LOCUS       sequence-361534-        6260 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 6260)
  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 6260)
  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.GFAP.iGluSnFR3.v857.GPI
FEATURES             Location/Qualifiers
     source          1..6260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(17..472)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(154..175)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     366..385
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   544..673
                     /label=AAV2 ITR
     regulatory      1438..1447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1444..1503
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             3088..3117
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    3262..3850
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3315..3335)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3733..3744)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3878..3999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3889..3908)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3943..3962
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4161..4290
                     /label=AAV2 ITR
     rep_origin      complement(4527..5115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4607..4626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5286..6077,6078..6146))
                     /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"
     promoter        complement(6147..6251)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
       61 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
      121 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
      181 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
      241 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
      301 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
      361 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
      421 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
      481 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      541 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
      601 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      661 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      721 tctaggaaga tcgtaggtta attaataaca tatcctggtg tggagtaggg gacgctgctc
      781 tgacagaggc tcgggggcct gagctggctc tgtgagctgg ggaggaggca gacagccagg
      841 cctttgtctg caagcagacc tggcagcatt gggctggccg ccccccaggg cctcctcttc
      901 atgcccagtg aatgactcac cttggcacag acacaatgtt cggggtgggc acagtgcctg
      961 cttcccgccg caccccagcc cccctcaaat gccttccgag aagcccattg agcagggggc
     1021 ttgcattgca ccccagcctg acagcctggc atcttgggat aaaagcagca cagcccccta
     1081 ggggctgccc ttgctgtgtg gcgccaccgg cggtggagaa caaggctcta ttcagcctgt
     1141 gcccaggaaa ggggatcagg ggatgcccag gcatggacag tgggtggcag ggggggagag
     1201 gagggctgtc tgcttcccag aagtccaagg acacaaatgg gtgaggggag agctctcccc
     1261 atagctgggc tgcggcccaa ccccaccccc tcaggctatg ccagggggtg ttgccagggg
     1321 cacccgggca tcgccagtct agcccactcc ttcataaagc cctcgcatcc caggagcgag
     1381 cagagccaga gcaggttgga gaggagacgc atcacctccg ctgctcgcgg ggatcccgcc
     1441 accatggaga cagacacact cctgctatgg gtactgctgc tctgggttcc aggttccact
     1501 ggtgacagat ctgggtccgc cgccggaagt actcttgata agatcgctaa aaatggtgta
     1561 attgttgtcg gccaccgaga ctcttctgtg ccattcagct attatgattc tcaacacaaa
     1621 gtggtgggct attcacaaga gtattctaac gccattgttg aagcagttaa gaagaagttg
     1681 aataaaccag accttcaagt aaaactcatt cctataacat cccaaaaccg catacctctc
     1741 ttgcaaaacg gcacttacga tttcgagtgt gggagcacta ctaataatgt cgaacgccag
     1801 aaacaagccg cattctccga caccatcttc gtggtgggca ctcgactgtt gacaaagaaa
     1861 ggcggtgata ttaaggattt cgcaaatctg aaggataagg cagtcgtggt caccagcggg
     1921 accacctcag aagtactcct taataaactc aatgaagaac aaaagatgaa tatgcgaatc
     1981 atctctgcta aggaccacgg tgactctttt cgcacattgg aaagtgggcg cgccgtagca
     2041 tttatgatgg acgactctct gctcgctggc gagcgcgcca aagccaaaaa accagacgac
     2101 tgggaaattg taggtaaacc tcaaagccaa gaggcatatg gctgcatgtt gagaaaagat
     2161 gacccccaat tcaagaaact gatggacgac actatagctc aggtccagac ttcaggggaa
     2221 gctgagaagt ggtttgataa atggttcaag aacccaatct tggtaaatca taccgtttat
     2281 attaccgctg acaaacaaaa gaatgggata aaggcaaatt tcaagattca ccataacgta
     2341 gaggatggaa gcgtgcagtt ggcagaccat tatcaacaga atacccctat tggcgacgga
     2401 cccgtgctcc tccctgacaa ccactacttg tcctatcaaa gtgttctctc aaaggacccc
     2461 aatgagaagc gagaccacat ggtacttctc gaattcgtta ccgccaccgg gaattcactt
     2521 ggcatggatg aattgtataa ggggggtact ggcggcagca tgtccaaggg tgaggagctt
     2581 tttaccggag tagtaccaat tctcgttgaa cttgacggag acgtcaacgg gcacaagttt
     2641 agcgtacgag gtgaggggga gggtgatgca accaacggta aacttactct caaattcata
     2701 tgcactaccg gaaagctgcc agtaccttgg ccaacactgg tcactaccct ctcctatggc
     2761 gttcaatgtt ttgctcgata ccctgaccac atgaaacagc atgacttctt taaatcagcc
     2821 atgcctgaag ggtatgtcca ggagcggact atatcattta aagacgatgg aacctataag
     2881 actcgggctg aagtgaagct cgagggtgac acattggtta atcggatcga gcttaagggc
     2941 atagatttta aagaagatgg taatattctt ggccacaaat tggagtacaa ttttaataat
     3001 cctttgaaca tgaactttga gctctccgat gaaatgaagg cactctttaa agagccaaac
     3061 gacaaagctc tgaaactgca ggtcgacgaa caaaaactca tctcagaaga ggatctgtca
     3121 gataacgact acggtaaaga tgggttggct ggggcttcag gaggcagacg aaatggcgct
     3181 ataactgtgc tcagtttcat aacgttctac gtggcagcat ttatggtatt gttgtagaag
     3241 cttttcgaat tagcttatca taatcaacct ctggattaca aaatttgtga aagattgact
     3301 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3361 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3421 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3481 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3541 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3601 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3661 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3721 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3781 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3841 gcctccccgc atcggaggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     3901 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3961 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     4021 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac
     4081 aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg
     4141 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
     4201 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     4261 ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg
     4321 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4381 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4441 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4501 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4561 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4621 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4681 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4741 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4801 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4861 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4921 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4981 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5041 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5101 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5161 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5221 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5281 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5341 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5401 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5461 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5521 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5581 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5641 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5701 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5761 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5821 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5881 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5941 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6001 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6061 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6121 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6181 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6241 ggggttccgc gcacatttcc
//
