LOCUS       sequence-486617-        9667 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 9667)
  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 9667)
  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: Str-2-TagBFP-SL2-Pre-SPHERE-SF-iGluSnFR
FEATURES             Location/Qualifiers
     source          1..9667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    14..38
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3747..3771)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7107..7129)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7229..7248
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7326..7344)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7412..7516
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7517..7585,7586..8377)
                     /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(7735..7754)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8548..9136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9037..9056
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9290..9307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9424..9445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9460..9465,9466..9483,9484..9490)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9498..9514
                     /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     9503..9525
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9522..9538
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9522..9538
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagctaccca tcaacaagtt tgtacaaaaa agcaggctaa gcgtcaagag acaggataca
       61 actgataata caaaatgaat gtaagagttt ctccactgaa ttcccatttc attacacttt
      121 taattagtaa gcactcaccc ctggccagcc tcctgaataa tatagataaa ctgaaccaaa
      181 tgggaacggg tgtcccatac tcctacagta accaacggcg ggaatcataa aagcggtaca
      241 tccgaatggt ataagaaaca acaaaagtaa taaaatactc attaattttg ggcagatctg
      301 aaaaaatagt acatgtatac atatattccc atgtgaacta atacttcctt acaaaaaagt
      361 gattattggg acgtaacaag attatcactc tgaacagtga caagtagaat ggaagaaaaa
      421 agacagaata gttaaaatat taaaaattaa gaaaatagtt tttataaaat gatttggttc
      481 gacaaacgca caccatgaag tgattattcc agttgcagga atagagccac gaagaaactc
      541 aaacatgaaa aacgaaatac tcagtaaatt gataggaaac gtttctaaat ctttcgcttc
      601 ttgagtttaa atatttcgtt gcataaatca actcacacag aaaaatagca tcgcaatgca
      661 gatgaaaata tatttattca ttggagcaat tttggaccat ccaataagtg ctgacaattt
      721 tccacattac agcaatggta ggtggcatat aaatcaatgg gatcaacgcc agaattgtag
      781 gaaacgtaaa agtatactga aaactttggt attcggggat tccgtaaata gaaccgttca
      841 ttttttttag attaccaatt tacataaaat atgaatatca atgatttgag aaagaaaaac
      901 atgacgtaga tgtgggcttg aagtaattgt attatatatt tattatattt tattacatta
      961 ttattttgtg gaccggaagt aaaaattcct tcatgaaaac catttatgag attgaaaaat
     1021 aacgtttcca aaaaacaagg ctaaaaatat ttaattgatg agaaaaatct atatgaaaaa
     1081 ataaaaaaat gaaaaattta taaagtatca aaaaaaattt agattaacat ttacgtaatt
     1141 atttttagcc ttggattctc atcacattat ttaatttaat ttaatttaat ttgtttacgc
     1201 tcgcgaggag acgtggaaaa agttgaaaaa aaaagaaaaa tgttcacata caatagatca
     1261 taaatacaat acactgaata agtgggaaga aaactaagaa aaataaatag gaacgaaagt
     1321 ttgttaacag tttgctcctt taccttgcat tgcttatctc actctccacc gcttgatttt
     1381 gctccgaatt ccaccacatg cgcgcttctg aacggtctgt gggctctgac acccacctgg
     1441 tcgagttgac cgttcaacat ggtgcatcga ccgggaatat tgctcgtgca atcgccaaac
     1501 tatgtcttcg gatcgtaaat tcgttcgtaa aaatcgatcg taaaaaactc tccatctcca
     1561 attccaggag ctgacggttc taatgaaacc gaaagttcag ggaaacgcga gactctacgg
     1621 actcgcaaga gtcaaaagtc tacaagggga actcgtgcag ccacaccagg ccgcttgaaa
     1681 agcttcgctg ccccaattaa agggtacatt acccacgcaa ggttggtcag gcaggaggaa
     1741 ataatgttat gtgggtagtt tatgttgcaa tcatcagttg tcagacatct cgttattcaa
     1801 gatctgatat atgtttaaga aacttcgggt tactcagcgt actacaataa aatgctttct
     1861 agtttcagtt ttacatctgc tccatgagac tttctttttg gaacaagttt tggaacgagt
     1921 catattattt tttcagaatg aagcgttctt cgccattgac tgctccgtaa tagatttgaa
     1981 gtcgtgtttt acctatatag gcatgaggca ggcggagttg aaacaattga ttatacactc
     2041 atacagtttt atcttggttc aaatttttgc ccaatataaa atgtgcagct tattgatcat
     2101 attaagacaa aaaagtgttt aaaaaaagta actccaatta ggtattatgt atgcaagttt
     2161 tcaaagaagc caaccaaaca gtgatgtgcg ttatttgaat taaatcttat cttcccagat
     2221 cagttttctc gaattcatag tttttaattt ttataaattg atatttcagc atttcgtaat
