LOCUS       sequence-204270-        7025 bp    DNA     circular SYN 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DEFINITION  Weak affinity glutamate sensor ** A newer version of this sensor is 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7025)
  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 7025)
  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.hSynap-FLEX.SF-iGluSnFR.S72A
FEATURES             Location/Qualifiers
     source          1..7025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   53..182
                     /label=AAV2 ITR
     intron          775..907
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     890..909
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    920..953
                     /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    1016..1049
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1077..1223)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1227..1256)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1721..1742
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1845..1866)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2835..2894)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2891..2900
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2925..2958)
                     /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(3029..3062)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3134..3722
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3187..3207)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3605..3616)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3800..3921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3811..3830)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3865..3884
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(4363..4381)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4364..4381)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4399..4415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4399..4415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4412..4434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4423..4439
                     /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)."
     promoter        complement(join(4447..4453,4454..4471,4472..4477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4492..4513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4801..5389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4881..4900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5560..6351,6352..6420))
                     /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     6183..6202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6421..6525)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6551..7006)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6688..6709)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6900..6919
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctgcgcgc
       61 tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc
      121 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
      181 cttgtagtta atgattaacc cgccatgcta cttatctacg tagccatgct ctaggaagat
      241 ccgagctcca gtgtgctgga attcgccctt ttcagtattt aaattgacga ccgaccccga
      301 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      361 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      421 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      481 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      541 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      601 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      661 cgtgtcgtgc ctgagagcgc agctgtgctc ctgggcaccg cgcagtccgc ccccgcggct
      721 cctggccaga ccacccctag gaccccctgc cccaagtcgc agccgttgga tcaggtaagt
      781 atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga
      841 agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc
      901 tccacaggtg atgatcccca taacttcgta taaagtatcc tatacgaagt tatatcaaaa
      961 taggaagacc aatgcttcac catcgacccg aattgccaag catcaccatc gacacataac
     1021 ttcgtataat gtatgctata cgaagttata ctagtgctag cggatccgat atcctaacgt
     1081 ggcttcttct gccaaagcat gatgaggatg ataagggaga tgatggtgag caccaccagg
     1141 gccaggatgg ctgagatcac caccacctta aagggcaagg agtgtggcac cacgatgacc
     1201 tcctgcgtgt cctggcccac agcattcaga tcctcttctg agatgagttt ttgttcgtcg
     1261 acctgcagtt tcagtgcctt gtcattcggt tctttgaaca gtgctttcat ttcgtctgac
     1321 agttcgaaat tcatgttcag ggggttgtta aagttgtact ccagcttgtg ccccaggatg
     1381 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc
     1441 tcgaacttca cctcggcgcg ggtcttgtag gtgccgtcgt ccttgaagct gatggtgcgc
     1501 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg
     1561 gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc
     1621 acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgtt ggtggcatcg
     1681 ccctcgccct cgccgcgcac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc
     1741 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcatgctccc tccggtaccg
     1801 cccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     1861 accatgtgat cgcgcttctc gttggggtct ttgctcagca cggactgggt gctcaggtag
     1921 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     1981 gcgagctgca cgctgccgtc ctccacgttg tggcggatct tgaagttcgc cttgatgccg
     2041 ttcttctgct tgtcggcggt gatatagacg ttgtggctca ccagaattgg atttttgaac
     2101 cacttatcaa accatttttc cgcttcaccg gaggtctgca cctgagcgat ggtgtcatcc
     2161 atcagctttt tgaactgcgg atcatcttta cgcaacatac aaccgtaggc ctcctgagac
     2221 tgcggcttgc cgacgatttc ccagttgtct ggtttcttcg ctttcgcacg ttcaccggcc
     2281 agcagcacgt catccatcat aaaggcaacg gcacgaccgc tttccagggt gcggaaagag
     2341 tcaccgtgat ctttggcgct gatgatgcgc atattcattt tttgctcttc attcagtttg
     2401 ttgagcaaaa cttcagaggt agtgccggaa gtgacgacta cggctttgtc tttcaggttg
     2461 gcaaaatctt tgatatcgcc accctttttg gtcaacaggc gcgtaccgac cacgaaaata
     2521 gtgtcagaga aagccgcctg tttttggcgt tcgacgttgt tggtggtaga accacattca
     2581 aaatcgaaag tgccgttttg cagcagtgga atacggtttt gtgcggtaat cggaatcagt
     2641 tttacctgca agtccggttt gttgagtttc tttttcactg cttcaacaat ggcgttggag
     2701 taatcctgcg agtaacccac cactttttgc tgattgtcgt aataagagaa aggcactgaa
     2761 gattcacggt gaccgacgac aatcacaccg tttttggcga ttttgtccag cgtgctgcct
     2821 gcggcagatc tgtcaccagt ggaacctgga acccagagca gcagtaccca tagcaggagt
     2881 gtgtctgtct ccatggtggc gatatcaccg gtggtaccac tagtataact tcgtatagga
     2941 tactttatac gaagttatca ttgggattct tcctattttg atccaagcat caccatcgac
     3001 cctctagtcc aaatctcacc atcgacccat aacttcgtat agcatacatt atacgaagtt
     3061 atgtccctcg aagaggttcg aaacactgaa aagggccaat tctgcaaatg atccactcga
     3121 gaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3181 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3241 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3301 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3361 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3421 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3481 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     3541 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3601 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3661 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3721 gcatcgatac cgtcgacccg ggcggccgct cgagcatgca tctagagtcg acccgggcgg
     3781 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3841 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3901 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3961 ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     4021 aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta
     4081 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     4141 ggacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat
     4201 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4261 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4321 cggtaatacg gttatccaca gaatcagggg ataacgcagt attctatagt gtcacctaaa
     4381 tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4441 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4501 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4561 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4621 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4681 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4741 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4801 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4861 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4921 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4981 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5041 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5101 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5161 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5221 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5281 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5341 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5401 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5461 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5521 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5581 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5641 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5701 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5761 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5821 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5881 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5941 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6001 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6061 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6121 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6181 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6241 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6301 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6361 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6421 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6481 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6541 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     6601 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     6661 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     6721 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     6781 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     6841 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     6901 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     6961 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     7021 aggct
//
