LOCUS       sequence-214933-        6905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tight affinity glutamate sensor ** A newer version of this sensor is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      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-iGluSnFR.A184S
FEATURES             Location/Qualifiers
     source          1..6905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   38..167
                     /label=AAV2 ITR
     promoter        478..755
                     /label=chicken beta-actin promoter
     primer_bind     1034..1056
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1118..1137
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1172..1205
                     /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    1268..1301
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1329..1475)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1479..1508)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1973..1994
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2097..2118)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(3087..3146)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      3143..3152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3177..3210)
                     /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(3281..3314)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3386..3974
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3439..3459)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3857..3868)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4017..4138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4028..4047)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4082..4101
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4300..4429
                     /label=AAV2 ITR
     rep_origin      complement(4666..5254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4746..4765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5425..6216,6217..6285))
                     /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     6048..6067
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6286..6390)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6416..6871)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6553..6574)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6765..6784
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agggcgatcg gtgcgggcct cttcgctatt acgccagctg cgcgctcgct cgctcactga
       61 ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
      121 gcgagcgcgc agagagggag tggccaactc catcactagg ggttccttgt agttaatgat
      181 taacccgcca tgctacttat ctacgtagcc atgctctagg aagagtacca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
      481 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
      541 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc
      601 gcgcgccrgg sggggcgggg sggggcgrgg ggsggggcgg ggcgaggcgg agaggtgcgg
      661 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
      721 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
      781 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
      841 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
      901 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
      961 ctttgtgcgg ggggagcggc tcggggctgt ccgcgggggg acggctgcct tcggggggga
     1021 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac
     1081 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg
     1141 tctcatcatt ttggcaaaga attggatccc cataacttcg tataaagtat cctatacgaa
     1201 gttatatcaa aataggaaga ccaatgcttc accatcgacc cgaattgcca agcatcacca
     1261 tcgacccata acttcgtata atgtatgcta tacgaagtta tactagtgct agcggatccg
     1321 atatcctaac gtggcttctt ctgccaaagc atgatgagga tgataaggga gatgatggtg
     1381 agcaccacca gggccaggat ggctgagatc accaccacct taaagggcaa ggagtgtggc
     1441 accacgatga cctcctgcgt gtcctggccc acagcattca gatcctcttc tgagatgagt
     1501 ttttgttcgt cgacctgcag tttcagtgcc ttgtcattcg gttctttgaa cagtgctttc
     1561 atttcgtctg acagttcgaa attcatgttc agggggttgt taaagttgta ctccagcttg
     1621 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     1681 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt aggtgccgtc gtccttgaag
     1741 ctgatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     1801 ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg
     1861 gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg
     1921 ttggtggcat cgccctcgcc ctcgccgcgc acgctgaact tgtggccgtt tacgtcgccg
     1981 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcatgctc
     2041 cctccggtac cgcccttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg
     2101 aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag cacggactgg
     2161 gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc
     2221 tggtagtggt cggcgagctg cacgctgccg tcctccacgt tgtggcggat cttgaagttc
     2281 gccttgatgc cgttcttctg cttgtcggcg gtgatataga cgttgtggct caccagaatt
     2341 ggatttttga accacttatc aaaccatttt tccgcttcac cggaggtctg cacctgagcg
     2401 atggtgtcat ccatcagctt tttgaactgc ggatcatctt tacgcaacat acaaccgtag
     2461 gcctcctgag actgcggctt gccgacgatt tcccagttgt ctggtttctt cgctttcgca
     2521 cgttcaccgg ccagcaggct gtcatccatc ataaaggcaa cggcacgacc gctttccagg
     2581 gtgcggaaag agtcaccgtg atctttggcg ctgatgatgc gcatattcat tttttgctct
     2641 tcattcagtt tgttgagcaa aacttcagag gtagtgccgg aagtgacgac tacggctttg
     2701 tctttcaggt tggcaaaatc tttgatatcg ccaccctttt tggtcaacag gcgcgtaccg
     2761 accacgaaaa tagtgtcaga gaaagccgcc tgtttttggc gttcgacgtt gttggtggta
     2821 gaaccacatt caaaatcgaa agtgccgttt tgcagcagtg gaatacggtt ttgtgaggta
     2881 atcggaatca gttttacctg caagtccggt ttgttgagtt tctttttcac tgcttcaaca
     2941 atggcgttgg agtaatcctg cgagtaaccc accacttttt gctgattgtc gtaataagag
     3001 aaaggcactg aagattcacg gtgaccgacg acaatcacac cgtttttggc gattttgtcc
     3061 agcgtgctgc ctgcggcaga tctgtcacca gtggaacctg gaacccagag cagcagtacc
     3121 catagcagga gtgtgtctgt ctccatggtg gcgatatcac cggtggtacc actagtataa
     3181 cttcgtatag gatactttat acgaagttat cattgggatt cttcctattt tgatccaagc
     3241 atcaccatcg accctctagt ccaaatctca ccatcgaccc ataacttcgt atagcataca
     3301 ttatacgaag ttatgtccct cgaagaggtt cgaaacactg aaaagggcca attctgcaaa
     3361 tgatccactc gagaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     3421 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3481 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3541 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3601 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3661 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3721 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3781 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3841 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3901 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3961 ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg cttcgagcag acatgataag
     4021 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4081 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4141 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     4201 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct
     4261 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     4321 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4381 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg
     4441 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4501 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4561 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4621 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4681 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4741 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4801 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4861 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4921 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4981 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5041 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5101 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5161 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5221 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5281 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5341 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5401 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5461 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5521 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5581 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5641 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5701 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5761 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5821 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5881 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5941 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6001 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6061 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6121 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6181 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6241 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6301 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6361 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     6421 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     6481 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     6541 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     6601 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     6661 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     6721 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     6781 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6841 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     6901 tggga
//
