LOCUS       sequence-405482-        6814 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 6814)
  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 6814)
  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.CAG.FLEX.iGluSnFR3.v857.GPI
FEATURES             Location/Qualifiers
     source          1..6814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1133..1166
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1237..1270
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      1278..1287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1284..1343
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2928..2957
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(3090..3123)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3186..3219)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(3257..3276)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3338..3360)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3639..3916)
                     /label=chicken beta-actin promoter
     repeat_region   4227..4356
                     /label=AAV2 ITR
     rep_origin      4428..4883
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4515..4534)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4725..4746
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4909..5013
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5014..5082,5083..5874)
                     /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(5232..5251)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6045..6633
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6534..6553
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct aattcgaacc tcttcgaggg acataacttc
     1141 gtataatgta tgctatacga agttatgggt cgatggtgag atctggacta gagggtcgat
     1201 ggtgatgctt ggatcaaaat aggaagaatc ccaatgataa cttcgtataa agtatcctat
     1261 acgaagttat ggatcccgcc accatggaga cagacacact cctgctatgg gtactgctgc
     1321 tctgggttcc aggttccact ggtgacagat ctgggtccgc cgccggaagt actcttgata
     1381 agatcgctaa aaatggtgta attgttgtcg gccaccgaga ctcttctgtg ccattcagct
     1441 attatgattc tcaacacaaa gtggtgggct attcacaaga gtattctaac gccattgttg
     1501 aagcagttaa gaagaagttg aataaaccag accttcaagt aaaactcatt cctataacat
     1561 cccaaaaccg catacctctc ttgcaaaacg gcacttacga tttcgagtgt gggagcacta
     1621 ctaataatgt cgaacgccag aaacaagccg cattctccga caccatcttc gtggtgggca
     1681 ctcgactgtt gacaaagaaa ggcggtgata ttaaggattt cgcaaatctg aaggataagg
     1741 cagtcgtggt caccagcggg accacctcag aagtactcct taataaactc aatgaagaac
     1801 aaaagatgaa tatgcgaatc atctctgcta aggaccacgg tgactctttt cgcacattgg
     1861 aaagtgggcg cgccgtagca tttatgatgg acgactctct gctcgctggc gagcgcgcca
     1921 aagccaaaaa accagacgac tgggaaattg taggtaaacc tcaaagccaa gaggcatatg
     1981 gctgcatgtt gagaaaagat gacccccaat tcaagaaact gatggacgac actatagctc
     2041 aggtccagac ttcaggggaa gctgagaagt ggtttgataa atggttcaag aacccaatct
     2101 tggtaaatca taccgtttat attaccgctg acaaacaaaa gaatgggata aaggcaaatt
     2161 tcaagattca ccataacgta gaggatggaa gcgtgcagtt ggcagaccat tatcaacaga
     2221 atacccctat tggcgacgga cccgtgctcc tccctgacaa ccactacttg tcctatcaaa
     2281 gtgttctctc aaaggacccc aatgagaagc gagaccacat ggtacttctc gaattcgtta
     2341 ccgccaccgg gaattcactt ggcatggatg aattgtataa ggggggtact ggcggcagca
     2401 tgtccaaggg tgaggagctt tttaccggag tagtaccaat tctcgttgaa cttgacggag
     2461 acgtcaacgg gcacaagttt agcgtacgag gtgaggggga gggtgatgca accaacggta
     2521 aacttactct caaattcata tgcactaccg gaaagctgcc agtaccttgg ccaacactgg
     2581 tcactaccct ctcctatggc gttcaatgtt ttgctcgata ccctgaccac atgaaacagc
     2641 atgacttctt taaatcagcc atgcctgaag ggtatgtcca ggagcggact atatcattta
     2701 aagacgatgg aacctataag actcgggctg aagtgaagct cgagggtgac acattggtta
     2761 atcggatcga gcttaagggc atagatttta aagaagatgg taatattctt ggccacaaat
     2821 tggagtacaa ttttaataat cctttgaaca tgaactttga gctctccgat gaaatgaagg
     2881 cactctttaa agagccaaac gacaaagctc tgaaactgca ggtcgacgaa caaaaactca
     2941 tctcagaaga ggatctgtca gataacgact acggtaaaga tgggttggct ggggcttcag
     3001 gaggcagacg aaatggcgct ataactgtgc tcagtttcat aacgttctac gtggcagcat
     3061 ttatggtatt gttgtagaag cttgctagta taacttcgta tagcatacat tatacgaagt
     3121 tatgggtcga tggtgatgct tggcaattcg ggtcgatggt gaagcattgg tcttcctatt
     3181 ttgatataac ttcgtatagg atactttata cgaagttatg gggatgatcc aattctttgc
     3241 caaaatgatg agacagcaca ataaccagca cgttgcccag gagctgtagg aaaaagaaga
     3301 aggcatgaac atggttagca gaggctctag agccgccggt cacacgccag aagccgaacc
     3361 ccgccctgcc ccgtcccccc cgaaggcagc cgtccccccg cggacagccc cgagccgctc
     3421 cccccgcaca aagggccctc ccggagcccc tcaaggcttt cacgcagcca cagaaaagaa
     3481 acaagccgtc attaaaccaa gcgctaatta cagcccggag gagaagggcc gtcccgcccg
     3541 ctcacctgtg ggagtaacgc ggtcagtcag agccggggcg ggcggcgcga ggcggcggcg
     3601 gagcggggca cggggcgaag gcagcgcgca gcgactcccg cccgccgcgc gcttcgcttt
     3661 ttatagggcc gccgccgccg ccgcctcgcc ataaaaggaa actttcggag cgcgccgctc
     3721 tgattggctg ccgccgcacc tctccgcctc gccccgcccc gcccctcgcc ccgccccgcc
     3781 ccgcctggcg cgcgcccccc cccccccccc gcccccatcg ctgcacaaaa taattaaaaa
     3841 ataaataaat acaaaattgg gggtggggag gggggggaga tggggagagt gaagcagaac
     3901 gtggggctca cctcgaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     3961 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     4021 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     4081 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     4141 acatacgtca ttattgacgt caatggtact cttcctagag catggctacg tagataagta
     4201 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     4261 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4321 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgg cgtaatagcg aagaggcccg
     4381 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     4441 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4501 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4561 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4621 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4681 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4741 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4801 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4861 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     4921 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4981 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5041 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5101 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5161 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5221 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5281 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5341 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5401 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5461 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5521 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5581 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5641 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5701 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5761 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5821 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5881 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5941 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6001 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6061 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6121 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6181 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6241 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6301 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6361 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6421 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6481 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6541 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6601 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6661 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6721 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6781 gagcgcagcg agtcagtgag cgaggaagcg gaag
//
