LOCUS       sequence-204612-        7640 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Medium affinity glutamate sensor ** A newer version of this sensor 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 7640)
  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 7640)
  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/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.A184V.mRuby3
FEATURES             Location/Qualifiers
     source          1..7640
                     /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(2064..2210)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(2214..2243)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     2708..2729
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2832..2853)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(3819..3881)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     regulatory      3878..3887
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3912..3945)
                     /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(4016..4049)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4121..4709
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4174..4194)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4592..4603)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4752..4873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4763..4782)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4817..4836
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5401..5989)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5481..5500)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6160..6951,6952..7020))
                     /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     6783..6802
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7021..7125)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7151..7606)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7288..7309)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7500..7519
                     /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 gcgcgccagg cggggcgggg cggggcgrgg ggcggggcgg 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 caccaagatt ggagtacttg gcaactgcga cttcgcgttg tactacgtac
     1381 gtttcattgt cactttcctc tatcctttcc aggcggtgat caacggcatg gacaccgggc
     1441 atcttgatgt tcccgacggt cttttttgac ctgtaagttg ttacgaaatt gcaatgcaga
     1501 tggccaccac catcaacttt cagtgcaata tcagtgtatc ccctcagacc accatctgct
     1561 ggatacatca tctctgtatt aggctcccaa cccttggtct tcttctgcat cacgggacca
     1621 ttggagggaa agtttacccc tctgactttg acgttgtaaa cgagttcgcc atcctcaagg
     1681 ctggtgtcct gcgtgacggt gacgactcca ccatcttcgt atctcgtaac tctttcccaa
     1741 gtaaaaccct caggaaagga ctgtttaaag aaatcaggaa tgtctgccgg gtacttgata
     1801 aaagtacggc tgccatacat gaacgacgtg gcaagaatgt caaaggcaaa tggcaggggt
     1861 cctccctcga tgactttgat cctcatggtt tgcactccct cgtacggcct gccttctcct
     1921 tcacctgtgc atttgaattg gtggccgttg accgaacctt ccatgaccac cttcatacgc
     1981 atattttcct tgatcagctc ttcgccctta gacaccatac gctcctgcgt gtcctggccc
     2041 acagcatttc cggagcctct agaacgtggc ttcttctgcc aaagcatgat gaggatgata
     2101 agggagatga tggtgagcac caccagggcc aggatggctg agatcaccac caccttaaag
     2161 ggcaaggagt gtggcaccac gatgacctcc tgcgtgtcct ggcccacagc attcagatcc
     2221 tcttctgaga tgagtttttg ttcgtcgacc tgcagtttca gtgccttgtc attcggttct
     2281 ttgaacagtg ctttcatttc gtctgacagt tcgaaattca tgttcagggg gttgttaaag
     2341 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     2401 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtaggtg
     2461 ccgtcgtcct tgaagctgat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     2521 aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc
     2581 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc
     2641 agggtcagct tgccgttggt ggcatcgccc tcgccctcgc cgcgcacgct gaacttgtgg
     2701 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     2761 cccttgctca tgctccctcc ggtaccgccc ttgtacagct cgtccatgcc gagagtgatc
     2821 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     2881 ctcagcacgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     2941 atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc cacgttgtgg
     3001 cggatcttga agttcgcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg
     3061 tggctcacca gaattggatt tttgaaccac ttatcaaacc atttttccgc ttcaccggag
     3121 gtctgcacct gagcgatggt gtcatccatc agctttttga actgcggatc atctttacgc
     3181 aacatacaac cgtaggcctc ctgagactgc ggcttgccga cgatttccca gttgtctggt
     3241 ttcttcgctt tcgcacgttc accggccagc agcacgtcat ccatcataaa ggcaacggca
     3301 cgaccgcttt ccagggtgcg gaaagagtca ccgtgatctt tggcgctgat gatgcgcata
     3361 ttcatttttt gctcttcatt cagtttgttg agcaaaactt cagaggtagt gccggaagtg
     3421 acgactacgg ctttgtcttt caggttggca aaatctttga tatcgccacc ctttttggtc
     3481 aacaggcgcg taccgaccac gaaaatagtg tcagagaaag ccgcctgttt ttggcgttcg
     3541 acgttgttgg tggtagaacc acattcaaaa tcgaaagtgc cgttttgcag cagtggaata
     3601 cggttttgtg aggtaatcgg aatcagtttt acctgcaagt ccggtttgtt gagtttcttt
     3661 ttcactgctt caacaatggc gttggagtaa tcctgcgagt aacccaccac tttttgctga
     3721 ttgtcgtaat aagagaaagg cactgaagat tcacggtgac cgacgacaat cacaccgttt
     3781 ttggcgattt tgtccagcgt gctgcctgcg gcagatctgt caccagtgga acctggaacc
     3841 cagagcagca gtacccatag caggagtgtg tctgtctcca tggtggcgat atcaccggtg
     3901 gtaccactag tataacttcg tataggatac tttatacgaa gttatcattg ggattcttcc
     3961 tattttgatc caagcatcac catcgaccct ctagtccaaa tctcaccatc gacccataac
     4021 ttcgtatagc atacattata cgaagttatg tccctcgaag aggttcgaaa cactgaaaag
     4081 ggccaattct gcaaatgatc cactcgagaa gcttatcgat aatcaacctc tggattacaa
     4141 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     4201 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4261 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     4321 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     4381 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     4441 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     4501 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     4561 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4621 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4681 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacccgggc ggccgcttcg
     4741 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4801 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4861 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     4921 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct
     4981 tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac
     5041 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgccc
     5101 gggctktgcy ckgrsgscgy cmgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     5161 gcagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5221 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5281 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5341 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5401 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5461 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5521 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5581 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5641 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5701 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5761 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5821 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5881 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5941 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6001 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6061 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6121 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6181 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6241 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6301 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6361 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6421 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6481 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6541 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6601 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6661 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6721 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6781 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6841 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6901 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6961 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7021 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7081 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7141 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     7201 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     7261 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     7321 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     7381 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     7441 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     7501 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     7561 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     7621 aggctgcgca actgttggga
//
