LOCUS       sequence-204217-        6905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tight affinity yellow glutamate sensor ** A newer version of this 
            sensor is available. Please see iGluSnFR3 plasmids at 
            https://www.addgene.org/browse/article/28220233/ **.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6905)
  AUTHORS   Marvin JS, Scholl B, Wilson DE, Podgorski K, Kazemipour A, Muller 
            JA, Schoch S, Quiroz FJU, Rebola N, Bao H, Little JP, Tkachuk AN, 
            Cai E, Hantman AW, Wang SS, DePiero VJ, Borghuis BG, Chapman ER, 
            Dietrich D, DiGregorio DA, Fitzpatrick D, Looger LL
  TITLE     Stability, affinity, and chromatic variants of the glutamate sensor 
            iGluSnFR.
  JOURNAL   Nat Methods. 2018 Nov;15(11):936-939. doi: 
            10.1038/s41592-018-0171-3. Epub 2018 Oct 30.
  PUBMED    30377363
REFERENCE   2  (bases 1 to 6905)
  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"; doi: 
            "10.1038/s41592-018-0171-3"; journalName: "Nat Methods"; date: 
            "2018-11"; volume: "15"; issue: "11"; pages: "936-939"
COMMENT     Sequence Label: pAAV.CAG-FLEX.SF-Venus-iGluSnFR.A184S
FEATURES             Location/Qualifiers
     source          1..6905
                     /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    1171..1204
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1275..1308
                     /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      1333..1342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1339..1398
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     2367..2388
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2491..2512)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2977..3006
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3010..3156
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(3184..3217)
                     /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(3280..3313)
                     /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(3348..3367)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3429..3451)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3730..4007)
                     /label=chicken beta-actin promoter
     repeat_region   4318..4447
                     /label=AAV2 ITR
     rep_origin      4519..4974
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4606..4625)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4816..4837
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5000..5104
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5105..5173,5174..5965)
                     /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(5323..5342)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6136..6724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6625..6644
                     /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 tctcgagtgg atcatttgca gaattggccc
     1141 ttttcagtgt ttcgaacctc ttcgagggac ataacttcgt ataatgtatg ctatacgaag
     1201 ttatgggtcg atggtgagat ttggactaga gggtcgatgg tgatgcttgg atcaaaatag
     1261 gaagaatccc aatgataact tcgtataaag tatcctatac gaagttatac tagtggtacc
     1321 accggtgata tcgccaccat ggagacagac acactcctgc tatgggtact gctgctctgg
     1381 gttccaggtt ccactggtga cagatctgcc gcaggcagca cgctggacaa aatcgccaaa
     1441 aacggtgtga ttgtcgtcgg tcaccgtgaa tcttcagtgc ctttctctta ttacgacaat
     1501 cagcaaaaag tggtgggtta ctcgcaggat tactccaacg ccattgttga agcagtgaaa
     1561 aagaaactca acaaaccgga cttgcaggta aaactgattc cgattacctc acaaaaccgt
     1621 attccactgc tgcaaaacgg cactttcgat tttgaatgtg gttctaccac caacaacgtc
     1681 gaacgccaaa aacaggcggc tttctctgac actattttcg tggtcggtac gcgcctgttg
     1741 accaaaaagg gtggcgatat caaagatttt gccaacctga aagacaaagc cgtagtcgtc
     1801 acttccggca ctacctctga agttttgctc aacaaactga atgaagagca aaaaatgaat
     1861 atgcgcatca tcagcgccaa agatcacggt gactctttcc gcaccctgga aagcggtcgt
     1921 gccgttgcct ttatgatgga tgacagcctg ctggccggtg aacgtgcgaa agcgaagaaa
     1981 ccagacaact gggaaatcgt cggcaagccg cagtctcagg aggcctacgg ttgtatgttg
     2041 cgtaaagatg atccgcagtt caaaaagctg atggatgaca ccatcgctca ggtgcagacc
     2101 tccggtgaag cggaaaaatg gtttgataag tggttcaaaa atccaattct ggtgagccac
     2161 aacgtctata tcaccgccga caagcagaag aacggcatca aggcgaactt caagatccgc
     2221 cacaacgtgg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2281 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctatcagtc cgtgctgagc
     2341 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2401 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat gagcaagggc
     2461 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2521 cacaagttca gcgtgcgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg
     2581 aagctcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2641 ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2701 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcagcttcaa ggacgacggc
     2761 acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2821 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2881 tttaacaacc ccctgaacat gaatttcgaa ctgtcagacg aaatgaaagc actgttcaaa
     2941 gaaccgaatg acaaggcact gaaactgcag gtcgacgaac aaaaactcat ctcagaagag
     3001 gatctgaatg ctgtgggcca ggacacgcag gaggtcatcg tggtgccaca ctccttgccc
     3061 tttaaggtgg tggtgatctc agccatcctg gccctggtgg tgctcaccat catctccctt
     3121 atcatcctca tcatgctttg gcagaagaag ccacgttagg atatcggatc cgctagcact
     3181 agtataactt cgtatagcat acattatacg aagttatggg tcgatggtga tgcttggcaa
     3241 ttcgggtcga tggtgaagca ttggtcttcc tattttgata taacttcgta taggatactt
     3301 tatacgaagt tatggggatc caattctttg ccaaaatgat gagacagcac aataaccagc
     3361 acgttgccca ggagctgtag gaaaaagaag aaggcatgaa catggttagc agaggctcta
     3421 gagccgccgg tcacacgcca gaagccgaac cccgccctgc cccgtccccc ccgaaggcag
     3481 ccgtcccccc gcggacagcc ccgagccgct ccccccgcac aaagggccct cccggagccc
     3541 ctcaaggctt tcacgcagcc acagaaaaga aacaagccgt cattaaacca agcgctaatt
     3601 acagcccgga ggagaagggc cgtcccgccc gctcacctgt gggagtaacg cggtcagtca
     3661 gagccggggc gggcggcgcg aggcggcggc ggagcggggc acggggcgaa ggcagcgcgc
     3721 agcgactccc gcccgccgcg cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc
     3781 cataaaagga aactttcgga gcgcgccgct ctgattggct gccgccgcac ctctccgcct
     3841 cgccscgccc csccccycsc cccgccccgc cccgcctggc gcgcgccccc cccccccccc
     3901 cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg ggggtgggga
     3961 ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca tggtaatagc
     4021 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     4081 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     4141 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     4201 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatggtac
     4261 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     4321 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     4381 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     4441 cgcgcagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4501 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4561 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4621 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4681 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4741 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4801 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4861 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4921 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     4981 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5041 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5101 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5161 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5221 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5281 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5341 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5401 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5461 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5521 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5581 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5641 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5701 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5761 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5821 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5881 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5941 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6001 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6061 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6181 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6241 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6301 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6361 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6421 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6481 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6541 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6601 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6661 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6721 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6781 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6841 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6901 ggaag
//
