LOCUS       sequence-201031-        7643 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DEFINITION  Tight affinity glutamate sensor ** A newer version of this sensor is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 7643)
  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 7643)
  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.mRuby3
FEATURES             Location/Qualifiers
     source          1..7643
                     /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(3919..3952)
                     /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(4015..4048)
                     /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(4083..4102)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4164..4186)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(4466..4743)
                     /label=chicken beta-actin promoter
     repeat_region   5056..5185
                     /label=AAV2 ITR
     rep_origin      5257..5712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5344..5363)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5554..5575
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5738..5842
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5843..5911,5912..6703)
                     /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(6061..6080)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6874..7462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7363..7382
                     /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 ccacgttcta gaggctccgg aaatgctgtg
     3181 ggccaggaca cgcaggagcg tatggtgtct aagggcgaag agctgatcaa ggaaaatatg
     3241 cgtatgaagg tggtcatgga aggttcggtc aacggccacc aattcaaatg cacaggtgaa
     3301 ggagaaggca ggccgtacga gggagtgcaa accatgagga tcaaagtcat cgagggagga
     3361 cccctgccat ttgcctttga cattcttgcc acgtcgttca tgtatggcag ccgtactttt
     3421 atcaagtacc cggcagacat tcctgatttc tttaaacagt cctttcctga gggttttact
     3481 tgggaaagag ttacgagata cgaagatggt ggagtcgtca ccgtcacgca ggacaccagc
     3541 cttgaggatg gcgaactcgt ttacaacgtc aaagtcagag gggtaaactt tccctccaat
     3601 ggtcccgtga tgcagaagaa gaccaagggt tgggagccta atacagagat gatgtatcca
     3661 gcagatggtg gtctgagggg atacactgat attgcactga aagttgatgg tggtggccat
     3721 ctgcattgca atttcgtaac aacttacagg tcaaaaaaga ccgtcgggaa catcaagatg
     3781 cccggtgtcc atgccgttga tcaccgcctg gaaaggatag aggaaagtga caatgaaacg
     3841 tacgtagtac aacgcgaagt cgcagttgcc aagtactcca atcttggtgg ttaggatatc
     3901 ggatccgcta gcactagtat aacttcgtat agcatacatt atacgaagtt atgggtcgat
     3961 ggtgatgctt ggcaattcgg gtcgatggtg aagcattggt cttcctattt tgatataact
     4021 tcgtatagga tactttatac gaagttatgg ggatccaatt ctttgccaaa atgatgagac
     4081 agcacaataa ccagcacgtt gcccaggagc tgtaggaaaa agaagaaggc atgaacatgg
     4141 ttagcagagg ctctagagcc gccggtcaca cgccagaagc cgaaccccgc cctgccccgt
     4201 cccccccgaa ggcagccgtc cccccgcgga cagccccgag ccgctccccc cgcacaaagg
     4261 gccctcccgg agcccctcaa ggctttcacg cagccacaga aaagaaacaa gccgtcatta
     4321 aaccaagcgc taattacagc ccggaggaga agggccgtcc cgcccgctca cctgtgggag
     4381 taacgcggtc agtcagagcc ggggcgggcg gcgcgaggcg gcggcggagc ggggcacggg
     4441 gcgaaggcag cgtcgcagcg actcccgccc gccgcgcgct tcgcttttta tagggccgcc
     4501 gccgccgccg cctcgccata aaaggaaact ttcggagcgc gccgctctga ttggctgccg
     4561 ccgcacctct ccgcctcgcc ccgccccgcc cctcgccccg ccccgccccg cctggcgcgc
     4621 gccccccccc cccccccgcc cccatcgctg cacaaaataa ttaaaaaata aataaataca
     4681 aaattggggg tggggagggg ggggagatgg ggagagtgaa gcagaacgtg gggctcacct
     4741 cgaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     4801 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     4861 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     4921 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     4981 ttgacgtcaa tggtacgatc ttcctagagc atggctacgt agataagtag catggcgggt
     5041 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     5101 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     5161 cctcagtgag cgagcgagcg cgcagctggc gtaatagcga agaggcccgc accgatcgcc
     5221 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     5281 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5341 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5401 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5461 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5521 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5581 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     5641 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5701 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5761 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5821 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5881 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5941 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6001 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6061 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6121 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6181 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6241 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6301 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6361 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6421 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6481 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6541 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6601 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6661 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6721 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6781 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6841 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6901 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6961 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7021 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7081 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7141 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7201 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7261 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7321 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7381 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7441 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7501 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7561 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     7621 gtcagtgagc gaggaagcgg aag
//
