LOCUS       sequence-199086-        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"
     repeat_region   32..161
                     /label=AAV2 ITR
     intron          1012..1144
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1127..1146
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1189..1207
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1213..1222
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1219..1227,1228..1245,1246..1248,1249..1281,1282..1284,
                     1285..1308,1309..1341,1342..1398,1399..1404,1405..1674,
                     1675..1692,1693..2127,2128..2133,2134..2577)
                     /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      1321..1330
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1611..1632
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1738..1759)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1999..2018)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2617..2738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2628..2647)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2682..2701
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2900..3029
                     /label=AAV2 ITR
     primer_bind     complement(3048..3065)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3048..3064)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3057..3079)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3206..3661
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3293..3312)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3503..3524
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3687..3791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3792..3860,3861..4652)
                     /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      4823..5411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5312..5331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5699..5720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5735..5740,5741..5758,5759..5765)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5773..5789
                     /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     5778..5800
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5797..5813
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5797..5813
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tacgccagat ttaattaagg ccttaattag gctgcgcgct cgctcgctca ctgaggccgc
       61 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
      121 gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa tgattaaccc
      181 gccatgctac ttatctacgt agccatgctc taggaagatc taacatatcc tggtgtggag
      241 taggggacgc tgctctgaca gaggctcggg ggcctgagct ggctctgtga gctggggagg
      301 aggcagacag ccaggccttg tctgcaagca gacctggcag cattgggctg gccgcccccc
      361 agggcctcct cttcatgccc agtgaatgac tcaccttggc acagacacaa tgttcggggt
      421 gggcacagtg cctgcttccc gccgcacccc agcccccctc aaatgccttc cgagaagccc
      481 attgagcagg gggcttgcat tgcaccccag cctgacagcc tggcatcttg ggataaaagc
      541 agcacagccc cctaggggct gcccttgctg tgtggcgcca ccggcggtgg agaacaaggc
      601 tctattcagc ctgtgcccag gaaaggggat caggggatgc ccaggcatgg acagtgggtg
      661 gcaggggggg agaggagggc tgtctgcttc ccagaagtcc aaggacacaa atgggtgagg
      721 ggagagctct ccccatagct gggctgcggc ccaaccccac cccctcaggc tatgccaggg
      781 ggtgttgcca ggggcacccg ggcatcgcca gtctagccca ctccttcata aagccctcgc
      841 atcccaggag cgagcagagc cagagcaggt tggagaggag acgcatcacc tccgctgctc
      901 gcaagcttta ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac
      961 acaacagtct cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc
     1021 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
     1081 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
     1141 acaggtgtcc actcccagtt caattacagc tcttaaggct agagtactta atacgactca
     1201 ctataggcta gcgccaccat gggttctcat catcatcatc atcatggtat ggctagcatg
     1261 actggtggac agcaaatggg tcgggatctg tacgacgatg acgataagga tctcgctagc
     1321 gccaccatgg tcgactcatc acgtcgtaag tggaataaga caggtcacgc agtcagagct
     1381 ataggtcggc tgagctcact cgagaacgtc tatatcaagg ccgacaagca gaagaacggc
     1441 atcaaggcga acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgcctac
     1501 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1561 ctgagcgtgc agtccaaact ttcgaaagac cccaacgaga agcgcgatca catggtcctg
     1621 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt
     1681 accggaggga gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1741 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggtgagggc
     1801 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1861 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     1921 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta catccaggag
     1981 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2041 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2101 atcctggggc acaagctgga gtacaacctg ccggaccaac tgactgaaga gcagatcgca
     2161 gaatttaaag aggaattctc cctatttgac aaggacgggg atgggacaat aacaaccaag
     2221 gagctgggga cggtgatgcg gtctctgggg cagaacccca cagaagcaga gctgcaggac
     2281 atgatcaatg aagtagatgc cgacggtgac ggcacaatcg acttccctga gttcctgaca
     2341 atgatggcaa gaaaaatgaa atacagggac acggaagaag aaattagaga agcgttcggt
     2401 gtgtttgata aggatggcaa tggctacatc agtgcagcag agcttcgcca cgtgatgaca
     2461 aaccttggag agaagttaac agatgaagag gttgatgaaa tgatcaggga agcagacatc
     2521 gatggggatg gtcaggtaaa ctacgaagag tttgtacaaa tgatgacagc gaagtgatct
     2581 agagtcgacc cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     2641 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     2701 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     2761 ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca
     2821 aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc
     2881 gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     2941 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3001 gcggcctcag tgagcgagcg agcgcgcagc cttaattaac ctaattcact ggccgtcgtt
     3061 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     3121 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3181 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3241 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3301 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3361 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3421 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3481 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3541 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3601 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3661 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3721 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3781 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3841 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3901 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3961 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4021 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4081 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4141 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4201 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4261 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4321 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4381 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4441 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4501 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4561 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4621 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4681 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4741 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5461 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5521 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5581 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5641 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5701 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5761 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgat
//
