LOCUS       sequence-326663-        6820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV for glutamate reporter fused to TARP gamma-2 for postsynaptic 
            localisation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6820)
  AUTHORS   Hao Y, Toulme E, Konig B, Rosenmund C, Plested AJR
  TITLE     Targeted sensors for glutamatergic neurotransmission.
  JOURNAL   eLife. 2023 Jan 9;12:e84029. doi: 10.7554/eLife.84029.
  PUBMED    36622100
REFERENCE   2  (bases 1 to 6820)
  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: "eLife."; 
            date: "2023-01-9"; volume: "12:e84029. doi"; pages: " 
            10.7554/eLife.84029"
COMMENT     Sequence Label: pAAV-syn-SnFR-gamma2-minWPRE
FEATURES             Location/Qualifiers
     source          1..6820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     sig_peptide     92..151
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1120..1141
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1247..1268)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1508..1527)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1738..1756
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     complement(3078..3098)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3474..3614
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3474..3603
                     /label=AAV2 ITR
     rep_origin      3689..4144
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3776..3795)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3986..4007
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4161..4180)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4280..4302
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4340..4358)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4426..4530
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4531..4599,4600..5391)
                     /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(4749..4768)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5562..6150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6051..6070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6212..6341
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        6373..6820
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 tcgctagcgt ttaaacttaa gcttggtacc gaactcggat ccactagtaa cggccgccag
       61 tgtgctggaa ttcggcttgg ggatatccac catggagaca gacacactcc tgctatgggt
      121 actgctgctc tgggttccag gttccactgg tgacagatct gccgcaggca gcacgctgga
      181 caaaatcgcc aaaaacggtg tgattgtcgt cggtcaccgt gaatcttcag tgcctttctc
      241 ttattacgac aatcagcaaa aagtggtggg ttactcgcag gattactcca acgccattgt
      301 tgaagcagtg aaaaagaaac tcaacaaacc ggacttgcag gtaaaactga ttccgattac
      361 ctcacaaaac cgtattccac tgctgcaaaa cggcactttc gattttgaat gtggttctac
      421 caccaacaac gtcgaacgcc aaaaacaggc ggctttctct gacactattt tcgtggtcgg
      481 tacgcgcctg ttgaccaaaa agggtggcga tatcaaagat tttgccaacc tgaaagacaa
      541 agccgtagtc gtcacttccg gcactacctc tgaagttttg ctcaacaaac tgaatgaaga
      601 gcaaaaaatg aatatgcgca tcatcagcgc caaagatcac ggtgactctt tccgcaccct
      661 ggaaagcggt cgtgccgttg cctttatgat ggatgacgtg ctgctggccg gtgaacgtgc
      721 gaaagcgaag aaaccagaca actgggaaat cgtcggcaag ccgcagtctc aggaggccta
      781 cggttgtatg ttgcgtaaag atgatccgca gttcaaaaag ctgatggatg acaccatcgc
      841 tcaggtgcag acctccggtg aagcggaaaa atggtttgat aagtggttca aaaatccaat
      901 tctggtgagc cacaacgtct atatcatggc cgacaagcag aggaacggca tcaaggcgaa
      961 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc actaccagca
     1021 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1081 gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1141 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggta ccggagggag
     1201 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1261 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggtgagggcg atgccaccta
     1321 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1381 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     1441 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac atccaggagc gcaccatctt
     1501 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1561 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1621 taagctggag tacaacttta acaaccccct gaacatgaat ttcgaactgt cagacgaaat
     1681 gaaagcactg ttcaaagaac cgaatgacaa ggcactgaaa ggtgggcgcg cccgggcaga
     1741 tgtgtataag agacaggaaa agaaaaaagc aggactgttt gaacaaatca cgaagactca
     1801 tgggacgatc attggtgtca catcagggat cgtcctggtc cttctcatta tttctatttt
     1861 agtacaagtg aaacagcccc ggaaaaaggt catggcttgc aaaactgcat ttaataaaac
     1921 tgggttccag gaagtatttg atcctcctca ttatgaacta ttttcactaa gagagaaaga
     1981 gatttctgca gacgccgggt ccgggctgtt tgatcgaggt gttcaaatgc ttttaaccac
     2041 cgttggtgct ttcgctgcct tcagcttgat gaccatcgct gtgggaaccg actattggct
     2101 gtactccaga ggggtttgca agaccaaaag tgtcagtgag aatgaaacca gcaagaagaa
     2161 cgaggaagtt atgacccatt ccggattatg gagaacctgc tgcctcgaag ggaacttcaa
     2221 aggtctgtgc aagcaaatcg accactttcc ggaagacgcg gactacgaag ctgacaccgc
     2281 agagtatttc ctccgggccg tgagggcctc gagtatcttc ccgatcctga gtgtgatcct
     2341 gcttttcatg ggtggcctct gcatcgcggc gagcgagttc tacaagacac gccacaacat
     2401 catcctgagt gctggcatct tcttcgtgtc tgcaggtctt agtaatatca tcgggatcat
     2461 cgtgtatata tcagccaatg ctggagaccc ctccaagagt gactccaaaa agaacagcta
     2521 ctcctacggc tggtccttct acttcggggc cctgtccttc atcatcgccg agatggtcgg
     2581 ggtgctggcc gtgcacatgt ttatcgaccg ccacaaacag ctgcgggcca cggcccgcgc
     2641 caccgactac ctccaggcct ccgccatcac ccgcatcccc agctaccgct accgctacca
     2701 gcgccgcagc cgctccagct cgcgctccac cgagccctct cactccagag acgcctcgcc
     2761 cgtgggcgtg aagggcttca acaccctgcc gtccacggag atctccatgt acaccctcag
     2821 tagggacccc ctgaaggctg ccaccacgcc caccgccacc tacaactcgg acagggataa
     2881 cagctttctc caggtccaca actgtatcca gaaggacagc aaggactctc tccacgccaa
     2941 cacagccaac cgccggacca cgcccgtatg agaattctgc agatatccag cacagtggcg
     3001 gccgctcgag tctagactcg agataatcaa cctctggatt acaaaatttg tgaaagattg
     3061 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3121 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3181 ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3241 cggctgttgg gcactgacaa ttccgtggtg tttatttgtg aaatttgtga tgctattgct
     3301 ttatttgtaa ccatctagct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3361 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3421 tcagggggag atgtgggagg ttttttaaag cacggtgcgg accgagcggc cgcaggaacc
     3481 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     3541 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     3601 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3661 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     3721 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     3781 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3841 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3901 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3961 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4021 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     4081 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4141 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4201 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4261 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4321 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4381 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4441 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4501 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4561 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4621 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4681 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4741 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4801 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4861 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4921 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4981 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5041 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5101 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5161 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5221 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5281 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5341 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5401 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5461 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5521 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5581 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5641 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5701 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5761 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5821 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5881 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5941 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6001 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6061 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6121 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6181 ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc
     6241 actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     6301 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca cgcgtagagg
     6361 gccctgcgta tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta
     6421 cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca
     6481 tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc
     6541 ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag
     6601 cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct
     6661 tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg
     6721 ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc
     6781 ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag
//
