LOCUS       sequence-190085-        6676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre dependent AAV expression of glutamate sensor from human synapsin
            promoter. ** A newer version of this sensor is available.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 6676)
  AUTHORS   Marvin JS, Borghuis BG, Tian L, Cichon J, Harnett MT, Akerboom J, 
            Gordus A, Renninger SL, Chen TW, Bargmann CI, Orger MB, Schreiter 
            ER, Demb JB, Gan WB, Hires SA, Looger LL
  TITLE     An optimized fluorescent probe for visualizing glutamate 
            neurotransmission.
  JOURNAL   Nat Methods. 2013 Feb;10(2):162-70. doi: 10.1038/nmeth.2333. Epub 
            2013 Jan 13.
  PUBMED    23314171
REFERENCE   2  (bases 1 to 6676)
  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/nmeth.2333"; journalName: "Nat Methods"; date: "2013-02"; 
            volume: "10"; issue: "2"; pages: "162-70"
COMMENT     Sequence Label: pAAV.hSyn.Flex.iGluSnFr.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..133
                     /label=AAV2 ITR
     intron          726..858
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     841..860
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    871..904
                     /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)."
     protein_bind    967..1000
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1028..1174)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1178..1207)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1413..1432
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1672..1693
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1799..1820)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2789..2848)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2845..2854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2879..2912)
                     /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)."
     protein_bind    complement(2983..3016)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3088..3676
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3141..3161)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3559..3570)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3754..3875
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3765..3784)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3819..3838
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4037..4166
                     /label=AAV2 ITR
     rep_origin      complement(4403..4991)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4483..4502)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5162..5953,5954..6022))
                     /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     5785..5804
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6023..6127)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6153..6608)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6290..6311)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6502..6521
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggaaga tccgagctcc agtgtgctgg aattcgccct tttcagtatt taaattgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagctgtgct cctgggcacc gcgcagtccg
      661 cccccgcggc tcctggccag accaccccta ggaccccctg ccccaagtcg cagccgttgg
      721 atcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt
      781 cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact
      841 ttgcctttct ctccacaggt gatgatcccc ataacttcgt ataaagtatc ctatacgaag
      901 ttatatcaaa ataggaagac caatgcttca ccatcgaccc gaattgccaa gcatcaccat
      961 cgacacataa cttcgtataa tgtatgctat acgaagttat actagtgcta gcggatccga
     1021 tatcctaacg tggcttcttc tgccaaagca tgatgaggat gataagggag atgatggtga
     1081 gcaccaccag ggccaggatg gctgagatca ccaccacctt aaagggcaag gagtgtggca
     1141 ccacgatgac ctcctgcgtg tcctggccca cagcattcag atcctcttct gagatgagtt
     1201 tttgttcgtc gacctgcagt ttcagtgcct tgtcattcgg ttctttgaac agtgctttca
     1261 tttcgtctga cagttcgaaa ttcatgttca gggggttgtt aaagttgtac tccagcttat
     1321 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     1381 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     1441 agatggtgcg ctcctggatg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     1501 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg
     1561 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     1621 aggtggcatc gccctcaccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     1681 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgc
     1741 tccctccggt accgcccttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     1801 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttcgaa agtttggact
     1861 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     1921 gctggtagtg gtaggcgagc tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt
     1981 tcgccttgat gccgttcctc tgcttgtcgg ccatgatata gacgttgtgg ctcaccagaa
     2041 ttggattttt gaaccactta tcaaaccatt tttccgcttc accggaggtc tgcacctgag
     2101 cgatggtgtc atccatcagc tttttgaact gcggatcatc tttacgcaac atacaaccgt
     2161 aggcctcctg agactgcggc ttgccgacga tttcccagtt gtctggtttc ttcgctttcg
     2221 cacgttcacc ggccagcagc acgtcatcca tcataaaggc aacggcacga ccgctttcca
     2281 gggtgcggaa agagtcaccg tgatctttgg cgctgatgat gcgcatattc attttttgct
     2341 cttcattcag tttgttgagc aaaacttcag aggtagtgcc ggaagtgacg actacggctt
     2401 tgtctttcag gttggcaaaa tctttgatat cgccaccctt tttggtcaac aggcgcgtac
     2461 cgaccacgaa aatagtgtca gagaaagccg cctgtttttg gcgttcgacg ttgttggtgg
     2521 tagaaccaca ttcaaaatcg aaagtgccgt tttgcagcag tggaatacgg ttttgtgagg
     2581 taatcggaat cagttttacc tgcaagtccg gtttgttgag tttctttttc actgcttcaa
     2641 caatggcgtt ggagtaatcc tgcgagtaac ccaccacttt ttgctgattg tcgtaataag
     2701 agaaaggcac tgaagattca cggtgaccga cgacaatcac accgtttttg gcgattttgt
     2761 ccagcgtgct gcctgcggca gatctgtcac cagtggaacc tggaacccag agcagcagta
     2821 cccatagcag gagtgtgtct gtctccatgg tggcgatatc accggtggta ccactagtat
     2881 aacttcgtat aggatacttt atacgaagtt atcattggga ttcttcctat tttgatccaa
     2941 gcatcaccat cgaccctcta gtccagatct caccatcgac ccataacttc gtatagcata
     3001 cattatacga agttatgtcc ctcgaagagg ttcgaaacac tgaaaagggc gaattctgca
     3061 gatgatccac tcgagaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     3121 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3181 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3241 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3301 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3361 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3421 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3481 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     3541 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     3601 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     3661 tgggccgcct ccccgcatcg ataccgtcga cccgggcggc cgctcgagca tgcatctaga
     3721 gtcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     3781 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     3841 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     3901 tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat
     3961 gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta gcatggcggg
     4021 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     4081 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     4141 gcctcagtga gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag
     4201 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4261 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4321 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4381 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4441 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4501 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4561 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4621 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4681 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4741 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4801 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4861 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4921 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4981 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5041 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5101 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5161 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5221 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5281 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5341 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5401 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5461 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5521 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5581 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5641 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5701 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5761 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5821 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5881 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5941 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6001 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6061 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6121 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     6181 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6241 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     6301 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     6361 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     6421 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6481 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     6541 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6601 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6661 ctcttcgcta ttacgc
//
