LOCUS       sequence-181244-        6878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre dependent AAV expression of glutamate sensor from GFAP 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            ** A newer version of this sensor is available.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 6878)
  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 6878)
  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.CAG.Flex.iGluSnFr.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        441..718
                     /label=chicken beta-actin promoter
     primer_bind     997..1019
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1081..1100
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1135..1168
                     /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    1231..1264
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1292..1438)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1442..1471)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1677..1696
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1936..1957
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2063..2084)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(3053..3112)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      3109..3118
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3143..3176)
                     /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(3247..3280)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3322..3910
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3375..3395)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3793..3804)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3953..4074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3964..3983)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4018..4037
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4236..4365
                     /label=AAV2 ITR
     rep_origin      complement(4602..5190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4682..4701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5361..6152,6153..6221))
                     /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     5984..6003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6222..6326)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6352..6807)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6489..6510)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6701..6720
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      601 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      661 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      721 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      781 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      841 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      901 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg
      961 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1021 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1081 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattggat ccccataact
     1141 tcgtataaag tatcctatac gaagttatat caaaatagga agaccaatgc ttcaccatcg
     1201 acccgaattg ccaagcatca ccatcgaccc ataacttcgt ataatgtatg ctatacgaag
     1261 ttatactagt gctagcggat ccgatatcct aacgtggctt cttctgccaa agcatgatga
     1321 ggatgataag ggagatgatg gtgagcacca ccagggccag gatggctgag atcaccacca
     1381 ccttaaaggg caaggagtgt ggcaccacga tgacctcctg cgtgtcctgg cccacagcat
     1441 tcagatcctc ttctgagatg agtttttgtt cgtcgacctg cagtttcagt gccttgtcat
     1501 tcggttcttt gaacagtgct ttcatttcgt ctgacagttc gaaattcatg ttcagggggt
     1561 tgttaaagtt gtactccagc ttatgcccca ggatgttgcc gtcctccttg aagtcgatgc
     1621 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     1681 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gatgtagcct tcgggcatgg
     1741 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
     1801 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     1861 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc accctcgccg gacacgctga
     1921 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     1981 gctcctcgcc cttgctcacc atgctccctc cggtaccgcc cttgtacagc tcgtccatgc
     2041 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     2101 tggggtcttt cgaaagtttg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
     2161 ggccgtcgcc gatgggggtg ttctgctggt agtggtaggc gagctgcacg ccgccgtcct
     2221 cgatgttgtg gcggatcttg aagttcgcct tgatgccgtt cctctgcttg tcggccatga
     2281 tatagacgtt gtggctcacc agaattggat ttttgaacca cttatcaaac catttttccg
     2341 cttcaccgga ggtctgcacc tgagcgatgg tgtcatccat cagctttttg aactgcggat
     2401 catctttacg caacatacaa ccgtaggcct cctgagactg cggcttgccg acgatttccc
     2461 agttgtctgg tttcttcgct ttcgcacgtt caccggccag cagcacgtca tccatcataa
     2521 aggcaacggc acgaccgctt tccagggtgc ggaaagagtc accgtgatct ttggcgctga
     2581 tgatgcgcat attcattttt tgctcttcat tcagtttgtt gagcaaaact tcagaggtag
     2641 tgccggaagt gacgactacg gctttgtctt tcaggttggc aaaatctttg atatcgccac
     2701 cctttttggt caacaggcgc gtaccgacca cgaaaatagt gtcagagaaa gccgcctgtt
     2761 tttggcgttc gacgttgttg gtggtagaac cacattcaaa atcgaaagtg ccgttttgca
     2821 gcagtggaat acggttttgt gaggtaatcg gaatcagttt tacctgcaag tccggtttgt
     2881 tgagtttctt tttcactgct tcaacaatgg cgttggagta atcctgcgag taacccacca
     2941 ctttttgctg attgtcgtaa taagagaaag gcactgaaga ttcacggtga ccgacgacaa
     3001 tcacaccgtt tttggcgatt ttgtccagcg tgctgcctgc ggcagatctg tcaccagtgg
     3061 aacctggaac ccagagcagc agtacccata gcaggagtgt gtctgtctcc atggtggcga
     3121 tatcaccggt ggtaccacta gtataacttc gtataggata ctttatacga agttatcatt
     3181 gggattcttc ctattttgat ccaagcatca ccatcgaccc tctagtccag atctcaccat
     3241 cgacccataa cttcgtatag catacattat acgaagttat gtccctcgaa gaggttcgaa
     3301 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     3361 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3421 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3481 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3541 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3601 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3661 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3721 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3781 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3841 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3901 gcctccccgc atcgataccg tcgacccggg cggccgcttc gagcagacat gataagatac
     3961 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     4021 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4081 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     4141 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt
     4201 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     4261 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4321 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc
     4381 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4441 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4501 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4561 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4621 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4681 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4741 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4801 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4861 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4921 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4981 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5041 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5101 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5161 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5221 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5281 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5341 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5401 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5461 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5521 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5581 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5641 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5701 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5761 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5821 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5881 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5941 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6001 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6061 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6121 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6181 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6241 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6301 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     6361 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     6421 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     6481 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     6541 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     6601 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     6661 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     6721 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     6781 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     6841 aagggcgatc ggtgcgggcc tcttcgctat tacgccag
//
