LOCUS       sequence-189980-        6834 bp    DNA     linear   UNA 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            ** A newer version of this sensor is available.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      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6834)
  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 6834)
  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..6834
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     375..397
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     459..478
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    513..546
                     /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    609..642
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(670..816)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(820..849)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1055..1074
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1314..1335
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1441..1462)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2431..2490)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2487..2496
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2521..2554)
                     /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(2625..2658)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2700..3288
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2753..2773)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3171..3182)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3331..3452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3342..3361)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3396..3415
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3614..3743
                     /label=AAV2 ITR
     rep_origin      complement(3980..4568)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4060..4079)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4739..5530,5531..5599))
                     /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     5362..5381
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5600..5704)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5730..6185)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5867..5888)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6079..6098
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6257..6386
                     /label=AAV2 ITR
ORIGIN
        1 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
       61 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      121 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      181 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      241 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
      301 cctttgtgcg gggggagcgg ctcggggctg tccgcggggg gacggctgcc ttcggggggg
      361 acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa
      421 ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct
      481 gtctcatcat tttggcaaag aattggatcc ccataacttc gtataaagta tcctatacga
      541 agttatatca aaataggaag accaatgctt caccatcgac ccgaattgcc aagcatcacc
      601 atcgacccat aacttcgtat aatgtatgct atacgaagtt atactagtgc tagcggatcc
      661 gatatcctaa cgtggcttct tctgccaaag catgatgagg atgataaggg agatgatggt
      721 gagcaccacc agggccagga tggctgagat caccaccacc ttaaagggca aggagtgtgg
      781 caccacgatg acctcctgcg tgtcctggcc cacagcattc agatcctctt ctgagatgag
      841 tttttgttcg tcgacctgca gtttcagtgc cttgtcattc ggttctttga acagtgcttt
      901 catttcgtct gacagttcga aattcatgtt cagggggttg ttaaagttgt actccagctt
      961 atgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     1021 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     1081 gaagatggtg cgctcctgga tgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     1141 cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag
     1201 ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc
     1261 gtaggtggca tcgccctcac cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     1321 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     1381 gctccctccg gtaccgccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     1441 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttcg aaagtttgga
     1501 ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     1561 ctgctggtag tggtaggcga gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa
     1621 gttcgccttg atgccgttcc tctgcttgtc ggccatgata tagacgttgt ggctcaccag
     1681 aattggattt ttgaaccact tatcaaacca tttttccgct tcaccggagg tctgcacctg
     1741 agcgatggtg tcatccatca gctttttgaa ctgcggatca tctttacgca acatacaacc
     1801 gtaggcctcc tgagactgcg gcttgccgac gatttcccag ttgtctggtt tcttcgcttt
     1861 cgcacgttca ccggccagca gcacgtcatc catcataaag gcaacggcac gaccgctttc
     1921 cagggtgcgg aaagagtcac cgtgatcttt ggcgctgatg atgcgcatat tcattttttg
     1981 ctcttcattc agtttgttga gcaaaacttc agaggtagtg ccggaagtga cgactacggc
     2041 tttgtctttc aggttggcaa aatctttgat atcgccaccc tttttggtca acaggcgcgt
     2101 accgaccacg aaaatagtgt cagagaaagc cgcctgtttt tggcgttcga cgttgttggt
     2161 ggtagaacca cattcaaaat cgaaagtgcc gttttgcagc agtggaatac ggttttgtga
     2221 ggtaatcgga atcagtttta cctgcaagtc cggtttgttg agtttctttt tcactgcttc
     2281 aacaatggcg ttggagtaat cctgcgagta acccaccact ttttgctgat tgtcgtaata
     2341 agagaaaggc actgaagatt cacggtgacc gacgacaatc acaccgtttt tggcgatttt
     2401 gtccagcgtg ctgcctgcgg cagatctgtc accagtggaa cctggaaccc agagcagcag
     2461 tacccatagc aggagtgtgt ctgtctccat ggtggcgata tcaccggtgg taccactagt
     2521 ataacttcgt ataggatact ttatacgaag ttatcattgg gattcttcct attttgatcc
     2581 aagcatcacc atcgaccctc tagtccagat ctcaccatcg acccataact tcgtatagca
     2641 tacattatac gaagttatgt ccctcgaaga ggttcgaatt cgatatcaag cttatcgata
     2701 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2761 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2821 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2881 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2941 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3001 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3061 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3121 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3181 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3241 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     3301 gacccgggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     3361 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3421 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3481 ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3541 gtaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta
     3601 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3661 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3721 tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3781 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3841 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3901 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3961 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4021 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4081 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4141 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4201 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4261 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4321 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4381 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4441 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4501 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4561 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4621 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4681 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4741 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4801 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4861 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4921 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4981 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5041 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5101 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5161 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5221 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5281 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5341 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5401 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5461 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5521 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5581 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5641 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5701 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     5761 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     5821 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     5881 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     5941 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6001 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     6061 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6121 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     6181 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6241 ttcgctatta cgccagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
     6301 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     6361 ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc
     6421 tacgtagcca tgctctagga agagtaccat tgacgtcaat aatgacgtat gttcccatag
     6481 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     6541 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     6601 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     6661 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
     6721 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
     6781 tattttgtgc agcgatgggg gcgggggggg ggggggggcg cgcgccaggc gggg
//
