LOCUS       sequence-345963-        6625 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 6625)
  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 6625)
  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.GPI.codonopt
FEATURES             Location/Qualifiers
     source          1..6625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     254..273
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   590..719
                     /label=AAV2 ITR
     polyA_signal    881..1002
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(918..937)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     972..991
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    1079..1667
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1185..1196
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1594..1614
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1735..1768
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1839..1872
                     /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      1891..1900
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1897..1956
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             3535..3564
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(3691..3724)
                     /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(3787..3820)
                     /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(3839..3858)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          3841..3973
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   4572..4701
                     /label=AAV2 ITR
     rep_origin      4773..5228
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4860..4879)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5070..5091
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5254..5358
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5359..5427,5428..6219)
                     /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(5577..5596)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6390..6625,1..353)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
       61 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      121 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      181 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      241 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      301 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      361 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      421 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      481 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      541 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tgcgcgctcg
      601 ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc
      661 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt
      721 gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta ggaagatcct
      781 tatcgatttt accacatttg tagaggtttt acttgcttta aaaaacctcc cacatctccc
      841 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
      901 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
      961 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gctcgaagcg
     1021 gccggccgcc cgggcgacct agatgcattc gacggccgcc cgggtcgacg gtatcgatgc
     1081 ggggaggcgg cccaaaggga gatccgactc gtctgagggc gaaggcgaag acgcggaaga
     1141 ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg ctggattgag ggccgaaggg
     1201 acgtagcaga aggacgtccc gcgcagaatc caggtggcaa cacaggcgag cagccaagga
     1261 aaggacgatg atttccccga caacaccacg gaattgtcag tgcccaacag ccgagcccct
     1321 gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaatagg gagggggaaa
     1381 gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc cccaaccagt gggggttgcg
     1441 tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa cgggccacaa ctcctcataa
     1501 agagacagca accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata
     1561 gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta
     1621 agaataccag tcaatctttc acaaattttg taatccagag gttgattctt aagagcttcg
     1681 agtggatcat ctgcaaattc gcccttttca gtgtttcgaa cctcttcgag ggacataact
     1741 tcgtataatg tatgctatac gaagttatgg gtcgatggtg agatctggac tagagggtcg
     1801 atggtgatgc ttggatcaaa ataggaagaa tcccaatgat aacttcgtat aaagtatcct
     1861 atacgaagtt atactagtca tatgggatcc gccaccatgg agacagacac actcctgcta
     1921 tgggtactgc tgctctgggt tccaggttcc actggtgacg gaagcgctgc aggatcaaca
     1981 ctggacaaaa ttgctaaaaa tggtgtcata gtagttggac acagagactc ttccgtacca
     2041 ttttcttatt acgatagtca gcataaggtg gtcggatatt cacaagaata cagcaatgcc
     2101 atcgtggaag ctgtgaagaa gaaactgaac aaaccagatc ttcaagttaa gttgattcca
     2161 attacttccc agaatcggat acccctcctg caaaacggga cttatgattt cgagtgcggt
     2221 agtactacaa acaacgtcga acgacaaaag caagcagctt tttctgacac tatctttgtt
     2281 gtgggaacca gactgctcac caagaaggga ggggatatta aagactttgc caatctgaag
