataaag ccctcgcatc ccaggagcga
     1081 gcagagccag agcaggttgg agaggagacg catcacctcc gctgctcgca agctttattg
     1141 cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca acagtctcga
     1201 acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     1261 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     1321 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     1381 cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta taggctagcg
     1441 ccaccatggg ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc
     1501 aaatgggtcg ggatctgtac gacgatgacg ataaggatct cgctagcgcc accatggtcg
     1561 actcatcacg tcgtaagtgg aataagacag gtcacgcagt cagagctata ggtcggctga
     1621 gctcactcga gaacgtctat atcaaggccg acaagcagaa gaacggcatc aaggcgaact
     1681 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgcctaccac taccagcaga
     1741 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcgtgcagt
     1801 ccaaactttc gaaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1861 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggtacc ggagggagca
     1921 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1981 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg
     2041 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2101 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2161 agcacgactt cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct
     2221 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2281 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2341 agctggagta caacctgccg gaccaactga ctgaagagca gatcgcagaa tttaaagagg
     2401 aattctccct atttgacaag gacggggatg ggacaataac aaccaaggag ctggggacgg
     2461 tgatgcggtc tctggggcag aaccccacag aagcagagct gcaggacatg atcaatgaag
     2521 tagatgccga cggtgacggc acaatcgact tccctgagtt cctgacaatg atggcaagaa
     2581 aaatgaaata cagggacacg gaagaagaaa ttagagaagc gttcggtgtg tttgataagg
     2641 atggcaatgg ctacatcagt gcagcagagc ttcgccacgt gatgacaaac cttggagaga
     2701 agttaacaga tgaagaggtt gatgaaatga tcagggaagc agacatcgat ggggatggtc
     2761 aggtaaacta cgaagagttt gtacaaatga tgacagcgaa gtgatctaga gtcgacccgg
     2821 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2881 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2941 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3001 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     3061 cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
     3121 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3181 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3241 gcgagcgagc gcgcagcctt aattaaccta attcactggc cgtcgtttta caacgtcgtg
     3301 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     3361 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     3421 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3481 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3541 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3601 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3661 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3721 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3781 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3841 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     3901 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3961 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4021 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4081 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4141 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4201 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4261 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4321 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4381 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4441 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4501 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4561 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4621 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4681 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4741 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4801 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4861 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4921 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4981 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5041 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5101 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5161 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5221 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5281 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5341 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5401 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5461 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5521 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5581 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5641 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5701 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5761 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
