LOCUS       sequence-325007-        6701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the endocannabinoid non-responsive sensor GRAB_eCBmut in 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6701)
  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 6701)
  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-hSyn-GRAB_eCBmut
FEATURES             Location/Qualifiers
     source          1..6701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     587..646
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    650..674
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1843..1864
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1970..1991)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2231..2250)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2828..3416
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2881..2901)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3299..3310)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3448..3924
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3647..3666)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3964..4104
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3964..4093
                     /label=AAV2 ITR
     rep_origin      4179..4634
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4266..4285)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4476..4497
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4651..4670)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4770..4792
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4830..4848)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4916..5020
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5021..5089,5090..5881)
                     /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(5239..5258)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6052..6640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6541..6560
                     /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 gcgtatgagg cggggtgggg gtgcctacct gacgaccgac
      181 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      241 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      301 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      361 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      421 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      481 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      541 ggagtcgtgt cgtgcctgag agcgcagtcg agaaaccggt gccaccatgg agacagacac
      601 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca caagtttgta
      661 caaaaaagtt ggcaccaagt cgatcctaga tggccttgca gataccacct tccgcaccat
      721 caccactgac ctcctgtacg tgggctcaaa tgacattcag tacgaagaca tcaaaggtga
      781 catggcatcc aaattagggt acttcccaca gaaattccct ttaacttcct ttaggggaag
      841 tcccttccaa gagaagatga ctgcgggaga caacccccag ctagtcccag cagaccaggt
      901 gaacattaca gaattttaca acaagtctct ctcgtccttc aaggagaatg aggagaacat
      961 ccagtgtggg gagaacttca tggacataga gtgtttcatg gtcctgaacc ccagccagca
     1021 gctggccatt gcagtcctgt ccctcacgct gggcaccttc acggtcctgg agaacctcct
     1081 ggtgctgtgc gtcatcctcc actcccgcag cctccgctgc aggccttcct accacttcat
     1141 cggcagcctg gcggtggcag acctcctggg gagtgtcatt tttgtctaca gcttcattga
     1201 cgcccacgtg ttccaccgca aagatagccg caacgtgttt ctgttcaaac tgggtggggt
     1261 cacggcctcc ttcactgcct ccgtgggcag cctgttcctc acagccatcg acaggtacat
     1321 atccattcac aggcccctgg cctataagag gattgtcacc aggcccaagg ccgtggtggc
     1381 gttttgcctg atgtggacca tagccattgt gatcgccgtg ctgcctctcc tgggctggaa
     1441 ctgcgagaaa ctgcaatctg tttgctcaga cattttccca cacattgatg aaacctacct
     1501 gatgttctgg atcggggtca ccagcgtact gcttctgttc atcgtgtatg cgtacatgta
     1561 tattctctgg aaggctcaca gccacgccgt ccgcatgatt cagcgtaccg acgcgctgga
     1621 cctggaggag ggaggaaacg tctatatcaa ggccgacaag cagaagaacg gcatcaaggc
     1681 gaacttctgc atccgccaca acatcgagga cggcggcgtg cagctcgcct accactacca
     1741 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcgt
     1801 gcagtccaaa ctttcgaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1861 cgtgaccgcc gccgggatca ctttcggcat ggacgagctg tacaagggcg gtaccggagg
     1921 gagcatggtg agaaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1981 ggacggcgac gtaaacggcc acaagttcag cgtgggcggc gagggtgagg gcgatgccac
     2041 cgttggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2101 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2161 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacatccagg agcgcaccat
     2221 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2281 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2341 gcacaagctg gagtacaaca ccggagcagc agcacgctgg cgcgggcggc gcatggacat
     2401 taggttagcc aagaccctgg tcctgatcct ggtggtgttg atcatctgct ggggccctct
     2461 gcttgcaatc atggtgtatg atgtctttgg gaagatgaac aagctcatta agacggtgtt
     2521 tgcattctgc accatgctct gcctgctgaa ctccaccgtg aaccccatca tctatgctct
     2581 gaggagtaag gacctgcgac acgctttccg gagcatgttt ccctcttgtg aaggcactgc
     2641 gcagcctctg gataacagca tgggggactc ggactgcctg cacaaacacg caaacaatgc
     2701 agccagtgtt cacagggccg cagaaagctg catcaagagc acggtcaaga ttgccaaggt
     2761 aaccatgtct gtgtccacag acacgtctgc cgaggctctg tgagaattcg atatcaagct
     2821 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2881 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2941 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3001 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3061 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3121 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3181 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3241 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3301 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3361 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3421 ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct
     3481 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     3541 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     3601 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     3661 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     3721 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     3781 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     3841 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     3901 accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc
     3961 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4021 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4081 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     4141 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4201 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     4261 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4321 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4381 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4441 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4501 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     4561 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4621 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4681 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4741 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4801 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4861 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4921 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4981 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5041 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5101 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5161 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5221 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5281 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5341 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5401 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5461 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5521 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5581 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5641 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5701 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5761 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5821 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5881 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5941 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6001 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6061 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6121 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6181 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6241 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6301 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6361 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6421 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6481 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6541 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6601 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6661 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
