LOCUS       sequence-409162-        6491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for glutamate, third generation, variant 857 in
            NGR backbone. iGluSnFR3.v857.NGR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6491)
  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 6491)
  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.v857.IgK-NGR
FEATURES             Location/Qualifiers
     source          1..6491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     protein_bind    137..157
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        158..605
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          698..830
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     protein_bind    830..854
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(855..888)
                     /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)."
     protein_bind    complement(921..954)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1090..1119)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1584..1605
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1708..1729)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2698..2757)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2754..2763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2770..2803
                     /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)."
     protein_bind    2836..2869
                     /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(2870..2894)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2904..3492
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2957..2977)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3375..3386)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3522..3539)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3528..3735
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3743..3883
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3743..3872
                     /label=AAV2 ITR
     rep_origin      3958..4413
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4045..4064)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4255..4276
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4430..4449)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4549..4571
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4609..4627)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4695..4799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4800..4868,4869..5660)
                     /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(5018..5037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5831..6419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6320..6339
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6481..6491,1..130)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tctagacaac tttgtataga aaagttgagt gcaagtgggt tttaggacca
      181 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
      241 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
      301 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
      361 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
      421 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
      481 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
      541 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
      601 cgcagctgtg ctcctgggca ccgcgcagtc cgcccccgcg gctcctggcc agaccacccc
      661 taggaccccc tgccccaagt cgcagccgtt ggatcaggta agtatcaagg ttacaagaca
      721 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
      781 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag caagtttgta
      841 caaaaaagca ggctataact tcgtatagga tactttatac gaagttatcc tcgagtaccg
      901 gtcacgcgta cgcaagcttg ataacttcgt atagcataca ttatacgaag ttatcctagg
      961 gccaccctaa caggggccca acaccgtcca cagaaccaat gccagtccca gtggagccag
     1021 actacatgca agggcaggaa gggcacctga tccttctgca tctcctgttc cagatgcacc
     1081 actacctgcc agatcctctt ctgagatgag tttttgttcg tcgacctgca gtttcagtgc
     1141 cttgtcattc ggttctttga acagtgcttt catttcgtct gacagttcga aattcatgtt
     1201 cagggggttg ttaaagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     1261 gtcgatgccc tttagctcga tgcggttcac cagggtgtcg ccctctaact tcacctcggc
     1321 gcgggtcttg taggttccgt cgtccttgaa gctgatggtg cgctcctgga cgtagccttc
     1381 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca
     1441 ctgcacgccg tagctcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     1501 ggtgcagatg aacttcaggg ttagcttgcc gttggtggca tcgccctcgc cctcgccgcg
     1561 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     1621 ggtgaacagc tcctcgccct tgctcatgct ccctccggta ccgcccttgt acagctcgtc
     1681 catgccgaga gagttcccgg tggcggtcac gaactccagc aggaccatgt gatcgcgctt
     1741 ctcgttgggg tctttgctca gcacggactg atagctcagg tagtggttgt cgggcagcag
     1801 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc
     1861 gtcctccacg ttgtggtgga tcttgaagtt cgccttgatg ccgttcttct gcttgtcggc
     1921 ggtgatatag accgtgtggt tcaccagaat tggatttttg aaccacttat caaaccattt
     1981 ttccgcttca ccggaggtct gcacctgagc gatggtgtca tccatcagct ttttgaactg
     2041 cggatcatct ttacgcaaca tacaaccgta ggcctcctga gactgcggct tgccgacgat
     2101 ttcccagtcg tctggtttct tcgctttcgc acgttcaccg gccagcagcg agtcatccat
     2161 cataaaggca acggcacgac cgctttccag ggtgcggaaa gagtcaccgt gatctttggc
     2221 gctgatgatg cgcatattca ttttttgctc ttcattcagt ttgttgagca aaacttcaga
     2281 ggtagtgccg gaagtgacga ctacggcttt gtctttcagg ttggcaaaat ctttgatatc
     2341 gccacccttt ttggtcaaca ggcgcgtacc gaccacgaaa atagtgtcag agaaagccgc
     2401 ctgtttttgg cgttcgacgt tgttggtggt agaaccacat tcaaaatcgt aagtgccgtt
     2461 ttgcagtagt ggaatacggt tttgtgaagt aatcggaatc agttttacct gcaagtccgg
     2521 tttgttgagt ttctttttca ctgcttcaac aatggcgttg gagtattcct gcgagtaacc
     2581 caccactttg tgctgactgt cgtaataaga gaaaggcact gaagagtcac ggtgaccgac
     2641 gacaatcaca ccgtttttgg cgattttgtc cagcgtgctg cctgcggcgg atccgtcacc
     2701 agtggaacct ggaacccaga gcagcagtac ccatagcagg agtgtgtctg tctccatggt
     2761 ggctctagaa taacttcgta taaagtatcc tatacgaagt tataggtacc tgagctcaac
     2821 atcgatagca ctagtataac ttcgtataat gtatgctata cgaagttata cccagctttc
     2881 ttgtacaaag tgggaattcc gataatcaac ctctggatta caaaatttgt gaaagattga
     2941 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3001 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3061 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3121 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3181 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3241 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     3301 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     3361 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3421 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3481 ccgcctcccc gcatcgggaa ttcctagagc tcgctgatca gcctcgactg tgccttctag
     3541 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3601 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3661 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag
     3721 caggcatgct ggggagggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3781 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3841 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     3901 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3961 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4021 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4081 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4141 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4201 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4261 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     4321 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4381 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4441 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4501 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4561 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4621 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4681 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4741 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4801 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4861 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4921 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4981 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5041 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5101 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5161 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5221 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5281 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5341 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5401 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5461 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc
     5521 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5581 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5641 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5701 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5761 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5821 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5881 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5941 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6001 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6061 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6121 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6181 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6241 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6301 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6361 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6421 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6481 cctgcaggca g
//
