LOCUS       sequence-369254-        9667 bp    DNA     circular SYN 15-APR-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 Apr 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    3..27
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3736..3760)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7096..7118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7218..7237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7315..7333)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7401..7505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7506..7574,7575..8366)
                     /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(7724..7743)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8537..9125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9026..9045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9279..9296
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9413..9434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9449..9454,9455..9472,9473..9479)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9487..9503
                     /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     9492..9514
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9511..9527
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9511..9527
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 caacaagttt gtacaaaaaa gcaggctaag cgtcaagaga caggatacaa ctgataatac
       61 aaaatgaatg taagagtttc tccactgaat tcccatttca ttacactttt aattagtaag
      121 cactcacccc tggccagcct cctgaataat atagataaac tgaaccaaat gggaacgggt
      181 gtcccatact cctacagtaa ccaacggcgg gaatcataaa agcggtacat ccgaatggta
      241 taagaaacaa caaaagtaat aaaatactca ttaattttgg gcagatctga aaaaatagta
      301 catgtataca tatattccca tgtgaactaa tacttcctta caaaaaagtg attattggga
      361 cgtaacaaga ttatcactct gaacagtgac aagtagaatg gaagaaaaaa gacagaatag
      421 ttaaaatatt aaaaattaag aaaatagttt ttataaaatg atttggttcg acaaacgcac
      481 accatgaagt gattattcca gttgcaggaa tagagccacg aagaaactca aacatgaaaa
      541 acgaaatact cagtaaattg ataggaaacg tttctaaatc tttcgcttct tgagtttaaa
      601 tatttcgttg cataaatcaa ctcacacaga aaaatagcat cgcaatgcag atgaaaatat
      661 atttattcat tggagcaatt ttggaccatc caataagtgc tgacaatttt ccacattaca
      721 gcaatggtag gtggcatata aatcaatggg atcaacgcca gaattgtagg aaacgtaaaa
      781 gtatactgaa aactttggta ttcggggatt ccgtaaatag aaccgttcat tttttttaga
      841 ttaccaattt acataaaata tgaatatcaa tgatttgaga aagaaaaaca tgacgtagat
      901 gtgggcttga agtaattgta ttatatattt attatatttt attacattat tattttgtgg
      961 accggaagta aaaattcctt catgaaaacc atttatgaga ttgaaaaata acgtttccaa
     1021 aaaacaaggc taaaaatatt taattgatga gaaaaatcta tatgaaaaaa taaaaaaatg
     1081 aaaaatttat aaagtatcaa aaaaaattta gattaacatt tacgtaatta tttttagcct
     1141 tggattctca tcacattatt taatttaatt taatttaatt tgtttacgct cgcgaggaga
     1201 cgtggaaaaa gttgaaaaaa aaagaaaaat gttcacatac aatagatcat aaatacaata
     1261 cactgaataa gtgggaagaa aactaagaaa aataaatagg aacgaaagtt tgttaacagt
