LOCUS       sequence-345962-        6640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for glutamate, third generation, variant 82 in 
            GPI backbone. iGluSnFR3.v82.GPI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6640)
  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 6640)
  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.FLEX.iGluSnFR3.v82.GPI.codonopt
FEATURES             Location/Qualifiers
     source          1..6640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      69..524
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(156..175)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     366..387
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        550..654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(655..723,724..1515)
                     /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(873..892)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1686..2274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2175..2194
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2511..2640
                     /label=AAV2 ITR
     polyA_signal    2802..2923
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2839..2858)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2893..2912
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3001..3589
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3107..3118
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3516..3536
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3661..3694
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3765..3798
                     /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      3823..3832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3829..3888
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     4857..4878
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4981..5002)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             5467..5496
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(5641..5674)
                     /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(5737..5770)
                     /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(5781..5800)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          5783..5915
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   6508..6637
                     /label=AAV2 ITR
ORIGIN
        1 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
       61 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
      121 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
      181 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      241 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
      301 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      361 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
      421 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
      481 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
      541 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      601 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      661 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      721 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      781 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      841 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      901 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
      961 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1021 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1081 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1141 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1201 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1261 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1321 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1381 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1441 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1501 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1561 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1621 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1681 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1741 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1801 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1861 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1921 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1981 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2041 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2101 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2161 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2221 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2281 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2341 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2401 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2461 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctgcgcgctc
     2521 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg
     2581 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     2641 tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct aggaagatcc
     2701 ttatcgattt taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacatctcc
     2761 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2821 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2881 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgctcgaagc
     2941 ggccgcccgg gtcgactcta gatgcatgct cgagcggccg cccgggtcga cggtatcgat
     3001 gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga agacgcggaa
     3061 gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg agggccgaag
     3121 ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacacaggcg agcagccaag
     3181 gaaaggacga tgatttcccc gacaacacca cggaattgtc agtgcccaac agccgagccc
     3241 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata gggaggggga
     3301 aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca gtgggggttg
     3361 cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac aactcctcat
     3421 aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
     3481 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
     3541 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta tcgataagct
     3601 tctcgagtgg atcatctgca gaattcgccc ttttcagtgt ttcgaacctc ttcgagggac
     3661 ataacttcgt ataatgtatg ctatacgaag ttatgggtcg atggtgagat ctggactaga
     3721 gggtcgatgg tgatgcttgg atcaaaatag gaagaatccc aatgataact tcgtataaag
     3781 tatcctatac gaagttatac tagtggtacc accggtgata tcgccaccat ggagacagac
     3841 acactcctgc tatgggtact gctgctctgg gttccaggtt ccactggtga cggatccgcc
     3901 gcaggcagca cgctggacaa aatcgccaaa aacggtgtga ttgtcgtcgg tcaccgtgac
     3961 tcttcagtgc ctttctctta ttacgacagt cagcacaaag tggtgggtta ctcgcaggaa
     4021 tactccaacg ccattgttga agcagtgaaa aagaaactca acaaaccgga cttgcaggta
     4081 aaactgattc cgattacttc acaaaaccgt attccactac tgcaaaacgg cacttacgat
     4141 tttgaatgtg gttctaccac caacaacgtc gaacgccaaa aacaggcggc tttctctgac
     4201 actattttcg tggtcggtac gcgcctgttg accaaaaagg gtggcgatat caaagatttt
     4261 gccaacctga aagacaaagc cgtagtcgtc acttccggca ctacctctga agttttgctc
     4321 aacaaactga atgaagagca aaaaatgaat atgcgcatca tcagcgccaa agatcacggt
     4381 gactctttcc gcaccctgga aagcggtcgt gccgttgcct ttatgatgga tgactcgctg
     4441 ctggccggtg aacgtgcgaa agcgaagaaa ccagacgact gggaaatcgt cggcaagccg
     4501 cagtctcagg aggcctacgg ttgtatgttg cgtaaagatg atccgcagtt caaaaagctg
     4561 atggatgaca ccatcgctca ggtgcagacc tccggtgaag cggaaaaatg gtttgataag
     4621 tggttcaaaa atccaattct ggtgaaccac atcgtctata tcaccgccga caagcagaag
     4681 aacggcatca aggcgaactt caagatccac cacaacgtgg aggacggcag cgtgcagctc
     4741 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     4801 cactacctga gctatcagtc cgtgctgagc aaagacccca acgagaagcg cgatcacatg
     4861 gtcctgctgg agttcgtgac cgccaccggg aactctctcg gcatggacga gctgtacaag
     4921 ggcggtaccg gagggagcat gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     4981 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgcgcgg cgagggcgag
     5041 ggcgatgcca ccaacggcaa gctaaccctg aagctcatct gcaccaccgg caagctgccc
     5101 gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac
     5161 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     5221 gagcgcacca tcagcttcaa ggacgacgga acctacaaga cccgcgccga ggtgaagtta
     5281 gagggcgaca ccctggtgaa ccgcatcgag ctaaagggca tcgacttcaa ggaggacggc
     5341 aacatcctgg ggcacaagct ggagtacaac tttaacaacc ccctgaacat gaatttcgaa
     5401 ctgtcagacg aaatgaaagc actgttcaaa gaaccgaatg acaaggcact gaaactgcag
     5461 gtcgacgaac aaaaactcat ctcagaagag gatctgtcag ataacgacta cggtaaagat
     5521 gggttggctg gggcttccgg aggcagacga aatggcgcta taactgtgct cagtttcata
     5581 acgttctacg tggcagcatt tatggtattg ttgtaggata tcggatccgc tagcactagt
     5641 ataacttcgt atagcataca ttatacgaag ttatgtgtcg atggtgatgc ttggcaattc
     5701 gggtcgatgg tgaagcattg gtcttcctat tttgatataa cttcgtatag gatactttat
     5761 acgaagttat ggggatcatc acctgtggag agaaaggcaa agtggatgtc agtaagacca
     5821 ataggtgcct atcagaaacg caagagtctt ctctgtctcg acaagcccag tttctattgg
     5881 tctccttaaa cctgtcttgt aaccttgata cttacctgat ccaacggctg cgacttgggg
     5941 cagggggtcc taggggtggt ctggccagga gccgcggggg cggactgcgc ggtgcccagg
     6001 agcacagctg cgctctcagg cacgacacga ctcctccgct gcccaccgca gactgaggca
     6061 gcgctgagtc gccggcgccg cagcgcagat ggtcgcgccc gtgcccccct atctcgcgcc
     6121 tcgcgtggtg cggtccggct gggccggcgg cggcgcggac gcgaccaagg tggccgggaa
     6181 ggggagtttg cgggggaccg gcgagtgacg tcagcgcgcc ttcagtgctg aggcggcggt
     6241 ggcgcgcgcc gccaggcggg ggcgaaggca ctgtccgcgg tgctgaagct ggcagtgcgc
     6301 acgcgcctcg ccgcatcctg tttcccctcc ccctctctga taggggatgc gcaatttggg
     6361 gaatgggggt tgggtgcttg tccagtgggt cggggtcggt cgtcaattta aatactgaaa
     6421 agggcgaatt ccagcacact ggagctcgga tcttcctaga gcatggctac gtagataagt
     6481 agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct
     6541 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     6601 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
//
