LOCUS       sequence-261974-        7062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for calcium imaging and photoactivation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7062)
  AUTHORS   Lee D, Kume M, Holy TE
  TITLE     Sensory coding mechanisms revealed by optical tagging of 
            physiologically defined neuronal types.
  JOURNAL   Science. 2019 Dec 13;366(6471):1384-1389. doi: 
            10.1126/science.aax8055.
  PUBMED    31831669
REFERENCE   2  (bases 1 to 7062)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2019-12-13- 13"; volume: "366"; 
            issue: "6471"; pages: "1384-1389"
COMMENT     Sequence Label: pAAV2-GCaMP6f-P2A-H2B-PAmCherry
FEATURES             Location/Qualifiers
     source          1..7062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        223..526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1089..1108
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1163..1172
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1184..1240
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     1453..1474
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1580..1601)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1841..1860)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2432..2488
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2501..2878
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      2891..2900
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2897..3607
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein"
                     /label=PAmCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
                     GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3042..3060)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3310..3329
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3624..3745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3635..3654)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3689..3708
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3831..3848)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3837..4044
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4074..4214
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4074..4203
                     /label=AAV2 ITR
     rep_origin      4300..4755
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4387..4406)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4597..4618
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4822..4841)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4941..4963
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5001..5019)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5087..5191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5192..5260,5261..6052)
                     /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     complement(5410..5429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6223..6811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6712..6731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtagatct gaattcggta cctagttatt aatagtaatc
      181 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      241 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      301 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      361 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      421 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      481 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      541 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      601 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggy gggsssnsgg
      661 nngggggggg gggggggcgs gcgccaggcg gggcggggcg gggcgagggg cggggcgggg
      721 cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta
      781 tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg
      841 ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
      901 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
      961 gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg
     1021 aggggctccg ggagctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     1081 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcctcga
     1141 agatccgaag gggttcaagc ttgccaccat ggtcgactca tcacgtcgta agtggaataa
     1201 gacaggtcac gcagtcagag ctataggtcg gctgagctca ctcgagaacg tctatatcaa
     1261 ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga
     1321 cggcggcgtg cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt
     1381 gctgctgccc gacaaccact acctgagcgt gcagtccaaa ctttcgaaag accccaacga
     1441 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1501 ggacgagctg tacaagggcg gtaccggagg gagcatggtg agcaagggcg aggagctgtt
     1561 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1621 cgtgtccggc gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1681 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1741 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1801 gcccgaaggc tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1861 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1921 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaacc tgccggacca
     1981 actgactgaa gagcagatcg cagaatttaa agaggaattc tccctatttg acaaggacgg
     2041 ggatgggaca ataacaacca aggagctggg gacggtgatg cggtctctgg ggcagaaccc
     2101 cacagaagca gagctgcagg acatgatcaa tgaagtagat gccgacggtg acggcacaat
     2161 cgacttccct gagttcctga caatgatggc aagaaaaatg aaatacaggg acacggaaga
     2221 agaaattaga gaagcgttcg gtgtgtttga taaggatggc aatggctaca tcagtgcagc
     2281 agagcttcgc cacgtgatga caaaccttgg agagaagtta acagatgaag aggttgatga
     2341 aatgatcagg gaagcagaca tcgatgggga tggtcaggta aactacgaag agtttgtaca
     2401 aatgatgaca gcgaaggaat tcggaagcgg agctactaac ttcagcctgc tgaagcaggc
     2461 tggagacgtg gaggagaacc ctggacctga tatcgccacc atgccagagc cagcgaagtc
     2521 tgctcccgcc ccgaaaaagg gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg
     2581 caagaagcgc aagcgcagcc gcaaggagag ctattccatc tatgtgtaca aggttctgaa
     2641 gcaggtccac cctgacaccg gcatttcgtc caaggccatg ggcatcatga attcgtttgt
     2701 gaacgacatt ttcgagcgca tcgcaggtga ggcttcccgc ctggcgcatt acaacaagcg
     2761 ctcgaccatc acctccaggg agatccagac ggccgtgcgc ctgctgctgc ctggggagtt
     2821 ggccaagcac gccgtgtccg agggtactaa ggccatcacc aagtacacca gcgctaagga
     2881 tccaccggtc gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcattaagga
     2941 gttcatgcgc ttcaaggtgc acatggaggg gtccgtgaac ggccacgtgt tcgagatcga
     3001 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     3061 gggtggcccc ctgcccttca cctgggacat cctgtcccct caattcatgt acggctccaa
     3121 tgcctacgtg aagcaccccg ccgacatccc cgactacttt aagctgtcct tccccgaggg
     3181 cttcaagtgg gagcgcgtga tgaaattcga ggacggcggc gtggtgaccg tgacccagga
     3241 ctcctccctg caggacggtg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     3301 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggccctct ccgagcggat
     3361 gtaccccgag gacggcgccc tgaagggcga ggtcaagccg agagtgaagc tgaaggacgg
     3421 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     3481 cggcgcctac aacgtcaacc gcaagttgga catcacctca cacaacgagg actacaccat
     3541 cgtggaacag tacgaacgtg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     3601 caagtaaact agtccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3661 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3721 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3781 tcagggggag gtgtgggagg ttttttagtc gactagagct cgctgatcag cctcgactgt
     3841 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3901 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3961 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4021 agacaatagc aggcatgctg gggagagatc tcacgtgcgg accgagcggc cgcaggaacc
     4081 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4141 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4201 cagctgcctg caggggcgcc tgatgccagc tggcgtaata gcgaagaggc ccgcaccgat
     4261 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat
     4321 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4381 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4441 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4501 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4561 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4621 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4681 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4741 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4801 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4861 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4921 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4981 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5041 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5101 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5161 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5221 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5281 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5341 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5401 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5461 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5521 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5581 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5641 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5701 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5761 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5821 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5881 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5941 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6001 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6061 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6121 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6181 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6241 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6301 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6361 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6841 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6901 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6961 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     7021 cgcgcgttgg ccgattcatt aatgcagctg gggctcacat gt
//
