LOCUS       sequence-322167-        6830 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 6830)
  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 6830)
  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..6830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        173..620
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     sig_peptide     712..771
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1740..1761
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1867..1888)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2128..2147)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2358..2376
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     complement(3698..3718)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4093..4233
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4093..4222
                     /label=AAV2 ITR
     rep_origin      4308..4763
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4395..4414)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4605..4626
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4780..4799)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4899..4921
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4959..4977)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5045..5149
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5150..5218,5219..6010)
                     /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(5368..5387)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6181..6769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6670..6689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtagagg gccctgcgta tgagtgcaag
      181 tgggttttag gaccaggatg aggcggggtg ggggtgccta cctgacgacc gaccccgacc
      241 cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca gagaggggga
      301 ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc
      361 cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt
      421 cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc cgcgccgccg
      481 ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc
      541 tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg
      601 tgtcgtgcct gagagcgcag tcgctagcgt ttaaacttaa gcttggtacc gaactcggat
      661 ccactagtaa cggccgccag tgtgctggaa ttcggcttgg ggatatccac catggagaca
      721 gacacactcc tgctatgggt actgctgctc tgggttccag gttccactgg tgacagatct
      781 gccgcaggca gcacgctgga caaaatcgcc aaaaacggtg tgattgtcgt cggtcaccgt
      841 gaatcttcag tgcctttctc ttattacgac aatcagcaaa aagtggtggg ttactcgcag
      901 gattactcca acgccattgt tgaagcagtg aaaaagaaac tcaacaaacc ggacttgcag
      961 gtaaaactga ttccgattac ctcacaaaac cgtattccac tgctgcaaaa cggcactttc
     1021 gattttgaat gtggttctac caccaacaac gtcgaacgcc aaaaacaggc ggctttctct
     1081 gacactattt tcgtggtcgg tacgcgcctg ttgaccaaaa agggtggcga tatcaaagat
     1141 tttgccaacc tgaaagacaa agccgtagtc gtcacttccg gcactacctc tgaagttttg
     1201 ctcaacaaac tgaatgaaga gcaaaaaatg aatatgcgca tcatcagcgc caaagatcac
     1261 ggtgactctt tccgcaccct ggaaagcggt cgtgccgttg cctttatgat ggatgacgtg
     1321 ctgctggccg gtgaacgtgc gaaagcgaag aaaccagaca actgggaaat cgtcggcaag
     1381 ccgcagtctc aggaggccta cggttgtatg ttgcgtaaag atgatccgca gttcaaaaag
     1441 ctgatggatg acaccatcgc tcaggtgcag acctccggtg aagcggaaaa atggtttgat
     1501 aagtggttca aaaatccaat tctggtgagc cacaacgtct atatcatggc cgacaagcag
     1561 aggaacggca tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     1621 ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1681 aaccactacc tgagcaccca gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac
     1741 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1801 aagggcggta ccggagggag catggtgagc aagggcgagg agctgttcac cggggtggtg
     1861 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1921 ggtgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1981 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2041 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2101 atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2161 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2221 gacggcaaca tcctggggca taagctggag tacaacttta acaaccccct gaacatgaat
     2281 ttcgaactgt cagacgaaat gaaagcactg ttcaaagaac cgaatgacaa ggcactgaaa
     2341 ggtgggcgcg cccgggcaga tgtgtataag agacaggaaa agaaaaaagc aggactgttt
     2401 gaacaaatca cgaagactca tgggacgatc attggtgtca catcagggat cgtcctggtc
     2461 cttctcatta tttctatttt agtacaagtg aaacagcccc ggaaaaaggt catggcttgc
     2521 aaaactgcat ttaataaaac tgggttccag gaagtatttg atcctcctca ttatgaacta
     2581 ttttcactaa gagagaaaga gatttctgca gacgccgggt ccgggctgtt tgatcgaggt
     2641 gttcaaatgc ttttaaccac cgttggtgct ttcgctgcct tcagcttgat gaccatcgct
     2701 gtgggaaccg actattggct gtactccaga ggggtttgca agaccaaaag tgtcagtgag
     2761 aatgaaacca gcaagaagaa cgaggaagtt atgacccatt ccggattatg gagaacctgc
     2821 tgcctcgaag ggaacttcaa aggtctgtgc aagcaaatcg accactttcc ggaagacgcg
     2881 gactacgaag ctgacaccgc agagtatttc ctccgggccg tgagggcctc gagtatcttc
     2941 ccgatcctga gtgtgatcct gcttttcatg ggtggcctct gcatcgcggc gagcgagttc
     3001 tacaagacac gccacaacat catcctgagt gctggcatct tcttcgtgtc tgcaggtctt
     3061 agtaatatca tcgggatcat cgtgtatata tcagccaatg ctggagaccc ctccaagagt
     3121 gactccaaaa agaacagcta ctcctacggc tggtccttct acttcggggc cctgtccttc
     3181 atcatcgccg agatggtcgg ggtgctggcc gtgcacatgt ttatcgaccg ccacaaacag
     3241 ctgcgggcca cggcccgcgc caccgactac ctccaggcct ccgccatcac ccgcatcccc
     3301 agctaccgct accgctacca gcgccgcagc cgctccagct cgcgctccac cgagccctct
     3361 cactccagag acgcctcgcc cgtgggcgtg aagggcttca acaccctgcc gtccacggag
     3421 atctccatgt acaccctcag tagggacccc ctgaaggctg ccaccacgcc caccgccacc
     3481 tacaactcgg acagggataa cagctttctc caggtccaca actgtatcca gaaggacagc
     3541 aaggactctc tccacgccaa cacagccaac cgccggacca cgcccgtatg agaattctgc
     3601 agatatccag cacagtggcg gccgctcgag tctagactcg agataatcaa cctctggatt
     3661 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3721 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3781 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     3841 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg tttatttgtg
     3901 aaatttgtga tgctattgct ttatttgtaa ccatctagct ttatttgtga aatttgtgat
     3961 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     4021 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag cacgtgcgga
     4081 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4141 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4201 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     4261 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     4321 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4381 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4441 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4501 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     4561 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4621 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     4681 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4741 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     4801 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4861 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4921 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     4981 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5041 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5101 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5161 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5221 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5281 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5341 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5401 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5461 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5521 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5581 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5641 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5701 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5761 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5821 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5881 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5941 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6001 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6061 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6121 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6181 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6241 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6301 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     6361 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6421 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6481 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6541 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6601 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6661 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6721 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6781 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
