LOCUS       sequence-325118-        7580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the endocannabinoid sensor GRAB_eCB2.0 in CaMKII positive 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7580)
  AUTHORS   Dong A, He K, Dudok B, Farrell JS, Guan W, Liput DJ, Puhl HL, Cai R,
            Wang H, Duan J, Albarran E, Ding J, Lovinger DM, Li B, Soltesz I, Li
            Y
  TITLE     A fluorescent sensor for spatiotemporally resolved imaging of 
            endocannabinoid dynamics in vivo.
  JOURNAL   Nat Biotechnol. 2021 Nov 11. pii: 10.1038/s41587-021-01074-4. doi: 
            10.1038/s41587-021-01074-4.
  PUBMED    34764491
REFERENCE   2  (bases 1 to 7580)
  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"; journalName: "Nat 
            Biotechnol. 2021 Nov 11. pii: 10.1038/s41587-021-01074-4. doi: 
            10.1038/s41587-021-01074-4."
COMMENT     Sequence Label: pAAV-CaMKII-GRAB_eCB2.0
FEATURES             Location/Qualifiers
     source          1..7580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        145..1433
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     sig_peptide     1466..1525
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    1529..1553
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2722..2743
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2849..2870)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3110..3129)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3707..4295
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3760..3780)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4178..4189)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4327..4803
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4526..4545)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4843..4983
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4843..4972
                     /label=AAV2 ITR
     rep_origin      5058..5513
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5145..5164)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5355..5376
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5530..5549)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5649..5671
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5709..5727)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5795..5899
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5900..5968,5969..6760)
                     /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(6118..6137)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6931..7519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7420..7439
                     /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 gcgtcattat ggccttaggt cacttcatct ccatggggtt
      181 cttcttctga ttttctagaa aatgagatgg gggtgcagag agcttcctca gtgacctgcc
      241 cagggtcaca tcagaaatgt cagagctaga acttgaactc agattactaa tcttaaattc
      301 catgccttgg gggcatgcaa gtacgatata cagaaggagt gaactcatta gggcagatga
      361 ccaatgagtt taggaaagaa gagtccaggg cagggtacat ctacaccacc cgcccagccc
      421 tgggtgagtc cagccacgtt cacctcatta tagttgcctc tctccagtcc taccttgacg
      481 ggaagcacaa gcagaaactg ggacaggagc cccaggagac caaatcttca tggtccctct
      541 gggaggatgg gtggggagag ctgtggcaga ggcctcagga ggggccctgc tgctcagtgg
      601 tgacagatag gggtgagaaa gcagacagag tcattccgtc agcattctgg gtctgtttgg
      661 tacttcttct cacgctaagg tggcggtgtg atatgcacaa tggctaaaaa gcagggagag
      721 ctggaaagaa acaaggacag agacagaggc caagtcaacc agaccaattc ccagaggaag
      781 caaagaaacc attacagaga ctacaagggg gaagggaagg agagatgaat tagcttcccc
      841 tgtaaacctt agaacccagc tgttgccagg gcaacggggc aatacctgtc tcttcagagg
      901 agatgaagtt gccagggtaa ctacatcctg tctttctcaa ggaccatccc agaatgtggc
      961 acccactagc cgttaccata gcaactgcct ctttgcccca cttaatccca tcccgtctgt
     1021 taaaagggcc ctatagttgg aggtggggga ggtaggaaga gcgatgatca cttgtggact
     1081 aagtttgttc gcatcccctt ctccaacccc ctcagtacat caccctgggg gaacagggtc
     1141 cacttgctcc tgggcccaca cagtcctgca gtattgtgta tataaggcca gggcaaagag
     1201 gagcaggttt taaagtgaaa ggcaggcagg tgttggggag gcagttaccg gggcaacggg
     1261 aacagggcgt ttcggaggtg gttgccatgg ggacctggat gctgacgaag gctcgcgagg
     1321 ctgtgagcag ccacagtgcc ctgctcagaa gccccaagct cgtcagtcaa gccggttctc
     1381 cgtttgcact caggagcacg ggcaggcgag tggcccctag ttctgggggc agcggggtac
     1441 cggatcctct agagtcgact ccggaatgga gacagacaca ctcctgctat gggtactgct
     1501 gctctgggtt ccaggttcca ctggtgacac aagtttgtac aaaaaagttg gcaccaagtc
     1561 gatcctagat ggccttgcag ataccacctt ccgcaccatc accactgacc tcctgtacgt
     1621 gggctcaaat gacattcagt acgaagacat caaaggtgac atggcatcca aattagggta
     1681 cttcccacag aaattccctt taacttcctt taggggaagt cccttccaag agaagatgac
     1741 tgcgggagac aacccccagc tagtcccagc agaccaggtg aacattacag aattttacaa
     1801 caagtctctc tcgtccttca aggagaatga ggagaacatc cagtgtgggg agaacttcat
     1861 ggacatagag tgtttcatgg tcctgaaccc cagccagcag ctggccattg cagtcctgtc
     1921 cctcacgctg ggcaccttca cggtcctgga gaacctcctg gtgctgtgcg tcatcctcca
     1981 ctcccgcagc ctccgctgca ggccttccta ccacttcatc ggcagcctgg cggtggcaga
     2041 cctcctgggg agtgtcattt ttgtctacag cttcattgac ttccacgtgt tccaccgcaa
     2101 agatagccgc aacgtgtttc tgttcaaact gggtggggtc acggcctcct tcactgcctc
     2161 cgtgggcagc ctgttcctca cagccatcga caggtacata tccattcaca ggcccctggc
     2221 ctataagagg attgtcacca ggcccaaggc cgtggtggcg ttttgcctga tgtggaccat
     2281 agccattgtg atcgccgtgc tgcctctcct gggctggaac tgcgagaaac tgcaatctgt