     2281 tctcatgttc taaataggaa acatctggga tgcgttgtag acatgttggg acaacgtcga
     2341 atatttttct caaattccat ttctatttcg ttttaggcaa agtgaaacaa aaaattttca
     2401 ataccaaata tcagtctaag taattcgaaa aacggtattt tatactccca aaaggctacg
     2461 acgcctataa aacaaaactg accagacttc cacacctggg agagataggg tcagtttttg
     2521 ttcgtttgtc tcattggatt tcctcgctga tacatgattt aaaacatact tttattgtat
     2581 agaacgataa tttttgtgtg gtttattatg aagcaaacat cattcataca tcaaatgcgt
     2641 gttttcagtt cagcttcctt tgttcaccaa taaagttttt aattccttga aagggatgga
     2701 ttatcctttt atcgaaaaac gtagaacttc gtgtacgttt taaaataatc tttcaattca
     2761 agctgtttct tttattaata ttacattttt ttagttgatt tcacttaaaa aactctattc
     2821 attcggatta ttggttgaag gaactctgat ttggttgaaa tatcaaattt aaacataata
     2881 ctgttaatgg ttcgattata ccaagaaaat agttcaaatc ttctctcaat aataatcgta
     2941 gacattgttt tcaatagtat atagtctgtc tctaattttc aagctcttta gagaattaaa
     3001 catattgttt ttgtatttca gttttattaa tccctgtttg ttctcaacaa tttcttatct
     3061 agatcagtta ttcagttatg tagatatgct tgagattctg aaatcggaaa gtataatcat
     3121 aaacgaaatt ttggatatta tttgagcttt taaatttttt tttcaacatt agggtaacat
     3181 ttttagtcga tctttgtcca ttttctgaga acctactatg ttttttgttt gagcttaaaa
     3241 attcgattga acaactgtac agtatagcgt cacttcttag gattcctgat attttatgct
     3301 gaactactgg ttcataaaat tttctttcaa ataaaactgt ctgatttgtt tattacacaa
     3361 cgaagtttgt atgcgtaaac gataaaaaca tcttctagac agaccaaagt ctttcttcaa
     3421 tagtcaaatc atttcaaatt gttcctaatg atcctttcat gaatatttgt acaaaataac
     3481 agttgacaaa ccaaatattt tgtgaaaaat tggagtgatt agcgaatccc gtaggaaaaa
     3541 ccaggagtca gttaagtgaa gcattttctt gcaagaaaga aataaatgat ttcatgttcc
     3601 caactggtga taatatctct ctaaggagtt atcatgtgtt gtttttggaa attaatattc
     3661 ttcacttttt gtgtttttat ttcaaccatt ctcgattata attttgcttc cagacttttt
     3721 tgtccgaata ctcgtgatcc ataaaaaccc agctttcttg tacaaagtgg ttgatgggac
     3781 gatatcgggt accggtagaa aaatgtctga acttatcaag gaaaacatgc atatgaagct
     3841 ttacatggaa ggaaccgttg ataaccacca cttcaagtgc acttctgaag gagagggaaa
     3901 gccatacgaa ggaactcaaa ccatgcgcat caaggttgtt gaaggaggac cacttccatt
     3961 cgctttcgac atccttgcca cctctttcct ttacggatct aaggtaagtt taaacatata
     4021 tatactaact aaccctgatt atttaaattt tcagactttc atcaaccata ctcaaggaat
     4081 tccagatttc ttcaagcaat ctttcccaga aggattcact tgggagcgcg ttactaccta
     4141 cgaggatgga ggagttctta ccgccaccca agatacctcc cttcaagatg gatgccttat
     4201 ctacaacgtt aagatccgcg gagttaactt cacttctaac ggaccagtta tgcaaaagaa
     4261 gactcttgga tgggaggctt tcactgagac cctttaccca gccgatggag gacttgaggg
     4321 acgcaacgac atggctctta agcttgttgg aggatcccac cttatcgcta acatcaaggt
     4381 aagtttaaac agttcggtac taactaacca tacatattta aattttcaga ctacctaccg
     4441 ctctaagaag ccagccaaga atcttaagat gccaggagtt tattacgtcg attaccgcct
     4501 tgagcgcatc aaggaagcta acaacgagac ttacgttgag caacacgagg ttgctgtcgc
     4561 tcgttactgc gatcttccat ctaagcttgg acacaagctc aactaagctg tctcatccta
     4621 ctttcaccta gttaactgct tgtcttaaaa tctatgcttc tctttagtat ctaaaatttt
     4681 cctagaagct tacaagtata taaatggtct cttctcaata aaggttgtat atttattcat
     4741 cttattgaat ctgccatttc ctcgtttttg cgagtttata taccttccaa ttttctttct
     4801 attgtatttt caacttctaa ttttaattca gggaaactgc tgtaccggta gaaaaaatgg
     4861 agaccgacac ccttcttctt tgggtccttc ttctttgggt tccaggatct accggagatg
     4921 ctccagttgg aggaatgtct aagggagaag agcttttcac cggagttgtt ccaatccttg
     4981 tcgagcttga tggagatgtt aacggacaca agttctctgt tcgtggagag ggagagggag
     5041 acgctaccaa cggaaagctt actcttaagt tcatctgcac tactggaaag cttccagttc
     5101 catggccaac ccttgttacc acccttactt acggagttca atgcttctct cgttacccag
     5161 atcatatgaa gcaacatgat ttcttcaagt ctgctatgcc agagggatac gttcaagagc
     5221 gtaccatctc tttcaaggat gatggaacct acaaggtaag tttaaacata tatatactaa
     5281 ctaaccctga ttatttaaat tttcagaccc gtgctgaggt taagttcgag ggagataccc
     5341 ttgttaaccg tatcgagctt aagggaatcg atttcaagga agatggaaac atccttggac
     5401 ataagcttga atacaatttc aataatccac ttaacatgaa cttcgagctt tccgacgaga
     5461 tgaaggctct tttcaaggag ccaaacgata aggctcttaa gactggagga tccggaggat
     5521 ctggaggatc ccgtgcccca acttattccc caccaccacc accaggtggt ggatctggag
     5581 gaggatctgg aactgaggtt ccatcttcca tgaccaccga gtctaccgct accgctatgc
     5641 aatctgagat gtctacttct atcatggaga ctactaccac ccttgccacc tctaccgccc
     5701 gccgcggaaa gccaccaact aaggtaagtt taaacagttc ggtactaact aaccatacat
