LOCUS       sequence-369252-        7468 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 7468)
  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 7468)
  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: Ttx-3-mCherry-SL2-Post-SPHERE-SF-iGluSnFR
FEATURES             Location/Qualifiers
     source          1..7468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    29..53
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(897..921)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             953..1663
                     /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(1098..1116)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1366..1385
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4923..4945)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5045..5064
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5142..5160)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5228..5332
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5333..5401,5402..6193)
                     /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(5551..5570)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6364..6952
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6853..6872
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7106..7123
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7240..7261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7276..7281,7282..7299,7300..7306)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7314..7330
                     /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     7319..7341
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7338..7354
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7338..7354
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 acttggaaat gaaataagct acccatcaac aagtttgtac aaaaaagcag gctggatcca
       61 agcttttttg aaacgatctt tgttatcaat gatgtgtcat aaaaacacat gttttccagt
      121 tcaatagtga acacgtaaaa accagaagaa tgtttgattt ggaagacgta gcagtatttt
      181 tttgaaaaca ttttttttaa tgaaataatt taatatatta aaattgtgtt ttttttatga
      241 cccgattata tcacttatct attcagacgt tgttaaattt tccctaaact tttttttact
      301 tcagacaatt gctgctgatg cttaccaact cttcctcaaa ggagactatt gccacccagg
      361 ttgttacaaa ctcggtgcct tctgcattcc agtttgattg gagctcacat ttcaaagaag
      421 aatatgattt ggtggaaaag ttgaactccc cgcaatatac caacttaacg tccatatccc
      481 cttcctcatc aactgattcc aacaacttaa ttctacggca aattcagata ccgaagattg
      541 agccggcact tttgagtgag catctattac agcgattttc tctaatttaa cccgaaatta
      601 taacctgttt ttttttcgaa aatgactcat ttttctcgaa tacagagttg agaataaaaa
      661 tagcaaggtc gtgtatttta tcatgtcgct atttttctta acgaagccaa taacaatgcc
      721 acttatttca agtcttgaga aacaaattat ttgacgaaaa tgtttttttt tcaaagattt
      781 actgctttaa ctttacagat cagtgttgtc tctgcacatt tgcaattgtc gataaagaaa
      841 tcagtgttgt tgacggaaaa tattatcata ataattgtct tcggtgtcaa aggatcaccc
      901 agctttcttg tacaaagtgg ttgatgggac gatatcgggt accggtagaa aaatggtgag
      961 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     1021 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     1081 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     1141 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     1201 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     1261 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     1321 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     1381 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     1441 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     1501 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     1561 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     1621 gggccgccac tccaccggcg gcatggacga gctgtacaag taagctgtct catcctactt
     1681 tcacctagtt aactgcttgt cttaaaatct atgcttctct ttagtatcta aaattttcct
     1741 agaagcttac aagtatataa atggtctctt ctcaataaag gttgtatatt tattcatctt
     1801 attgaatctg ccatttcctc gtttttgcga gtttatatac cttccaattt tctttctatt
     1861 gtattttcaa cttctaattt taattcaggg aaactgctgt accggtagaa aaaatggaga
     1921 ccgacaccct tcttctttgg gtccttcttc tttgggttcc aggatctacc ggagatgctc
     1981 cagttggagg aaacgatcca aaccttttcg tcgctcttta cgatttcgtc gcttccggag
     2041 ataacaccct ttctatcact aagggagaga agcttcgtgt tcttggatac aaccataacg
     2101 gagaatggtg cgaggctcaa actaagaacg gacaaggatg ggttccatct aactacatca
     2161 ctccagttaa ctctactgga ggaggatctg gaggaggatc tggagctgct ggatccaccc
     2221 ttgataagat cgctaagaac ggagttatcg ttgttggaca ccgtgagtcc tctgttccat
     2281 tctcctacta cgataaccaa caaaaggttg ttggatactc tcaagactac tccaacgcta
     2341 tcgtcgaggc tgttaagaag aagcttaata agccagatct tcaagttaag cttatcccaa
     2401 tcacttctca aaaccgtatc ccacttcttc aaaacggaac cttcgacttc gagtgcggat
     2461 ccactaccaa caacgttgag cgtcaaaagc aagctgcctt ctccgatacc atcttcgttg
     2521 ttggaacccg tcttcttact aagaagggag gagacattaa ggacttcgct aaccttaagg
     2581 taagtttaaa catatatata ctaactaacc ctgattattt aaattttcag gataaggctg
     2641 ttgttgttac ctctggaacc acctccgagg tccttcttaa caagcttaac gaggaacaga
     2701 agatgaacat gcgtattatc tccgctaagg accacggaga ttccttccgt acccttgagt
     2761 ctggacgtgc tgtcgctttc atgatggacg acgtccttct tgctggagag cgtgctaagg
