LOCUS       sequence-204258-        7483 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Weak affinity glutamate sensor ** A newer version of this sensor is 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      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7483)
  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 7483)
  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-iGluSnFR.S72A.mRuby3
FEATURES             Location/Qualifiers
     source          1..7483
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     repeat_region   459..588
                     /label=AAV2 ITR
     rep_origin      660..1115
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(747..766)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     957..978
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1141..1245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1246..1314,1315..2106)
                     /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(1464..1483)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2277..2865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2766..2785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3393..3514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3430..3449)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3484..3503
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3557..4145
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3663..3674
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4072..4092
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    4217..4250
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4321..4354
                     /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      4379..4388
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     4385..4444
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     5413..5434
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5537..5558)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             6023..6052
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6056..6202
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(6965..6998)
                     /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(7061..7094)
                     /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(7129..7148)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(7210..7232)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 ccccgccccg ccccgcctgg cgcgcgcccc cccccccccc ccgcccccat cgctgcacaa
       61 aataattaaa aaataaataa atacaaaatt gggggtgggg agggggggga gatggggaga
      121 gtgaagcaga acgtggggct cacctcgacc atggtaatag cgatgactaa tacgtagatg
      181 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
      241 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
      301 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
      361 gttactatgg gaacatacgt cattattgac gtcaatggta ctcttcctag agcatggcta
      421 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
      481 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
      541 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct ggcgtaatag
      601 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
      661 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
      721 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
      781 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
      841 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
      901 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
      961 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1021 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1081 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     1141 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1201 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1261 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1321 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1381 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1441 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     1501 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     1561 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1621 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1681 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1741 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1801 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1861 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1921 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1981 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2041 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2101 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     2161 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2221 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2281 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2341 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2401 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2461 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2521 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2581 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2641 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2701 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2761 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2821 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2881 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2941 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3001 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3061 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctgcgcgct cgctcgctca
