LOCUS       sequence-350417-        6293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for glutamate, third generation, variant 857. 
            iGluSnFR3.v857.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6293)
  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 6293)
  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.PDGFR
FEATURES             Location/Qualifiers
     source          1..6293
                     /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"
     CDS             3121..3267
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    3295..3883
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3348..3368)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3766..3777)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3911..4032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3922..3941)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3976..3995
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4194..4323
                     /label=AAV2 ITR
     rep_origin      complement(4560..5148)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4640..4659)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5319..6110,6111..6179))
                     /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(6180..6284)
                     /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 ggatctgaat
     3121 gctgtgggcc aggacacgca ggaggtcatc gtggtgccac actccttgcc ctttaaggtg
     3181 gtggtgatct cagccatcct ggccctggtg gtgctcacca tcatctccct tatcatcctc
     3241 atcatgcttt ggcagaagaa gccacgttag aagcttttcg aattagctta tcataatcaa
     3301 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3361 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3421 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3481 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3541 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3601 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3661 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     3721 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3781 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3841 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcggag gccgcttcga
     3901 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3961 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     4021 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     4081 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt
     4141 cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc
     4201 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4261 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4321 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4381 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4441 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4501 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4561 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4621 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4681 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4741 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4801 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4861 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4921 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4981 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5041 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5101 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5161 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5221 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5281 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5341 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5401 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5461 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5521 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5581 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5641 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5701 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5761 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5821 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5881 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5941 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6001 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6061 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6121 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6181 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6241 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tcc
//
