LOCUS       sequence-405480-        6847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for glutamate, third generation, variant 857 in
            PDGFR backbone. iGluSnFR3.v857.PDGFR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6847)
  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 6847)
  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.CAG.FLEX.iGluSnFR3.v857.PDGFR
FEATURES             Location/Qualifiers
     source          1..6847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1133..1166
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1237..1270
                     /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      1278..1287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1284..1343
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2928..2957
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2961..3107
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(3123..3156)
                     /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(3219..3252)
                     /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(3290..3309)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3371..3393)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3672..3949)
                     /label=chicken beta-actin promoter
     repeat_region   4260..4389
                     /label=AAV2 ITR
     rep_origin      4461..4916
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4548..4567)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4758..4779
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4942..5046
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5047..5115,5116..5907)
                     /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(5265..5284)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6078..6666
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6567..6586
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct aattcgaacc tcttcgaggg acataacttc
     1141 gtataatgta tgctatacga agttatgggt cgatggtgag atctggacta gagggtcgat
     1201 ggtgatgctt ggatcaaaat aggaagaatc ccaatgataa cttcgtataa agtatcctat
     1261 acgaagttat ggatcccgcc accatggaga cagacacact cctgctatgg gtactgctgc
     1321 tctgggttcc aggttccact ggtgacagat ctgggtccgc cgccggaagt actcttgata
     1381 agatcgctaa aaatggtgta attgttgtcg gccaccgaga ctcttctgtg ccattcagct
     1441 attatgattc tcaacacaaa gtggtgggct attcacaaga gtattctaac gccattgttg
     1501 aagcagttaa gaagaagttg aataaaccag accttcaagt aaaactcatt cctataacat
     1561 cccaaaaccg catacctctc ttgcaaaacg gcacttacga tttcgagtgt gggagcacta
     1621 ctaataatgt cgaacgccag aaacaagccg cattctccga caccatcttc gtggtgggca
     1681 ctcgactgtt gacaaagaaa ggcggtgata ttaaggattt cgcaaatctg aaggataagg
     1741 cagtcgtggt caccagcggg accacctcag aagtactcct taataaactc aatgaagaac
     1801 aaaagatgaa tatgcgaatc atctctgcta aggaccacgg tgactctttt cgcacattgg
     1861 aaagtgggcg cgccgtagca tttatgatgg acgactctct gctcgctggc gagcgcgcca
     1921 aagccaaaaa accagacgac tgggaaattg taggtaaacc tcaaagccaa gaggcatatg
     1981 gctgcatgtt gagaaaagat gacccccaat tcaagaaact gatggacgac actatagctc
     2041 aggtccagac ttcaggggaa gctgagaagt ggtttgataa atggttcaag aacccaatct
     2101 tggtaaatca taccgtttat attaccgctg acaaacaaaa gaatgggata aaggcaaatt
     2161 tcaagattca ccataacgta gaggatggaa gcgtgcagtt ggcagaccat tatcaacaga
     2221 atacccctat tggcgacgga cccgtgctcc tccctgacaa ccactacttg tcctatcaaa
     2281 gtgttctctc aaaggacccc aatgagaagc gagaccacat ggtacttctc gaattcgtta
     2341 ccgccaccgg gaattcactt ggcatggatg aattgtataa ggggggtact ggcggcagca
     2401 tgtccaaggg tgaggagctt tttaccggag tagtaccaat tctcgttgaa cttgacggag
     2461 acgtcaacgg gcacaagttt agcgtacgag gtgaggggga gggtgatgca accaacggta
     2521 aacttactct caaattcata tgcactaccg gaaagctgcc agtaccttgg ccaacactgg
     2581 tcactaccct ctcctatggc gttcaatgtt ttgctcgata ccctgaccac atgaaacagc
     2641 atgacttctt taaatcagcc atgcctgaag ggtatgtcca ggagcggact atatcattta
     2701 aagacgatgg aacctataag actcgggctg aagtgaagct cgagggtgac acattggtta
     2761 atcggatcga gcttaagggc atagatttta aagaagatgg taatattctt ggccacaaat
     2821 tggagtacaa ttttaataat cctttgaaca tgaactttga gctctccgat gaaatgaagg
     2881 cactctttaa agagccaaac gacaaagctc tgaaactgca ggtcgacgaa caaaaactca
     2941 tctcagaaga ggatctgaat gctgtgggcc aggacacgca ggaggtcatc gtggtgccac
     3001 actccttgcc ctttaaggtg gtggtgatct cagccatcct ggccctggtg gtgctcacca
     3061 tcatctccct tatcatcctc atcatgcttt ggcagaagaa gccacgttag aagcttgcta
     3121 gtataacttc gtatagcata cattatacga agttatgggt cgatggtgat gcttggcaat
     3181 tcgggtcgat ggtgaagcat tggtcttcct attttgatat aacttcgtat aggatacttt
     3241 atacgaagtt atggggatga tccaattctt tgccaaaatg atgagacagc acaataacca
     3301 gcacgttgcc caggagctgt aggaaaaaga agaaggcatg aacatggtta gcagaggctc
     3361 tagagccgcc ggtcacacgc cagaagccga accccgccct gccccgtccc ccccgaaggc
     3421 agccgtcccc ccgcggacag ccccgagccg ctccccccgc acaaagggcc ctcccggagc
     3481 ccctcaaggc tttcacgcag ccacagaaaa gaaacaagcc gtcattaaac caagcgctaa
     3541 ttacagcccg gaggagaagg gccgtcccgc ccgctcacct gtgggagtaa cgcggtcagt
     3601 cagagccggg gcgggcggcg cgaggcggcg gcggagcggg gcacggggcg aaggcagcgc
     3661 gcagcgactc ccgcccgccg cgcgcttcgc tttttatagg gccgccgccg ccgccgcctc
     3721 gccataaaag gaaactttcg gagcgcgccg ctctgattgg ctgccgccgc acctctccgc
     3781 ctcgccccgc cccgcccctc gccccgcccc gccccgcctg gcgcgcgccc cccccccccc
     3841 cccgccccca tcgctgcaca aaataattaa aaaataaata aatacaaaat tgggggtggg
     3901 gagggggggg agatggggag agtgaagcag aacgtggggc tcacctcgac catggtaata
     3961 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     4021 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     4081 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     4141 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggt
     4201 actcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     4261 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4321 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4381 agcgcgcagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     4441 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4501 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4561 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4621 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4681 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4741 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4801 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4861 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     4921 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4981 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5041 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5101 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5161 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5221 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5281 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5341 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5401 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5461 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5521 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5581 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5641 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5701 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5761 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5821 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5881 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5941 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6001 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6061 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6121 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6181 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6241 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6301 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6361 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6421 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6481 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6541 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6601 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6661 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6721 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6781 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6841 gcggaag
//
