LOCUS       sequence-262053-        6648 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 6648)
  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 6648)
  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-PAmCherry
FEATURES             Location/Qualifiers
     source          1..6648
                     /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"
     promoter        528..806
                     /label=chicken beta-actin promoter
     primer_bind     1062..1081
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1136..1145
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1157..1213
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     1426..1447
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1553..1574)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1814..1833)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2405..2461
                     /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"
     regulatory      2468..2477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2474..3184
                     /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(2619..2637)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2887..2906
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3210..3331
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3221..3240)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3275..3294
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3417..3434)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3423..3630
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3660..3800
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3660..3789
                     /label=AAV2 ITR
     rep_origin      3886..4341
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3973..3992)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4183..4204
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4408..4427)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4527..4549
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4587..4605)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4673..4777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4778..4846,4847..5638)
                     /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(4996..5015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5809..6397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6298..6317
                     /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 gggggggggg cgcgcgccag
      661 gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca
      721 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct
      781 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc
      841 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
      901 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc
      961 ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagcta gagcctctgc
     1021 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt
     1081 gctgtctcat cattttggca aagaattcct cgaagatccg aaggggttca agcttgccac
     1141 catggtcgac tcatcacgtc gtaagtggaa taagacaggt cacgcagtca gagctatagg
     1201 tcggctgagc tcactcgaga acgtctatat caaggccgac aagcagaaga acggcatcaa
     1261 ggcgaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg cctaccacta
     1321 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1381 cgtgcagtcc aaactttcga aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1441 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg
     1501 agggagcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1561 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg agggcgatgc
     1621 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1681 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1741 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacatcc aggagcgcac
     1801 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1861 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1921 ggggcacaag ctggagtaca acctgccgga ccaactgact gaagagcaga tcgcagaatt
     1981 taaagaggaa ttctccctat ttgacaagga cggggatggg acaataacaa ccaaggagct
     2041 ggggacggtg atgcggtctc tggggcagaa ccccacagaa gcagagctgc aggacatgat
     2101 caatgaagta gatgccgacg gtgacggcac aatcgacttc cctgagttcc tgacaatgat
     2161 ggcaagaaaa atgaaataca gggacacgga agaagaaatt agagaagcgt tcggtgtgtt
     2221 tgataaggat ggcaatggct acatcagtgc agcagagctt cgccacgtga tgacaaacct
     2281 tggagagaag ttaacagatg aagaggttga tgaaatgatc agggaagcag acatcgatgg
     2341 ggatggtcag gtaaactacg aagagtttgt acaaatgatg acagcgaagg aattcggaag
     2401 cggagctact aacttcagcc tgctgaagca ggctggagac gtggaggaga accctggacc
     2461 tggatccgcc accatggtga gcaagggcga ggaggataac atggccatca ttaaggagtt
     2521 catgcgcttc aaggtgcaca tggaggggtc cgtgaacggc cacgtgttcg agatcgaggg
     2581 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     2641 tggccccctg cccttcacct gggacatcct gtcccctcaa ttcatgtacg gctccaatgc
     2701 ctacgtgaag caccccgccg acatccccga ctactttaag ctgtccttcc ccgagggctt
     2761 caagtgggag cgcgtgatga aattcgagga cggcggcgtg gtgaccgtga cccaggactc
     2821 ctccctgcag gacggtgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     2881 cgacggcccc gtaatgcaga agaagaccat gggctgggag gccctctccg agcggatgta
     2941 ccccgaggac ggcgccctga agggcgaggt caagccgaga gtgaagctga aggacggcgg
     3001 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     3061 cgcctacaac gtcaaccgca agttggacat cacctcacac aacgaggact acaccatcgt
     3121 ggaacagtac gaacgtgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     3181 gtaagcggcc gcgcggatcc cagacatgat aagatacatt gatgagtttg gacaaaccac
     3241 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3301 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3361 tcaggttcag ggggaggtgt gggaggtttt ttagtcgact agagctcgct gatcagcctc
     3421 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3481 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3541 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3601 ttgggaagac aatagcaggc atgctgggga gagatctcac gtgcggaccg agcggccgca
     3661 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     3721 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     3781 agcgcgcagc tgcctgcagg ggcgcctgat gccagctggc gtaatagcga agaggcccgc
     3841 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg
     3901 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3961 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4021 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4081 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4141 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4201 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4261 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4321 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4381 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4441 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4501 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4561 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4621 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4681 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4741 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4801 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4861 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4921 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4981 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5041 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5101 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5161 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5221 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5281 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5341 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5401 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5461 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5521 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5581 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5641 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5701 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5761 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5821 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5881 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5941 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6001 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6061 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6121 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6181 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6241 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6301 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6361 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6421 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6481 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6541 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     6601 tctccccgcg cgttggccga ttcattaatg cagctggggc tcacatgt
//