     1321 ttgctccttt accttgcatt gcttatctca ctctccaccg cttgattttg ctccgaattc
     1381 caccacatgc gcgcttctga acggtctgtg ggctctgaca cccacctggt cgagttgacc
     1441 gttcaacatg gtgcatcgac cgggaatatt gctcgtgcaa tcgccaaact atgtcttcgg
     1501 atcgtaaatt cgttcgtaaa aatcgatcgt aaaaaactct ccatctccaa ttccaggagc
     1561 tgacggttct aatgaaaccg aaagttcagg gaaacgcgag actctacgga ctcgcaagag
     1621 tcaaaagtct acaaggggaa ctcgtgcagc cacaccaggc cgcttgaaaa gcttcgctgc
     1681 cccaattaaa gggtacatta cccacgcaag gttggtcagg caggaggaaa taatgttatg
     1741 tgggtagttt atgttgcaat catcagttgt cagacatctc gttattcaag atctgatata
     1801 tgtttaagaa acttcgggtt actcagcgta ctacaataaa atgctttcta gtttcagttt
     1861 tacatctgct ccatgagact ttctttttgg aacaagtttt ggaacgagtc atattatttt
     1921 ttcagaatga agcgttcttc gccattgact gctccgtaat agatttgaag tcgtgtttta
     1981 cctatatagg catgaggcag gcggagttga aacaattgat tatacactca tacagtttta
     2041 tcttggttca aatttttgcc caatataaaa tgtgcagctt attgatcata ttaagacaaa
     2101 aaagtgttta aaaaaagtaa ctccaattag gtattatgta tgcaagtttt caaagaagcc
     2161 aaccaaacag tgatgtgcgt tatttgaatt aaatcttatc ttcccagatc agttttctcg
     2221 aattcatagt ttttaatttt tataaattga tatttcagca tttcgtaatt ctcatgttct
     2281 aaataggaaa catctgggat gcgttgtaga catgttggga caacgtcgaa tatttttctc
     2341 aaattccatt tctatttcgt tttaggcaaa gtgaaacaaa aaattttcaa taccaaatat
     2401 cagtctaagt aattcgaaaa acggtatttt atactcccaa aaggctacga cgcctataaa
     2461 acaaaactga ccagacttcc acacctggga gagatagggt cagtttttgt tcgtttgtct
     2521 cattggattt cctcgctgat acatgattta aaacatactt ttattgtata gaacgataat
     2581 ttttgtgtgg tttattatga agcaaacatc attcatacat caaatgcgtg ttttcagttc
     2641 agcttccttt gttcaccaat aaagttttta attccttgaa agggatggat tatcctttta
     2701 tcgaaaaacg tagaacttcg tgtacgtttt aaaataatct ttcaattcaa gctgtttctt
     2761 ttattaatat tacatttttt tagttgattt cacttaaaaa actctattca ttcggattat
     2821 tggttgaagg aactctgatt tggttgaaat atcaaattta aacataatac tgttaatggt
     2881 tcgattatac caagaaaata gttcaaatct tctctcaata ataatcgtag acattgtttt
     2941 caatagtata tagtctgtct ctaattttca agctctttag agaattaaac atattgtttt
     3001 tgtatttcag ttttattaat ccctgtttgt tctcaacaat ttcttatcta gatcagttat
     3061 tcagttatgt agatatgctt gagattctga aatcggaaag tataatcata aacgaaattt
     3121 tggatattat ttgagctttt aaattttttt ttcaacatta gggtaacatt tttagtcgat
     3181 ctttgtccat tttctgagaa cctactatgt tttttgtttg agcttaaaaa ttcgattgaa
     3241 caactgtaca gtatagcgtc acttcttagg attcctgata ttttatgctg aactactggt
     3301 tcataaaatt ttctttcaaa taaaactgtc tgatttgttt attacacaac gaagtttgta
     3361 tgcgtaaacg ataaaaacat cttctagaca gaccaaagtc tttcttcaat agtcaaatca
     3421 tttcaaattg ttcctaatga tcctttcatg aatatttgta caaaataaca gttgacaaac
     3481 caaatatttt gtgaaaaatt ggagtgatta gcgaatcccg taggaaaaac caggagtcag
     3541 ttaagtgaag cattttcttg caagaaagaa ataaatgatt tcatgttccc aactggtgat
     3601 aatatctctc taaggagtta tcatgtgttg tttttggaaa ttaatattct tcactttttg
     3661 tgtttttatt tcaaccattc tcgattataa ttttgcttcc agactttttt gtccgaatac
