LOCUS       sequence-374401-        7475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Glutamate sensor for localization to membrane contatct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7475)
  AUTHORS   Shindo Y, Ashida K, Masamoto K, Takuwa H, Takahashi M, Higuchi M, 
            Ide R, Hotta K, Oka K
  TITLE     Genetically encoded sensors for analysing neurotransmission among 
            synaptically-connected neurons
  JOURNAL   bioRxiv 2022
REFERENCE   2  (bases 1 to 7475)
  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"; journalName: "bioRxiv 
            2022"
COMMENT     Sequence Label: pAAV.hSynapsin.mCherry-P2A-Post-SPHERE-SF-iGlu...
FEATURES             Location/Qualifiers
     source          1..7475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   25..154
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        365..812
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      821..830
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             827..1534
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(972..990)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1240..1259
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1541..1582
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1592..1648
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     2896..2917
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(3254..3265)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    3632..4220
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3685..3705)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4103..4114)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4240..4716
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4439..4458)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4762..4902
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4762..4891
                     /label=AAV2 ITR
     rep_origin      4977..5432
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5064..5083)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5274..5295
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5449..5468)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5568..5590
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5628..5646)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5714..5818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5819..5887,5888..6679)
                     /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(6037..6056)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6850..7438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7339..7358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg
       61 ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
      121 gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgttg caaagatgga
      181 taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag
      241 tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
      301 agttgggggg aggggtcggc agcaaatggt taattaatct agactgcaga gggccctgcg
      361 tatgagtgca agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga
      421 ccgaccccga cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat
      481 cagagagggg gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac
      541 cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag
      601 gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg
      661 tccgcgccgc cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg
      721 ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca
      781 gcggaggagt cgtgtcgtgc ctgagagcgc agtcgagaac gccaccatgg tgagcaaggg
      841 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
      901 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
      961 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     1021 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     1081 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     1141 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     1201 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     1261 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     1321 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     1381 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     1441 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     1501 ccactccacc ggcggcatgg acgagctgta caagggatcc ggcaagccta tccctaatcc
     1561 tctgctggga ctggactcta caggaagcgg agctactaac ttcagcctgc tgaagcaggc
     1621 tggagacgtg gaggagaacc ctggacctga gaccgacacc ctcctgctat gggtgctgct
     1681 gctctgggtg cccggcagca ccggcgacgc accggtcgga ggcaacgacc ctaacctgtt
     1741 cgtcgccctg tacgacttcg tcgcctctgg cgacaacacc ctgagcatca ccaagggcga
     1801 gaagctgaga gtcctgggct acaaccacaa cggcgagtgg tgcgaggccc agaccaagaa
     1861 cggccagggc tgggtcccta gcaactacat cacacctgtc aacagcaccg gtggaggcag
     1921 cggaggaggc tccggagccg caggcagcac gctggacaaa atcgccaaaa acggtgtgat
     1981 tgtcgtcggt caccgtgaat cttcagtgcc tttctcttat tacgacaatc agcaaaaagt
     2041 ggtgggttac tcgcaggatt actccaacgc cattgttgaa gcagtgaaaa agaaactcaa
     2101 caaaccggac ttgcaggtaa aactgattcc gattacctca caaaaccgta ttccactgct
     2161 gcaaaacggc actttcgatt ttgaatgtgg ttctaccacc aacaacgtcg aacgccaaaa
     2221 acaggcggct ttctctgaca ctattttcgt ggtcggtacg cgcctgttga ccaaaaaggg
     2281 tggcgatatc aaagattttg ccaacctgaa agacaaagcc gtagtcgtca cttccggcac
     2341 tacctctgaa gttttgctca acaaactgaa tgaagagcaa aaaatgaata tgcgcatcat
     2401 cagcgccaaa gatcacggtg actctttccg caccctggaa agcggtcgtg ccgttgcctt
     2461 tatgatggat gacgtgctgc tggccggtga acgtgcgaaa gcgaagaaac cagacaactg
     2521 ggaaatcgtc ggcaagccgc agtctcagga ggcctacggt tgtatgttgc gtaaagatga
     2581 tccgcagttc aaaaagctga tggatgacac catcgctcag gtgcagacct ccggtgaagc
     2641 ggaaaaatgg tttgataagt ggttcaaaaa tccaattctg gtgagccaca acgtctatat
     2701 caccgccgac aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacgtgga
     2761 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2821 cgtgctgctg cccgacaacc actacctgag cacccagtcc gtgctgagca aagaccccaa
     2881 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     2941 catggacgag ctgtacaagg gaggaggctc cggaggcgga agcggtaccc tcgagctggt
     3001 gccccacttg cataacctca atgacatcag ccaatacaca agtacaacca ccaaggtccc
     3061 aagcactgac atcacccttc gaccaactcg gaaaaactca acccctgtga ccagcgcatt
     3121 tccaaccgct aagcaagacg accccaaaca acagcctagt ccatttagcg tagaccaaag
     3181 ggactacagc acagagctgt ccgtgaccat agctgtagga gcttccctgc ttttcctgaa
     3241 cattctggct ttcgcggccc tctattacaa gaaggacaaa aggcgccacg acgtacatag
     3301 gagatgtagt ccgcaacgaa ctacaaccaa tgatcttacg cacgcacctg aggaggagat
     3361 catgagtctt cagatgaaac acacagacct ggaccacgag tgtgaatcca tccatcccca
     3421 tgaggtggtg ctgaggacag catgccctcc ggactatacc ctggcaatgc gccgaagccc
     3481 agacgacatt cccctgatga cacccaatac cataaccatg attcccaaca ctattccagg
     3541 aatccagcct ctccacactt tcaacacatt cactggtgga cagaataata ctctccctca
     3601 tccgcatcca cacccccact cccacagcta aaatcaacct ctggattaca aaatttgtga
     3661 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3721 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3781 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3841 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3901 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3961 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     4021 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     4081 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     4141 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     4201 cctttgggcc gcctccccgc atcgataccg gctcgagacg ggtggcatcc ctgtgacccc
     4261 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     4321 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     4381 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     4441 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     4501 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     4561 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     4621 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     4681 ttacaggcgt gaaccactgc tcccttccct gtccttctcg agctgatttt gtaggtaacc
     4741 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     4801 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4861 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     4921 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     4981 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5041 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5101 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5161 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     5221 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5281 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     5341 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5401 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     5461 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5521 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5581 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     5641 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5701 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5761 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5821 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5881 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5941 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6001 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6061 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6121 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6181 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6241 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6301 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6361 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6421 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6481 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6541 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6601 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6661 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6721 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6781 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6841 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6901 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6961 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7021 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7081 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7141 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7201 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7261 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7321 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7381 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7441 cgccagcaac gcggcctttt tacggttcct ggcct
//
