LOCUS       sequence-405486-        6525 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 6525)
  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 6525)
  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.GPI
FEATURES             Location/Qualifiers
     source          1..6525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      42..497
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(129..148)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     339..360
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        523..627
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(628..696,697..1488)
                     /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(846..865)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1659..2247
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2148..2167
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2484..2613
                     /label=AAV2 ITR
     polyA_signal    2775..2896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2812..2831)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2866..2885
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2925..2941
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2926..2942
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2945..3533
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3051..3062
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3460..3480
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3630..3677
                     /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      3713..3722
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3719..3778
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             5357..5386
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(5614..5661)
                     /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(5681..5700)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          5683..5815
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(5837..6284)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   6366..6495
                     /label=AAV2 ITR
ORIGIN
        1 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
       61 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      121 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      181 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      241 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      301 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      361 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      421 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
      481 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
      541 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
      601 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
      661 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
      721 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
      781 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
      841 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
      901 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
      961 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1021 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1081 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1141 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1201 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1261 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1321 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1381 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1441 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     1501 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1561 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1621 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1681 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1741 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1801 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1861 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1921 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1981 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2041 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2101 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2161 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2221 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2281 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2341 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2401 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     2461 cgttggccga ttcattaatg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     2521 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     2581 agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac ccgccatgct