     2821 ctaagaagcc agataactgg gagatcgttg gaaagccaca atcccaagaa gcttacggat
     2881 gcatgcttcg caaggacgac ccacaattca agaagcttat ggacgatacc atcgcccaag
     2941 ttcaaacctc tggagaggct gagaagtggt tcgacaaatg gttcaagaac ccaatccttg
     3001 tttctcacaa cgtttacatc accgctgata agcaaaagaa cggaattaag gctaacttca
     3061 agatccgtca taacgttgag gatggatccg tccaacttgc tgatcattac caacaaaaca
     3121 ctccaatcgg agatggacca gttcttcttc cagataacca ctacctttct acccaatccg
     3181 ttctttctaa ggatccaaac gagaagcgtg accacatggt ccttcttgag ttcgtcaccg
     3241 ctgctggaat cacccttgga atggatgaac tttataaagg tggaggatct ggaggaggat
     3301 ctggaaccct tgagcttgtc ccacaccttc acaaccttaa cgatatctct caatacactt
     3361 ctaccaccac caaggtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt
     3421 ttcaggttcc atctaccgat atcaccctcc gcccaacccg caagaactct accccagtta
     3481 cctccgcttt cccaaccgcc aagcaagatg atccaaagca gcaaccatct ccattctccg
     3541 ttgatcaacg tgattactct accgagcttt ccgttaccat cgccgtcgga gcctcccttc
     3601 tcttccttaa catccttgcc ttcgctgctc tttactacaa gaaggataag cgccgtcacg
     3661 atgttcaccg ccgctgctct ccacaacgca ccaccaccaa cgacctcact cacgctccag
     3721 aagaggagat catgtctctc caaatgaagc acaccgatct tgatcacgaa tgcgagtcta
     3781 tccacccaca cgaggttgtt cttcgtaccg cttgcccacc agattacacc cttgctatgc
     3841 gtcgttcccc agatgatatc ccacttatga ctccaaacac tatcactatg atcccaaaca
     3901 ccatcccagg aatccaacca cttcatacct tcaacacttt caccggagga caaaacaaca
     3961 cccttccaca cccacaccca cacccacact ctcactctta atctagacaa ttccaactga
     4021 gcgccggtcg ctaccattac caacttgtct ggtgtcaaaa ataatagggg ccgctgtcat
     4081 cagagtaagt ttaaactgag ttctactaac taacgagtaa tatttaaatt ttcagcatct
     4141 cgcgcccgtg cctctgactt ctaagtccaa ttactcttca acatccctac atgctctttc
     4201 tccctgtgct cccaccccct atttttgtta ttatcaaaaa acttctctta atttctttgt
     4261 tttttagctt cttttaagtc acctctaaca atgaaattgt gtagattcaa aaatagaatt
     4321 aattcgtaat aaaaagtcga aaaaaattgt gctccctccc cccattaata ataattctat
     4381 cccaaaatct acacaatgtt ctgtgtacac ttcttatgtt ttttacttct gataaatttt
     4441 tttgaaacat catagaaaaa accgcacaca aaatacctta tcatatgtta cgtttcagtt
     4501 tatgaccgca atttttattt cttcgcacgt ctgggcctct catgacgtca aatcatgctc
     4561 atcgtgaaaa agttttggag tatttttgga atttttcaat caagtgaaag tttatgaaat
     4621 taattttcct gcttttgctt tttggggttt cccctattgt ttgtcaagat ttcgaggacg
     4681 gcgtttttct tgctaaaatc acaagtattg atgagcacga tgcaagaaag atcggaagaa
     4741 ggtttgggtt tgaggctcag tggaaggtga gtagaagttg ataatttgaa agtggagtag
     4801 tgtctatggg gtttttgcct taaatgacag aatacattcc caatatacca aacataactg
     4861 tttcctacta gtcggccgta cgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     4921 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     4981 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     5041 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     5101 agatgcgtaa ggagaaaata ccgcatcagg cggccttaag ggcctcgtga tacgcctatt
     5161 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5221 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5281 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5341 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5401 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5461 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5521 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5581 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5641 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5701 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5761 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5821 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5881 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5941 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6001 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6061 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6121 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6181 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6241 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6301 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6361 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6421 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6481 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6541 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6601 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6661 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6721 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6781 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6841 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6901 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6961 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7021 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7081 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     7141 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     7201 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     7261 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     7321 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tgtaagttta
     7381 aacatgatct tactaactaa ctattctcat ttaaattttc agagcttaaa aatggctgaa
     7441 atcactcaca acgatggata cgctaaca
//
