LOCUS       sequence-405478-        6780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for glutamate, third generation, variant 857 in
            SGZ backbone. iGluSnFR3.v857.SGZ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6780)
  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 6780)
  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.FLP-FRT.iGluSnFR3.v857.SGZ
FEATURES             Location/Qualifiers
     source          1..6780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     polyA_signal    2750..2871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2787..2806)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2841..2860
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2900..2916
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2901..2917
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2920..3508
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3026..3037
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3435..3455
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3605..3652
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      3688..3697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3694..3753
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             5332..5361
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5365..5511
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(5844..5891)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(5911..5930)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          5913..6045
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(6067..6514)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   6596..6725
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcctt atcgatttta ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2701 acatctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2761 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2821 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     2881 ctcgaagcgg ccgcccgggt cgaggtcgac ggtatcgatg cggggaggcg gcccaaaggg
     2941 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
     3001 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
     3061 cgcgcagaat ccaggtggca acacaggcga gcagccaagg aaaggacgat gatttccccg
     3121 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     3181 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
     3241 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     3301 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     3361 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     3421 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     3481 cacaaatttt gtaatccaga ggttgattat cgataagctt cctgcaggga agttcctatt
     3541 ccgaagttcc tattctgaac taagtatagg aacttcttgg atcaaaatag gagcgatcgc
     3601 aatggaagtt cctattccga agttcctatt ctctagaaag tataggaact tcgtttaaac
     3661 tggccggcca ctagtagatc tgaattcgcc accatggaga cagacacact cctgctatgg
     3721 gtactgctgc tctgggttcc aggttccact ggtgacggat ccgccgccgg aagtactctt
     3781 gataagatcg ctaaaaatgg tgtaattgtt gtcggccacc gagactcttc tgtgccattc
     3841 agctattatg attctcaaca caaagtggtg ggctattcac aagagtattc taacgccatt
     3901 gttgaagcag ttaagaagaa gttgaataaa ccagaccttc aagtaaaact cattcctata
     3961 acatcccaaa accgcatacc tctcttgcaa aacggcactt acgatttcga gtgtgggagc
     4021 actactaata atgtcgaacg ccagaaacaa gccgcattct ccgacaccat cttcgtggtg
     4081 ggcactcgac tgttgacaaa gaaaggcggt gatattaagg atttcgcaaa tctgaaggat
     4141 aaggcagtcg tggtcaccag cgggaccacc tcagaagtac tccttaataa actcaatgaa
     4201 gaacaaaaga tgaatatgcg aatcatctct gctaaggacc acggtgactc ttttcgcaca
     4261 ttggaaagtg ggcgcgccgt agcatttatg atggacgact ctctgctcgc tggcgagcgc
     4321 gccaaagcca aaaaaccaga cgactgggaa attgtaggta aacctcaaag ccaagaggca
     4381 tatggctgca tgttgagaaa agatgacccc caattcaaga aactgatgga cgacactata
     4441 gctcaggtcc agacttcagg ggaagctgag aagtggtttg ataaatggtt caagaaccca
     4501 atcttggtaa atcataccgt ttatattacc gctgacaaac aaaagaatgg gataaaggca
     4561 aatttcaaga ttcaccataa cgtagaggat ggaagcgtgc agttggcaga ccattatcaa
     4621 cagaataccc ctattggcga cggacccgtg ctcctccctg acaaccacta cttgtcctat
     4681 caaagtgttc tctcaaagga tcccaatgag aagcgagacc acatggtact tctcgaattc
     4741 gttaccgcca ccgggaattc acttggcatg gatgaattgt ataagggggg tactggcggc
     4801 agcatgtcca agggtgagga gctttttacc ggagtagtac caattctcgt tgaacttgac
     4861 ggagacgtca acgggcacaa gtttagcgta cgaggtgagg gggagggtga tgcaaccaac
     4921 ggtaagctta ctctcaaatt catatgcact accggaaagc tgccagtacc ttggccaaca
     4981 ctggtcacta ccctctccta tggcgttcaa tgttttgctc gataccctga ccacatgaaa
     5041 cagcatgact tctttaaatc agccatgcct gaagggtatg tccaggagcg gactatatca
     5101 tttaaagacg atggaaccta taagactcgg gctgaagtga agctcgaggg tgacacattg
     5161 gttaatcgga tcgagcttaa gggcatagat tttaaagaag atggtaatat tcttggccac
     5221 aaattggagt acaattttaa taatcctttg aacatgaact ttgagctctc cgatgaaatg
     5281 aaggcactct ttaaagagcc aaacgacaaa gctctgaaac tgcaggtcga cgaacaaaaa
     5341 ctcatctcag aagaggatct gaatgctgtg ggccaggaca cgcaggaggt catcgtggtg
     5401 ccacactcct tgccctttaa ggtggtggtg atctcagcca tcctggccct ggtggtgctc
     5461 accatcatct cccttatcat cctcatcatg ctttggcaga agaagccacg tgatagacac
     5521 aagcagttgc gagctacagc acgcgccacc gattatctgc aagctgatgc aataactaga
     5581 atccctgatt atagatacag atatcaacga agggatcgcg atgatgatcg agatacagag
     5641 ccagatcatg atcgggatgc tgatcccgtc ggggtgaaag gtttcaacac tcttccagat
     5701 acagagatag ataatcccca gacgacttca gtgtagtaaa ccggtacgcg tttaattaag
     5761 tttaaacgaa gttcctatac ttagttcaga ataggaactt cggaatagga acttcagagc
     5821 gctttggccg ctccggccaa agggaagttc ctatactttc tagagaatag gaacttcgga
     5881 ataggaactt cggcgcgccg gatccttatc acctgtggag agaaaggcaa agtggatgtc
     5941 agtaagacca ataggtgcct atcagaaacg caagagtctt ctctgtctcg acaagcccag
     6001 tttctattgg tctccttaaa cctgtcttgt aaccttgata cttacctgct agcagcttga
     6061 attcgactgc gctctcaggc acgacacgac tcctccgctg cccaccgcag actgaggcag
     6121 cgctgagtcg ccggcgccgc agcgcagatg gtcgcgcccg tgccccccta tctcgcgcct
     6181 cgcgtggtgc ggtccggctg ggccggcggc ggcgcggacg cgaccaaggt ggccgggaag
     6241 gggagtttgc gggggaccgg cgagtgacgt cagcgcgcct tcagtgctga ggcggcggtg
     6301 gcgcgcgccg ccaggcgggg gcgaaggcac tgtccgcggt gctgaagctg gcagtgcgca
     6361 cgcgcctcgc cgcatcctgt ttcccctccc cctctctgat aggggatgcg caatttgggg
     6421 aatgggggtt gggtgcttgt ccagtgggtc ggggtcggtc gtcaggtagg cacccccacc
     6481 ccgcctcatc ctggtcctaa aacccacttg cactcatacg cagggccctc tgcagagatc
     6541 ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa
     6601 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     6661 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     6721 cgcagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
//