     2641 acttatctac gtagccatgc tctaggaaga tccttatcga ttttaccaca tttgtagagg
     2701 ttttacttgc tttaaaaaac ctcccacatc tccccctgaa cctgaaacat aaaatgaatg
     2761 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2821 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2881 tcatcaatgt atcttatcat gtctgctcga agcggccgcc cgggtcgagg tcgacggtat
     2941 cgatgcgggg aggcggccca aagggagatc cgactcgtct gagggcgaag gcgaagacgc
     3001 ggaagaggcc gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg attgagggcc
     3061 gaagggacgt agcagaagga cgtcccgcgc agaatccagg tggcaacaca ggcgagcagc
     3121 caaggaaagg acgatgattt ccccgacaac accacggaat tgtcagtgcc caacagccga
     3181 gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aatagggagg
     3241 gggaaagcga aagtcccgga aaggagctga caggtggtgg caatgcccca accagtgggg
     3301 gttgcgtcag caaacacagt gcacaccacg ccacgttgcc tgacaacggg ccacaactcc
     3361 tcataaagag acagcaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa
     3421 gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca
     3481 tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg attatcgata
     3541 agcttcctgc agggaagttc ctattccgaa gttcctattc tgaactaagt ataggaactt
     3601 cttggatcaa aataggagcg atcgcaatgg aagttcctat tccgaagttc ctattctcta
     3661 gaaagtatag gaacttcgtt taaactggcc ggccactagt agatctgaat tcgccaccat
     3721 ggagacagac acactcctgc tatgggtact gctgctctgg gttccaggtt ccactggtga
     3781 cggaagcgct gcaggatcaa cactggacaa aattgctaaa aatggtgtca tagtagttgg
     3841 acacagagac tcttccgtac cattttctta ttacgatagt cagcataagg tggtcggata
     3901 ttcacaagaa tacagcaatg ccatcgtgga agctgtgaag aagaaactga acaaaccaga
     3961 tcttcaagtt aagttgattc caattacttc ccagaatcgg atacccctcc tgcaaaacgg
     4021 gacttatgat ttcgagtgcg gtagtactac aaacaacgtc gaacgacaaa agcaagcagc
     4081 tttttctgac actatctttg ttgtgggaac cagactgctc accaagaagg gaggggatat
     4141 taaagacttt gccaatctga aggataaggc agtggtggtg actagtggaa caacttcaga
     4201 ggttttgctg aacaaactca atgaggaaca aaaaatgaac atgcgcatta ttagcgcaaa
     4261 agatcatggc gattccttca gaacattgga aagtgggcgc gctgttgcct tcatgatgga
     4321 tgacagcctg ctcgccggag aacgcgccaa ggcaaagaaa ccagatgatt gggagatcgt
     4381 cggcaagcca caaagccagg aggcttatgg gtgcatgctc aggaaagatg acccccaatt
     4441 taagaagctg atggatgaca ctatagcaca agtccaaacc tccggagaag ctgagaagtg
     4501 gttcgacaaa tggtttaaaa accccatcct ggtcaaccat accgtctata tcacagccga
     4561 taagcagaaa aatggaatta aagccaattt taagatccac cataatgtag aagatgggag
     4621 cgtgcagttg gcagaccact accagcaaaa tacacctatc ggggatggtc cagtgctgct
     4681 gcctgacaac cactatctct catatcaatc tgtgttgtca aaagatccaa atgaaaaaag
     4741 ggaccatatg gtgctgctcg aatttgtaac agccacaggg aactcactgg gcatggacga
     4801 gctctacaag ggagggactg gcggctcaat gtccaaaggg gaagaattgt ttaccggagt
     4861 cgtgcctatc cttgtcgaac tggatggaga tgttaatggg cacaaattct ccgtacgcgg
     4921 cgagggcgaa ggggacgcca ctaatgggaa gctgactctt aaatttatct gcacaactgg
     4981 taaattgcca gtgccctggc ctacccttgt caccaccttg agctacggcg tccagtgttt
     5041 cgcaaggtat ccagaccata tgaagcaaca tgactttttt aaatccgcta tgcccgaggg
     5101 ttacgtacaa gagagaacca tcagtttcaa agatgatggt acttacaaaa ctcgcgcaga
     5161 agtcaagctc gaaggagaca cccttgtaaa tcgaatagag ttgaagggca ttgattttaa
     5221 ggaagacggt aatatcctcg gccacaaact cgagtataat tttaacaacc ccttgaatat
     5281 gaactttgaa ctctcagatg aaatgaaagc cctgtttaaa gagccaaacg acaaagcact
     5341 taaactgcag gtcgacgaac aaaaactcat ctcagaagag gatctgtcag ataacgacta
     5401 cggtaaagat gggttggctg gggcttcagg aggcagacga aatggcgcta taactgtgct
     5461 cagtttcata acgttctacg tggcagcatt tatggtattg ttgtagtaaa ccggtacgcg
     5521 tttaattaag tttaaacgaa gttcctatac ttagttcaga ataggaactt cggaatagga
     5581 acttcagagc gctttggccg ctccggccaa agggaagttc ctatactttc tagagaatag
     5641 gaacttcgga ataggaactt cggcgcgccg gatccttatc acctgtggag agaaaggcaa
     5701 agtggatgtc agtaagacca ataggtgcct atcagaaacg caagagtctt ctctgtctcg
     5761 acaagcccag tttctattgg tctccttaaa cctgtcttgt aaccttgata cttacctgct
     5821 agcagcttga attcgactgc gctctcaggc acgacacgac tcctccgctg cccaccgcag
     5881 actgaggcag cgctgagtcg ccggcgccgc agcgcagatg gtcgcgcccg tgccccccta
     5941 tctcgcgcct cgcgtggtgc ggtccggctg ggccggcggc ggcgcggacg cgaccaaggt
     6001 ggccgggaag gggagtttgc gggggaccgg cgagtgacgt cagcgcgcct tcagtgctga
     6061 ggcggcggtg gcgcgcgccg ccaggcgggg gcgaaggcac tgtccgcggt gctgaagctg
     6121 gcagtgcgca cgcgcctcgc cgcatcctgt ttcccctccc cctctctgat aggggatgcg
     6181 caatttgggg aatgggggtt gggtgcttgt ccagtgggtc ggggtcggtc gtcaggtagg
     6241 cacccccacc ccgcctcatc ctggtcctaa aacccacttg cactcatacg cagggccctc
     6301 tgcagagatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
     6361 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     6421 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     6481 cgagcgagcg cgcagctggc gtaatagcga agaggcccgc accga
//
