LOCUS       sequence-350412-        6504 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Fluorescent reporter for glutamate, third generation, variant 857 in
            GPI backbone. iGluSnFR3.v857.GPI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6504)
  AUTHORS   Aggarwal A, Liu R, Chen Y, Ralowicz AJ, Bergerson SJ, Tomaska F, 
            Mohar B, Hanson TL, Hasseman JP, Reep D, Tsegaye G, Yao P, Ji X, 
            Kloos M, Walpita D, Patel R, Mohr MA, Tillberg PW, Looger LL, Marvin
            JS, Hoppa MB, Konnerth A, Kleinfeld D, Schreiter ER, Podgorski K
  TITLE     Glutamate indicators with improved activation kinetics and 
            localization for imaging synaptic transmission.
  JOURNAL   Nat Methods. 2023 May 4. doi: 10.1038/s41592-023-01863-6.
  PUBMED    37142767
REFERENCE   2  (bases 1 to 6504)
  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"; journalName: "Nat 
            Methods. 2023 May 4. doi: 10.1038/s41592-023-01863-6."
COMMENT     Sequence Label: pAAV.CAG.iGluSnFR3.v857.GPI
FEATURES             Location/Qualifiers
     source          1..6504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     390..412
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     474..493
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      527..536
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     533..595
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     CDS             2177..2206
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    2339..2927
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2392..2412)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2810..2821)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2970..3091
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2981..3000)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3035..3054
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3253..3382
                     /label=AAV2 ITR
     rep_origin      complement(3619..4207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3699..3718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4378..5169,5170..5238))
                     /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     5001..5020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5239..5343)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5369..5824)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5506..5527)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5718..5737
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5896..6025
                     /label=AAV2 ITR
     promoter        join(6338..6504,1..111)
                     /label=chicken beta-actin promoter
ORIGIN
        1 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
       61 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
      121 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
      181 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
      241 aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag
      301 gggctccggg agggcccttt gtgcgggggg agcggctcgg ggctgtccgc ggggggacgg
      361 ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc
      421 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
      481 gctggttatt gtgctgtctc atcattttgg caaagaattg gatcccgcca ccatggagac
      541 agacacactc ctgctatggg tactgctgct ctgggttcca ggttccactg gtgacagatc
      601 tgggtccgcc gccggaagta ctcttgataa gatcgctaaa aatggtgtaa ttgttgtcgg
      661 ccaccgagac tcttctgtgc cattcagcta ttatgattct caacacaaag tggtgggcta
      721 ttcacaagag tattctaacg ccattgttga agcagttaag aagaagttga ataaaccaga
      781 ccttcaagta aaactcattc ctataacatc ccaaaaccgc atacctctct tgcaaaacgg
      841 cacttacgat ttcgagtgtg ggagcactac taataatgtc gaacgccaga aacaagccgc
      901 attctccgac accatcttcg tggtgggcac tcgactgttg acaaagaaag gcggtgatat
      961 taaggatttc gcaaatctga aggataaggc agtcgtggtc accagcggga ccacctcaga
     1021 agtactcctt aataaactca atgaagaaca aaagatgaat atgcgaatca tctctgctaa
     1081 ggaccacggt gactcttttc gcacattgga aagtgggcgc gccgtagcat ttatgatgga
     1141 cgactctctg ctcgctggcg agcgcgccaa agccaaaaaa ccagacgact gggaaattgt
     1201 aggtaaacct caaagccaag aggcatatgg ctgcatgttg agaaaagatg acccccaatt
     1261 caagaaactg atggacgaca ctatagctca ggtccagact tcaggggaag ctgagaagtg
     1321 gtttgataaa tggttcaaga acccaatctt ggtaaatcat accgtttata ttaccgctga
     1381 caaacaaaag aatgggataa aggcaaattt caagattcac cataacgtag aggatggaag
     1441 cgtgcagttg gcagaccatt atcaacagaa tacccctatt ggcgacggac ccgtgctcct
     1501 ccctgacaac cactacttgt cctatcaaag tgttctctca aaggacccca atgagaagcg
     1561 agaccacatg gtacttctcg aattcgttac cgccaccggg aattcacttg gcatggatga
     1621 attgtataag gggggtactg gcggcagcat gtccaagggt gaggagcttt ttaccggagt
     1681 agtaccaatt ctcgttgaac ttgacggaga cgtcaacggg cacaagttta gcgtacgagg
     1741 tgagggggag ggtgatgcaa ccaacggtaa acttactctc aaattcatat gcactaccgg
     1801 aaagctgcca gtaccttggc caacactggt cactaccctc tcctatggcg ttcaatgttt
     1861 tgctcgatac cctgaccaca tgaaacagca tgacttcttt aaatcagcca tgcctgaagg
     1921 gtatgtccag gagcggacta tatcatttaa agacgatgga acctataaga ctcgggctga
     1981 agtgaagctc gagggtgaca cattggttaa tcggatcgag cttaagggca tagattttaa
     2041 agaagatggt aatattcttg gccacaaatt ggagtacaat tttaataatc ctttgaacat
     2101 gaactttgag ctctccgatg aaatgaaggc actctttaaa gagccaaacg acaaagctct
     2161 gaaactgcag gtcgacgaac aaaaactcat ctcagaagag gatctgtcag ataacgacta
     2221 cggtaaagat gggttggctg gggcttcagg aggcagacga aatggcgcta taactgtgct
     2281 cagtttcata acgttctacg tggcagcatt tatggtattg ttgtagaagc ttatcgataa
     2341 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2401 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2461 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2521 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2581 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2641 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2701 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     2761 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2821 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     2881 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     2941 acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     3001 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3061 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3121 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3181 taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     3241 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3301 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3361 cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3421 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3481 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3541 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3601 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3661 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3721 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3781 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3841 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3901 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3961 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4021 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4081 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4141 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4201 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4261 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4321 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4381 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4441 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4501 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4561 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4621 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4681 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4741 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4801 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4861 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4921 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4981 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5041 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5101 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5161 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5221 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5281 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5341 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     5401 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     5461 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     5521 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     5581 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     5641 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5701 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5761 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5821 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     5881 tcgctattac gccagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
     5941 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
     6001 gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatct
     6061 acgtagccat gctctaggaa gatcgtacca ttgacgtcaa taatgacgta tgttcccata
     6121 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     6181 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     6241 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     6301 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
     6361 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     6421 ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccagg cggggcgggg
     6481 cggggcgagg ggcggggcgg ggcg
//
