LOCUS       sequence-409161-        6069 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 6069)
  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 6069)
  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.iGluSnFR3.v857.IgK-NGR
FEATURES             Location/Qualifiers
     source          1..6069
                     /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        179..626
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    626..650
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      651..660
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     657..716
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1685..1706
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1809..1830)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2295..2324
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(2448..2472)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2482..3070
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2535..2555)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2953..2964)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3100..3117)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3106..3313
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3321..3461
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3321..3450
                     /label=AAV2 ITR
     rep_origin      3536..3991
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3623..3642)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3833..3854
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4008..4027)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4127..4149
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4187..4205)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4273..4377
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4378..4446,4447..5238)
                     /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(4596..4615)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5409..5997
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5898..5917
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6059..6069,1..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 aaagttgctg cagagggccc tgcgtatgag
      181 tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc
      241 ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga
      301 gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga
      361 cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc
      421 tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg
      481 ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg
      541 accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag
      601 gagtcgtgtc gtgcctgaga gcgcagcaag tttgtacaaa aaagcaggct gccaccatgg
      661 agacagacac actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgacg
      721 gatccgccgc aggcagcacg ctggacaaaa tcgccaaaaa cggtgtgatt gtcgtcggtc
      781 accgtgactc ttcagtgcct ttctcttatt acgacagtca gcacaaagtg gtgggttact
      841 cgcaggaata ctccaacgcc attgttgaag cagtgaaaaa gaaactcaac aaaccggact
      901 tgcaggtaaa actgattccg attacttcac aaaaccgtat tccactactg caaaacggca
      961 cttacgattt tgaatgtggt tctaccacca acaacgtcga acgccaaaaa caggcggctt
     1021 tctctgacac tattttcgtg gtcggtacgc gcctgttgac caaaaagggt ggcgatatca
     1081 aagattttgc caacctgaaa gacaaagccg tagtcgtcac ttccggcact acctctgaag
     1141 ttttgctcaa caaactgaat gaagagcaaa aaatgaatat gcgcatcatc agcgccaaag
     1201 atcacggtga ctctttccgc accctggaaa gcggtcgtgc cgttgccttt atgatggatg
     1261 actcgctgct ggccggtgaa cgtgcgaaag cgaagaaacc agacgactgg gaaatcgtcg
     1321 gcaagccgca gtctcaggag gcctacggtt gtatgttgcg taaagatgat ccgcagttca
     1381 aaaagctgat ggatgacacc atcgctcagg tgcagacctc cggtgaagcg gaaaaatggt
     1441 ttgataagtg gttcaaaaat ccaattctgg tgaaccacac ggtctatatc accgccgaca
     1501 agcagaagaa cggcatcaag gcgaacttca agatccacca caacgtggag gacggcagcg
     1561 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1621 ccgacaacca ctacctgagc tatcagtccg tgctgagcaa agaccccaac gagaagcgcg
     1681 atcacatggt cctgctggag ttcgtgaccg ccaccgggaa ctctctcggc atggacgagc
     1741 tgtacaaggg cggtaccgga gggagcatga gcaagggcga ggagctgttc accggggtgg
     1801 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgcgcggcg
     1861 agggcgaggg cgatgccacc aacggcaagc taaccctgaa gttcatctgc accaccggca
     1921 agctgcccgt gccctggccc accctcgtga ccaccctgag ctacggcgtg cagtgcttcg
     1981 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2041 acgtccagga gcgcaccatc agcttcaagg acgacggaac ctacaagacc cgcgccgagg
     2101 tgaagttaga gggcgacacc ctggtgaacc gcatcgagct aaagggcatc gacttcaagg
     2161 aggacggcaa catcctgggg cacaagctgg agtacaactt taacaacccc ctgaacatga
     2221 atttcgaact gtcagacgaa atgaaagcac tgttcaaaga accgaatgac aaggcactga
     2281 aactgcaggt cgacgaacaa aaactcatct cagaagagga tctggcaggt agtggtgcat
     2341 ctggaacagg agatgcagaa ggatcaggtg cccttcctgc ccttgcatgt agtctggctc
     2401 cactgggact ggcattggtt ctgtggacgg tgttgggccc ctgttagacc cagctttctt
     2461 gtacaaagtg ggaattccga taatcaacct ctggattaca aaatttgtga aagattgact
     2521 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2581 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2641 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2701 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2761 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2821 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     2881 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     2941 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3001 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3061 gcctccccgc atcgggaatt cctagagctc gctgatcagc ctcgactgtg ccttctagtt
     3121 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3181 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3241 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca
     3301 ggcatgctgg ggagggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     3361 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3421 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     3481 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     3541 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3601 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3661 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3721 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3781 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3841 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     3901 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3961 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4021 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4081 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4141 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4201 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4261 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4321 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4381 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4441 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4501 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4561 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4621 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4681 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4741 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4801 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4861 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4921 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4981 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5041 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc
     5101 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5161 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5221 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5281 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5341 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5401 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5461 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5521 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5581 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5641 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5701 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5761 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5821 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5881 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5941 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6001 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc
     6061 tgcaggcag
//