     2341 gataaggcag tggtggtgac tagtggaaca acttcagagg ttttgctgaa caaactcaat
     2401 gaggaacaaa aaatgaacat gcgcattatt agcgcaaaag atcatggcga ttccttcaga
     2461 acattggaaa gtgggcgcgc tgttgccttc atgatggatg acagcctgct cgccggagaa
     2521 cgcgccaagg caaagaaacc agatgattgg gagatcgtcg gcaagccaca aagccaggag
     2581 gcttatgggt gcatgctcag gaaagatgac ccccaattta agaagctgat ggatgacact
     2641 atagcacaag tccaaacctc cggagaagct gagaagtggt tcgacaaatg gtttaaaaac
     2701 cccatcctgg tcaaccatac cgtctatatc acagccgata agcagaaaaa tggaattaaa
     2761 gccaatttta agatccacca taatgtagaa gatgggagcg tgcagttggc agaccactac
     2821 cagcaaaata cacctatcgg ggatggtcca gtgctgctgc ctgacaacca ctatctctca
     2881 tatcaatctg tgttgtcaaa agatccaaat gaaaaaaggg accatatggt gctgctcgaa
     2941 tttgtaacag ccacagggaa ctcactgggc atggacgagc tctacaaggg agggactggc
     3001 ggctcaatgt ccaaagggga agaattgttt accggagtcg tgcctatcct tgtcgaactg
     3061 gatggagatg ttaatgggca caaattctcc gtacgcggcg agggcgaagg ggacgccact
     3121 aatgggaagc tgactcttaa atttatctgc acaactggta aattgccagt gccctggcct
     3181 acccttgtca ccaccttgag ctacggcgtc cagtgtttcg caaggtatcc agaccatatg
     3241 aagcaacatg acttttttaa atccgctatg cccgagggtt acgtacaaga gagaaccatc
     3301 agtttcaaag atgatggtac ttacaaaact cgcgcagaag tcaagctcga aggagacacc
     3361 cttgtaaatc gaatagagtt gaagggcatt gattttaagg aagacggtaa tatcctcggc
     3421 cacaaactcg agtataattt taacaacccc ttgaatatga actttgaact ctcagatgaa
     3481 atgaaagccc tgtttaaaga gccaaacgac aaagcactta aactgcaggt cgacgaacaa
     3541 aaactcatct cagaagagga tctgtcagat aacgactacg gtaaagatgg gttggctggg
     3601 gcttcaggag gcagacgaaa tggcgctata actgtgctca gtttcataac gttctacgtg
     3661 gcagcattta tggtattgtt gtagaagctt ataacttcgt atagcataca ttatacgaag
     3721 ttatgtgtcg atggtgatgc ttggcaattc gggtcgatgg tgaagcattg gtcttcctat
     3781 tttgatataa cttcgtatag gatactttat acgaagttat ggggggcgcg ccatcatcac
     3841 ctgtggagag aaaggcaaag tggatgtcag taagaccaat aggtgcctat cagaaacgca
     3901 agagtcttct ctgtctcgac aagcccagtt tctattggtc tccttaaacc tgtcttgtaa
     3961 ccttgatact tacctgatcc aacggctgcg acttggggca gggggtccta ggggtggtct
     4021 ggccaggagc cgcgggggcg gactgcgcgg tgcccaggag cacagctgcg ctctcaggca
     4081 cgacacgact cctccgctgc ccaccgcaga ctgaggcagc gctgagtcgc cggcgccgca
     4141 gcgcagatgg tcgcgcccgt gcccccctat ctcgcgcctc gcgtggtgcg gtccggctgg
     4201 gccggcggcg gcgcggacgc gaccaaggtg gccgggaagg ggagtttgcg ggggaccggc
     4261 gagtgacgtc agcgcgcctt cagtgctgag gcggcggtgg cgcgcgccgc caggcggggg
     4321 cgaaggcact gtccgcggtg ctgaagctgg cagtgcgcac gcgcctcgcc gcatcctgtt
     4381 tcccctcccc ctctctgata ggggatgcgc aatttgggga atgggggttg ggtgcttgtc
     4441 cagtgggtcg gggtcggtcg tcaagcatgc tttaaatact gaaaagggcc aattccagca
     4501 cactggagct cggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat
     4561 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     4621 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     4681 agtgagcgag cgagcgcgca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4741 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     4801 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4861 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4921 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4981 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5041 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5101 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5161 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     5221 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5281 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5341 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5401 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5461 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5521 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5581 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5641 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5701 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5761 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5821 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5881 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5941 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6001 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6061 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6121 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6181 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6241 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6301 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6361 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6421 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6481 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6541 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6601 gctctgctaa tcctgttacc agtgg
//