     3121 ctgaggccgc ccgggmwams cccygmsgcm gccsrggcaa agcccgggcg gcctcagtga
     3181 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa
     3241 tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagatc cttatcgatt
     3301 ttaccacatt tgtagaggtt ttacttgctt taaaaaacct cccacatctc cccctgaacc
     3361 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3421 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3481 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgctcgaag cggccgcccg
     3541 ggtcgacggt atcgatgcgg ggaggcggcc caaagggaga tccgactcgt ctgagggcga
     3601 aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct
     3661 ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca
     3721 caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga attgtcagtg
     3781 cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg
     3841 gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc
     3901 caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg cctgacaacg
     3961 ggccacaact cctcataaag agacagcaac caggatttat acaaggagga gaaaatgaaa
     4021 gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta
     4081 aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta atccagaggt
     4141 tgattatcga taagcttctc gagtggatca tttgcagaat tggccctttt cagtgtttcg
     4201 aacctcttcg agggacataa cttcgtataa tgtatgctat acgaagttat gggtcgatgg
     4261 tgagatttgg actagagggt cgatggtgat gcttggatca aaataggaag aatcccaatg
     4321 ataacttcgt ataaagtatc ctatacgaag ttatactagt ggtaccaccg gtgatatcgc
     4381 caccatggag acagacacac tcctgctatg ggtactgctg ctctgggttc caggttccac
     4441 tggtgacaga tctgccgcag gcagcacgct ggacaaaatc gccaaaaacg gtgtgattgt
     4501 cgtcggtcac cgtgaatctt cagtgccttt ctcttattac gacaatcagc aaaaagtggt
     4561 gggttactcg caggattact ccaacgccat tgttgaagca gtgaaaaaga aactcaacaa
     4621 accggacttg caggtaaaac tgattccgat taccgcacaa aaccgtattc cactgctgca
     4681 aaacggcact ttcgattttg aatgtggttc taccaccaac aacgtcgaac gccaaaaaca
     4741 ggcggctttc tctgacacta ttttcgtggt cggtacgcgc ctgttgacca aaaagggtgg
     4801 cgatatcaaa gattttgcca acctgaaaga caaagccgta gtcgtcactt ccggcactac
     4861 ctctgaagtt ttgctcaaca aactgaatga agagcaaaaa atgaatatgc gcatcatcag
     4921 cgccaaagat cacggtgact ctttccgcac cctggaaagc ggtcgtgccg ttgcctttat
     4981 gatggatgac gtgctgctgg ccggtgaacg tgcgaaagcg aagaaaccag acaactggga
     5041 aatcgtcggc aagccgcagt ctcaggaggc ctacggttgt atgttgcgta aagatgatcc
     5101 gcagttcaaa aagctgatgg atgacaccat cgctcaggtg cagacctccg gtgaagcgga
     5161 aaaatggttt gataagtggt tcaaaaatcc aattctggtg agccacaacg tctatatcac
     5221 cgccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca acgtggagga
     5281 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     5341 gctgctgccc gacaaccact acctgagcac ccagtccgtg ctgagcaaag accccaacga
     5401 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     5461 ggacgagctg tacaagggcg gtaccggagg gagcatgagc aagggcgagg agctgttcac
     5521 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     5581 gcgcggcgag ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac
     5641 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     5701 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     5761 cgaaggctac gtccaggagc gcaccatcag cttcaaggac gacggcacct acaagacccg
     5821 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     5881 cttcaaggag gacggcaaca tcctggggca caagctggag tacaacttta acaaccccct
     5941 gaacatgaat ttcgaactgt cagacgaaat gaaagcactg ttcaaagaac cgaatgacaa
     6001 ggcactgaaa ctgcaggtcg acgaacaaaa actcatctca gaagaggatc tgaatgctgt
     6061 gggccaggac acgcaggagg tcatcgtggt gccacactcc ttgcccttta aggtggtggt
     6121 gatctcagcc atcctggccc tggtggtgct caccatcatc tcccttatca tcctcatcat
     6181 gctttggcag aagaagccac gttctagagg ctccggaaat gctgtgggcc aggacacgca
     6241 ggagcgtatg gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt
     6301 catggaaggt tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag aaggcaggcc
     6361 gtacgaggga gtgcaaacca tgaggatcaa agtcatcgag ggaggacccc tgccatttgc
     6421 ctttgacatt cttgccacgt cgttcatgta tggcagccgt acttttatca agtacccggc
     6481 agacattcct gatttcttta aacagtcctt tcctgagggt tttacttggg aaagagttac
     6541 gagatacgaa gatggtggag tcgtcaccgt cacgcaggac accagccttg aggatggcga
     6601 actcgtttac aacgtcaaag tcagaggggt aaactttccc tccaatggtc ccgtgatgca
     6661 gaagaagacc aagggttggg agcctaatac agagatgatg tatccagcag atggtggtct
     6721 gaggggatac actgatattg cactgaaagt tgatggtggt ggccatctgc attgcaattt
     6781 cgtaacaact tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg gtgtccatgc
     6841 cgttgatcac cgcctggaaa ggatagagga aagtgacaat gaaacgtacg tagtacaacg
     6901 cgaagtcgca gttgccaagt actccaatct tggtggttag gatatcggat ccgctagcac
     6961 tagtataact tcgtatagca tacattatac gaagttatgg gtcgatggtg atgcttggca
     7021 attcgggtcg atggtgaagc attggtcttc ctattttgat ataacttcgt ataggatact
     7081 ttatacgaag ttatggggat ccaattcttt gccaaaatga tgagacagca caataaccag
     7141 cacgttgccc aggagctgta ggaaaaagaa gaaggcatga acatggttag cagaggctct
     7201 agagccgccg gtcacacgcc agaagccgaa ccccgccctg ccccgtcccc cccgaaggca
     7261 gccgtccccc cgcggacagc cccgagccgc tccccccgca caaagggccc tcccggagcc
     7321 cctcaaggct ttcacgcagc cacagaaaag aaacaagccg tcattaaacc aagcgctaat
     7381 tacagcccgg aggagaaggg ccgtcccgcc cgctcacctg tgggagtaac gcggtcagtc
     7441 agagccgggg cgggcggcgc gaggcggcgg cggagcgggg cac
//
