LOCUS       sequence-361545-        6056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for glutamate, third generation, variant 857 in
            GPI backbone. iGluSnFR3.v857.GPI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6056)
  AUTHORS   Aggarwal A, Liu R, Chen Y, Ralowicz AJ, Bergerson SJ, Tomaska F, 
            Mohar B, Hanson TL, Hasseman JP, Reep D, Tsegaye G, Yao P, Ji X, 
            Kloos M, Walpita D, Patel R, Mohr MA, Tillberg PW, Looger LL, Marvin
            JS, Hoppa MB, Konnerth A, Kleinfeld D, Schreiter ER, Podgorski K
  TITLE     Glutamate indicators with improved activation kinetics and 
            localization for imaging synaptic transmission.
  JOURNAL   Nat Methods. 2023 May 4. doi: 10.1038/s41592-023-01863-6.
  PUBMED    37142767
REFERENCE   2  (bases 1 to 6056)
  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 
            Methods. 2023 May 4. doi: 10.1038/s41592-023-01863-6."
COMMENT     Sequence Label: pAAV.hSyn.iGluSnFR3.v857.GPI
FEATURES             Location/Qualifiers
     source          1..6056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(317..336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(996..1787,1788..1856))
                     /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     1619..1638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1857..1961)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1987..2442)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2124..2145)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2336..2355
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2514..2643
                     /label=AAV2 ITR
     promoter        2719..3166
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3195..3204
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3201..3260
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             4845..4874
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    5007..5595
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5060..5080)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5478..5489)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5598..5614)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5599..5615)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5644..5765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5655..5674)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5709..5728
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5927..6056
                     /label=AAV2 ITR
ORIGIN
        1 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
       61 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      661 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1201 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1321 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1381 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1441 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1621 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1861 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1921 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1981 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2041 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2101 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2161 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2221 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2281 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2341 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2401 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     2461 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgcgcg
     2521 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
     2581 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     2641 ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga
     2701 tccagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg
     2761 tgcctacctg acgaccggcc ccgacccact ggacaagcac ccaaccccca ttccccaaat
     2821 tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact
     2881 gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg
     2941 cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg
     3001 ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga
     3061 gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca
     3121 gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcca attcaagctg
     3181 ctagcaagga tcccgccacc atggagacag acacactcct gctatgggta ctgctgctct
     3241 gggttccagg ttccactggt gacagatctg ggtccgccgc cggaagtact cttgataaga
     3301 tcgctaaaaa tggtgtaatt gttgtcggcc accgagactc ttctgtgcca ttcagctatt
     3361 atgattctca acacaaagtg gtgggctatt cacaagagta ttctaacgcc attgttgaag
     3421 cagttaagaa gaagttgaat aaaccagacc ttcaagtaaa actcattcct ataacatccc
     3481 aaaaccgcat acctctcttg caaaacggca cttacgattt cgagtgtggg agcactacta
     3541 ataatgtcga acgccagaaa caagccgcat tctccgacac catcttcgtg gtgggcactc
     3601 gactgttgac aaagaaaggc ggtgatatta aggatttcgc aaatctgaag gataaggcag
     3661 tcgtggtcac cagcgggacc acctcagaag tactccttaa taaactcaat gaagaacaaa
     3721 agatgaatat gcgaatcatc tctgctaagg accacggtga ctcttttcgc acattggaaa
     3781 gtgggcgcgc cgtagcattt atgatggacg actctctgct cgctggcgag cgcgccaaag
     3841 ccaaaaaacc agacgactgg gaaattgtag gtaaacctca aagccaagag gcatatggct
     3901 gcatgttgag aaaagatgac ccccaattca agaaactgat ggacgacact atagctcagg
     3961 tccagacttc aggggaagct gagaagtggt ttgataaatg gttcaagaac ccaatcttgg
     4021 taaatcatac cgtttatatt accgctgaca aacaaaagaa tgggataaag gcaaatttca
     4081 agattcacca taacgtagag gatggaagcg tgcagttggc agaccattat caacagaata
     4141 cccctattgg cgacggaccc gtgctcctcc ctgacaacca ctacttgtcc tatcaaagtg
     4201 ttctctcaaa ggaccccaat gagaagcgag accacatggt acttctcgaa ttcgttaccg
     4261 ccaccgggaa ttcacttggc atggatgaat tgtataaggg gggtactggc ggcagcatgt
     4321 ccaagggtga ggagcttttt accggagtag taccaattct cgttgaactt gacggagacg
     4381 tcaacgggca caagtttagc gtacgaggtg agggggaggg tgatgcaacc aacggtaaac
     4441 ttactctcaa attcatatgc actaccggaa agctgccagt accttggcca acactggtca
     4501 ctaccctctc ctatggcgtt caatgttttg ctcgataccc tgaccacatg aaacagcatg
     4561 acttctttaa atcagccatg cctgaagggt atgtccagga gcggactata tcatttaaag
     4621 acgatggaac ctataagact cgggctgaag tgaagctcga gggtgacaca ttggttaatc
     4681 ggatcgagct taagggcata gattttaaag aagatggtaa tattcttggc cacaaattgg
     4741 agtacaattt taataatcct ttgaacatga actttgagct ctccgatgaa atgaaggcac
     4801 tctttaaaga gccaaacgac aaagctctga aactgcaggt cgacgaacaa aaactcatct
     4861 cagaagagga tctgtcagat aacgactacg gtaaagatgg gttggctggg gcttcaggag
     4921 gcagacgaaa tggcgctata actgtgctca gtttcataac gttctacgtg gcagcattta
     4981 tggtattgtt gtagaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     5041 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     5101 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     5161 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     5221 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     5281 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     5341 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     5401 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     5461 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     5521 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     5581 gggccgcctc cccgcatcga taccgtcgac ctcgacccgg gcggccgctt cgagcagaca
     5641 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     5701 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     5761 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     5821 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag
     5881 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
     5941 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     6001 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcag
//