     2341 ttgctcagac attttcccac acattgatga aacctacctg atgttctgga tcggggtcac
     2401 cagcgtactg cttctgttca tcgtgtatgc gtacatgtat attctctgga aggctcacag
     2461 ccacgccgtc cgcatgattc agcgtaccga cgcgctggac ctggaggagg gaggaaacgt
     2521 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttctgca tccgccacaa
     2581 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga
     2641 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga
     2701 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2761 tttcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gaaagggcga
     2821 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2881 caagttcagc gtgggcggcg agggtgaggg cgatgccacc gttggcaagc tgaccctgaa
     2941 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     3001 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     3061 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa
     3121 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3181 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacac
     3241 cggagcagca gcacgctggc gcgggcggcg catggacatt aggttagcca agaccctggt
     3301 cctgatcctg gtggtgttga tcatctgctg gggccctctg cttgcaatca tggtgtatga
     3361 tgtctttggg aagatgaaca agctcattaa gacggtgttt gcattctgca ccatgctctg
     3421 cctgctgaac tccaccgtga accccatcat ctatgctctg aggagtaagg acctgcgaca
     3481 cgctttccgg agcatgtttc cctcttgtga aggcactgcg cagcctctgg ataacagcat
     3541 gggggactcg gactgcctgc acaaacacgc aaacaatgca gccagtgttc acagggccgc
     3601 agaaagctgc atcaagagca cggtcaagat tgccaaggta accatgtctg tgtccacaga
     3661 cacgtctgcc gaggctctgt gagaattcga tatcaagctt atcgataatc aacctctgga
     3721 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3781 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3841 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3901 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3961 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     4021 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4081 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac
     4141 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4201 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4261 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag
     4321 atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     4381 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     4441 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     4501 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     4561 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     4621 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     4681 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     4741 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     4801 cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     4861 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4921 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     4981 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     5041 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5101 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     5161 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5221 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     5281 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5341 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     5401 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     5461 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     5521 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     5581 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5641 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5701 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5761 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5821 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5881 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5941 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6001 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6061 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     6121 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6181 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6241 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6301 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6361 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6421 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6481 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6541 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6601 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6661 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6721 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6781 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6841 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6901 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6961 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     7021 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     7081 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     7141 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7201 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7261 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7321 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7381 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7441 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7501 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7561 ctggcctttt gctcacatgt
//
