LOCUS       sequence-201016-        6908 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DEFINITION  Tight affinity glutamate sensor ** A newer version of this sensor is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 6908)
  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 6908)
  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..6908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1171..1204
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1275..1308
                     /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      1333..1342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1339..1398
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     2367..2388
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2491..2512)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2977..3006
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3010..3156
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(3184..3217)
                     /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(3280..3313)
                     /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(3348..3367)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3429..3451)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3731..4008)
                     /label=chicken beta-actin promoter
     repeat_region   4321..4450
                     /label=AAV2 ITR
     rep_origin      4522..4977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4609..4628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4819..4840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5003..5107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5108..5176,5177..5968)
                     /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(5326..5345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6139..6727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6628..6647
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct tctcgagtgg atcatttgca gaattggccc
     1141 ttttcagtgt ttcgaacctc ttcgagggac ataacttcgt ataatgtatg ctatacgaag
     1201 ttatgggtcg atggtgagat ttggactaga gggtcgatgg tgatgcttgg atcaaaatag
     1261 gaagaatccc aatgataact tcgtataaag tatcctatac gaagttatac tagtggtacc
     1321 accggtgata tcgccaccat ggagacagac acactcctgc tatgggtact gctgctctgg
     1381 gttccaggtt ccactggtga cagatctgcc gcaggcagca cgctggacaa aatcgccaaa
     1441 aacggtgtga ttgtcgtcgg tcaccgtgaa tcttcagtgc ctttctctta ttacgacaat
     1501 cagcaaaaag tggtgggtta ctcgcaggat tactccaacg ccattgttga agcagtgaaa
     1561 aagaaactca acaaaccgga cttgcaggta aaactgattc cgattacctc acaaaaccgt
     1621 attccactgc tgcaaaacgg cactttcgat tttgaatgtg gttctaccac caacaacgtc
     1681 gaacgccaaa aacaggcggc tttctctgac actattttcg tggtcggtac gcgcctgttg
     1741 accaaaaagg gtggcgatat caaagatttt gccaacctga aagacaaagc cgtagtcgtc
     1801 acttccggca ctacctctga agttttgctc aacaaactga atgaagagca aaaaatgaat
     1861 atgcgcatca tcagcgccaa agatcacggt gactctttcc gcaccctgga aagcggtcgt
     1921 gccgttgcct ttatgatgga tgacagcctg ctggccggtg aacgtgcgaa agcgaagaaa
     1981 ccagacaact gggaaatcgt cggcaagccg cagtctcagg aggcctacgg ttgtatgttg
     2041 cgtaaagatg atccgcagtt caaaaagctg atggatgaca ccatcgctca ggtgcagacc
     2101 tccggtgaag cggaaaaatg gtttgataag tggttcaaaa atccaattct ggtgagccac
     2161 aacgtctata tcaccgccga caagcagaag aacggcatca aggcgaactt caagatccgc
     2221 cacaacgtgg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2281 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgtgctgagc
     2341 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2401 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat gagcaagggc
     2461 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2521 cacaagttca gcgtgcgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg
     2581 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2641 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2701 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcagcttcaa ggacgacggc
     2761 acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2821 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2881 tttaacaacc ccctgaacat gaatttcgaa ctgtcagacg aaatgaaagc actgttcaaa
     2941 gaaccgaatg acaaggcact gaaactgcag gtcgacgaac aaaaactcat ctcagaagag
     3001 gatctgaatg ctgtgggcca ggacacgcag gaggtcatcg tggtgccaca ctccttgccc
     3061 tttaaggtgg tggtgatctc agccatcctg gccctggtgg tgctcaccat catctccctt
     3121 atcatcctca tcatgctttg gcagaagaag ccacgttagg atatcggatc cgctagcact
     3181 agtataactt cgtatagcat acattatacg aagttatggg tcgatggtga tgcttggcaa
     3241 ttcgggtcga tggtgaagca ttggtcttcc tattttgata taacttcgta taggatactt
     3301 tatacgaagt tatggggatc caattctttg ccaaaatgat gagacagcac aataaccagc
     3361 acgttgccca ggagctgtag gaaaaagaag aaggcatgaa catggttagc agaggctcta
     3421 gagccgccgg tcacacgcca gaagccgaac cccgccctgc cccgtccccc ccgaaggcag
     3481 ccgtcccccc gcggacagcc ccgagccgct ccccccgcac aaagggccct cccggagccc
     3541 ctcaaggctt tcacgcagcc acagaaaaga aacaagccgt cattaaacca agcgctaatt
     3601 acagcccgga ggagaagggc cgtcccgccc gctcacctgt gggagtaacg cggtcagtca
     3661 gagccggggc gggcggcgcg aggcggcggc ggagcggggc acggggcgaa ggcagcgtcg
     3721 cagcgactcc cgcccgccgc gcgcttcgct ttttataggg ccgccgccgc cgccgcctcg
     3781 ccataaaagg aaactttcgg agcgcgccgc tctgattggc tgccgccgca cctctccgcc
     3841 tcgccccgcc ccgcccctcg ccccgccccg ccccgcctgg cgcgcgcccc cccccccccc
     3901 ccgcccccat cgctgcacaa aataattaaa aaataaataa atacaaaatt gggggtgggg
     3961 agggggggga gatggggaga gtgaagcaga acgtggggct cacctcgacc atggtaatag
     4021 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     4081 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     4141 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     4201 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatggta
     4261 cgatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
     4321 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     4381 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4441 gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     4501 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4561 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4621 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4681 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4741 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4801 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4861 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4921 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     4981 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5041 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5101 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5161 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5221 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5281 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5341 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5401 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5461 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5521 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5581 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5641 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5701 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5761 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5821 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5881 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5941 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6001 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6061 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6121 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6181 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6241 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6301 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6361 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6421 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6481 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6541 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6601 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6661 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6721 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6781 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6841 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6901 agcggaag
//