     5761 atttaaattt tcaggaacca atctctcaaa ccaccgacga catcctcgtc gcttccgctg
     5821 agtgcccatc cgatgacgag gatatcgatc catgcgagcc atcttctgga ggacttgcta
     5881 acccaacccg cgttggagga cgtgagccat acccaggatc tgctgaggtt atccgtgagt
     5941 cctcttccac caccggaatg gttgttggaa tcgtcgctgc tgccgccctt tgcatcctta
     6001 tccttcttta cgctatgtac aagtaccgca accgcgacga gggatcctac cacgttgacg
     6061 agtctcgtaa ctacatctct aactctgctc aatccaacgg agccgttgtt aaggagaagc
     6121 aaccatcttc tgctaagtct gctaataaga ataagaagaa caaggacaag gaatactacg
     6181 tctaatctag acaattccaa ctgagcgccg gtcgctacca ttaccaactt gtctggtgtc
     6241 aaaaataata ggggccgctg tcatcagagt aagtttaaac tgagttctac taactaacga
     6301 gtaatattta aattttcagc atctcgcgcc cgtgcctctg acttctaagt ccaattactc
     6361 ttcaacatcc ctacatgctc tttctccctg tgctcccacc ccctattttt gttattatca
     6421 aaaaacttct cttaatttct ttgtttttta gcttctttta agtcacctct aacaatgaaa
     6481 ttgtgtagat tcaaaaatag aattaattcg taataaaaag tcgaaaaaaa ttgtgctccc
     6541 tccccccatt aataataatt ctatcccaaa atctacacaa tgttctgtgt acacttctta
     6601 tgttttttac ttctgataaa tttttttgaa acatcataga aaaaaccgca cacaaaatac
     6661 cttatcatat gttacgtttc agtttatgac cgcaattttt atttcttcgc acgtctgggc
     6721 ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt ggagtatttt tggaattttt
     6781 caatcaagtg aaagtttatg aaattaattt tcctgctttt gctttttggg gtttccccta
     6841 ttgtttgtca agatttcgag gacggcgttt ttcttgctaa aatcacaagt attgatgagc
     6901 acgatgcaag aaagatcgga agaaggtttg ggtttgaggc tcagtggaag gtgagtagaa
     6961 gttgataatt tgaaagtgga gtagtgtcta tggggttttt gccttaaatg acagaataca
     7021 ttcccaatat accaaacata actgtttcct actagtcggc cgtacgggcc ctttcgtctc
     7081 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     7141 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     7201 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     7261 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcggcct
     7321 taagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     7381 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     7441 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     7501 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     7561 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     7621 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     7681 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     7741 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     7801 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     7861 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7921 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7981 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     8041 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     8101 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     8161 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     8221 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     8281 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     8341 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     8401 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     8461 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     8521 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     8581 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     8641 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     8701 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     8761 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     8821 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     8881 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     8941 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     9001 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     9061 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     9121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     9181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     9241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     9301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     9361 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     9421 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     9481 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     9541 tgattacgcc aagctgtaag tttaaacatg atcttactaa ctaactattc tcatttaaat
     9601 tttcagagct taaaaatggc tgaaatcact cacaacgatg gatacgctaa caacttggaa
     9661 atgaaat
//
