LOCUS       sequence-189881-        6659 bp    DNA     circular SYN 14-APR-2025
DEFINITION  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 ** A newer 
            version of this sensor is available.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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6659)
  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 6659)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pENN.AAV.GFAP.iGluSnFr.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    85..106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(121..126,127..144,145..151)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    159..175
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     164..186
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     183..199
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     183..199
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1140..1202
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     primer_bind     2168..2189
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2295..2316)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2556..2575)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2781..2810
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2814..2960
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2976..3564
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3029..3049)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3447..3458)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3567..3583)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3568..3584)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3613..3734
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3624..3643)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3678..3697
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3945..4074
                     /label=AAV2 ITR
     primer_bind     complement(4093..4110)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4093..4109)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4102..4124)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4251..4706
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4338..4357)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4548..4569
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4732..4836
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4837..4905,4906..5697)
                     /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"
     rep_origin      5868..6456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6357..6376
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
       61 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      121 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      181 cacaggaaac agctatgacc atgattacgc cagatttaat taaggcctta attaggctgc
      241 gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcggcctcag tgagcgagcg
      301 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa
      361 cccgccatgc tacttatcta cgtagccatg ctctaggaag atcggaattc gcccttaagc
      421 tagccgagct cggtaccaga tctaacatat cctggtgtgg agtaggggac gctgctctga
      481 cagaggctcg ggggcctgag ctggctctgt gagctgggga ggaggcagac agccaggcct
      541 tgtctgcaag cagacctggc agcattgggc tggccgcccc ccagggcctc ctcttcatgc
      601 ccagtgaatg actcaccttg gcacagacac aatgttcggg gtgggcacag tgcctgcttc
      661 ccgccgcacc ccagcccccc tcaaatgcct tccgagaagc ccattgagca gggggcttgc
      721 attgcacccc agcctgacag cctggcatct tgggataaaa gcagcacagc cccctagggg
      781 ctgcccttgc tgtgtggcgc caccggcggt ggagaacaag gctctattca gcctgtgccc
      841 aggaaagggg atcaggggat gcccaggcat ggacagtggg tggcaggggg ggagaggagg
      901 gctgtctgct tcccagaagt ccaaggacac aaatgggtga ggggagagct ctccccatag
      961 ctgggctgcg gcccaacccc accccctcag gctatgccag ggggtgttgc caggggcacc
     1021 cgggcatcgc cagtctagcc cactccttca taaagccctc gcatcccagg agcgagcaga
     1081 gccagagcag gttggagagg agacgcatca cctccgctgc tcgcggggat cccgccacca
     1141 tggagacaga cacactcctg ctatgggtac tgctgctctg ggttccaggt tccactggtg
     1201 acagatctgc cgcaggcagc acgctggaca aaatcgccaa aaacggtgtg attgtcgtcg
     1261 gtcaccgtga atcttcagtg cctttctctt attacgacaa tcagcaaaaa gtggtgggtt
     1321 actcgcagga ttactccaac gccattgttg aagcagtgaa aaagaaactc aacaaaccgg
     1381 acttgcaggt aaaactgatt ccgattacct cacaaaaccg tattccactg ctgcaaaacg
     1441 gcactttcga ttttgaatgt ggttctacca ccaacaacgt cgaacgccaa aaacaggcgg
     1501 ctttctctga cactattttc gtggtcggta cgcgcctgtt gaccaaaaag ggtggcgata
     1561 tcaaagattt tgccaacctg aaagacaaag ccgtagtcgt cacttccggc actacctctg
     1621 aagttttgct caacaaactg aatgaagagc aaaaaatgaa tatgcgcatc atcagcgcca
     1681 aagatcacgg tgactctttc cgcaccctgg aaagcggtcg tgccgttgcc tttatgatgg
     1741 atgacgtgct gctggccggt gaacgtgcga aagcgaagaa accagacaac tgggaaatcg
     1801 tcggcaagcc gcagtctcag gaggcctacg gttgtatgtt gcgtaaagat gatccgcagt
     1861 tcaaaaagct gatggatgac accatcgctc aggtgcagac ctccggtgaa gcggaaaaat
     1921 ggtttgataa gtggttcaaa aatccaattc tggtgagcca caacgtctat atcatggccg
     1981 acaagcagag gaacggcatc aaggcgaact tcaagatccg ccacaacatc gaggacggcg
     2041 gcgtgcagct cgcctaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2101 tgcccgacaa ccactacctg agcacccagt ccaaactttc gaaagacccc aacgagaagc
     2161 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2221 agctgtacaa gggcggtacc ggagggagca tggtgagcaa gggcgaggag ctgttcaccg
     2281 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2341 ccggcgaggg tgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2401 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     2461 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2521 aaggctacat ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2581 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2641 tcaaggagga cggcaacatc ctggggcata agctggagta caactttaac aaccccctga
     2701 acatgaattt cgaactgtca gacgaaatga aagcactgtt caaagaaccg aatgacaagg
     2761 cactgaaact gcaggtcgac gaacaaaaac tcatctcaga agaggatctg aatgctgtgg
     2821 gccaggacac gcaggaggtc atcgtggtgc cacactcctt gccctttaag gtggtggtga
     2881 tctcagccat cctggccctg gtggtgctca ccatcatctc ccttatcatc ctcatcatgc
     2941 tttggcagaa gaagccacgt tagaagctta tcgataatca acctctggat tacaaaattt
     3001 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3061 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3121 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3181 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3241 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3301 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3361 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3421 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3481 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3541 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgacccggg cggccgcttc
     3601 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     3661 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     3721 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     3781 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc
     3841 ttcctagagc atggctactc gagttaaggg cgaattcccg ataaggatct tcctagagca
     3901 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
     3961 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4021 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagccttaa
     4081 ttaacctaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     4141 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     4201 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     4261 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4321 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4381 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4441 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4501 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4561 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4621 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4681 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     4741 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4801 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4861 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4921 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4981 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5041 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5101 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5161 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5221 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5281 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5341 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5401 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5461 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5521 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5581 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5641 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5701 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5761 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5821 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5881 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5941 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6001 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6061 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6121 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6181 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6241 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6301 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6361 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6421 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6481 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6541 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6601 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcc
//