     3721 tcgtgatcca taaaaaccca gctttcttgt acaaagtggt tgatgggacg atatcgggta
     3781 ccggtagaaa aatgtctgaa cttatcaagg aaaacatgca tatgaagctt tacatggaag
     3841 gaaccgttga taaccaccac ttcaagtgca cttctgaagg agagggaaag ccatacgaag
     3901 gaactcaaac catgcgcatc aaggttgttg aaggaggacc acttccattc gctttcgaca
     3961 tccttgccac ctctttcctt tacggatcta aggtaagttt aaacatatat atactaacta
     4021 accctgatta tttaaatttt cagactttca tcaaccatac tcaaggaatt ccagatttct
     4081 tcaagcaatc tttcccagaa ggattcactt gggagcgcgt tactacctac gaggatggag
     4141 gagttcttac cgccacccaa gatacctccc ttcaagatgg atgccttatc tacaacgtta
     4201 agatccgcgg agttaacttc acttctaacg gaccagttat gcaaaagaag actcttggat
     4261 gggaggcttt cactgagacc ctttacccag ccgatggagg acttgaggga cgcaacgaca
     4321 tggctcttaa gcttgttgga ggatcccacc ttatcgctaa catcaaggta agtttaaaca
     4381 gttcggtact aactaaccat acatatttaa attttcagac tacctaccgc tctaagaagc
     4441 cagccaagaa tcttaagatg ccaggagttt attacgtcga ttaccgcctt gagcgcatca
     4501 aggaagctaa caacgagact tacgttgagc aacacgaggt tgctgtcgct cgttactgcg
     4561 atcttccatc taagcttgga cacaagctca actaagctgt ctcatcctac tttcacctag
     4621 ttaactgctt gtcttaaaat ctatgcttct ctttagtatc taaaattttc ctagaagctt
     4681 acaagtatat aaatggtctc ttctcaataa aggttgtata tttattcatc ttattgaatc
     4741 tgccatttcc tcgtttttgc gagtttatat accttccaat tttctttcta ttgtattttc
     4801 aacttctaat tttaattcag ggaaactgct gtaccggtag aaaaaatgga gaccgacacc
     4861 cttcttcttt gggtccttct tctttgggtt ccaggatcta ccggagatgc tccagttgga
     4921 ggaatgtcta agggagaaga gcttttcacc ggagttgttc caatccttgt cgagcttgat
     4981 ggagatgtta acggacacaa gttctctgtt cgtggagagg gagagggaga cgctaccaac
     5041 ggaaagctta ctcttaagtt catctgcact actggaaagc ttccagttcc atggccaacc
     5101 cttgttacca cccttactta cggagttcaa tgcttctctc gttacccaga tcatatgaag
     5161 caacatgatt tcttcaagtc tgctatgcca gagggatacg ttcaagagcg taccatctct
     5221 ttcaaggatg atggaaccta caaggtaagt ttaaacatat atatactaac taaccctgat
     5281 tatttaaatt ttcagacccg tgctgaggtt aagttcgagg gagataccct tgttaaccgt
     5341 atcgagctta agggaatcga tttcaaggaa gatggaaaca tccttggaca taagcttgaa
     5401 tacaatttca ataatccact taacatgaac ttcgagcttt ccgacgagat gaaggctctt
     5461 ttcaaggagc caaacgataa ggctcttaag actggaggat ccggaggatc tggaggatcc
     5521 cgtgccccaa cttattcccc accaccacca ccaggtggtg gatctggagg aggatctgga
     5581 actgaggttc catcttccat gaccaccgag tctaccgcta ccgctatgca atctgagatg
     5641 tctacttcta tcatggagac tactaccacc cttgccacct ctaccgcccg ccgcggaaag
     5701 ccaccaacta aggtaagttt aaacagttcg gtactaacta accatacata tttaaatttt
     5761 caggaaccaa tctctcaaac caccgacgac atcctcgtcg cttccgctga gtgcccatcc
     5821 gatgacgagg atatcgatcc atgcgagcca tcttctggag gacttgctaa cccaacccgc
     5881 gttggaggac gtgagccata cccaggatct gctgaggtta tccgtgagtc ctcttccacc
     5941 accggaatgg ttgttggaat cgtcgctgct gccgcccttt gcatccttat ccttctttac
     6001 gctatgtaca agtaccgcaa ccgcgacgag ggatcctacc acgttgacga gtctcgtaac
     6061 tacatctcta actctgctca atccaacgga gccgttgtta aggagaagca accatcttct
     6121 gctaagtctg ctaataagaa taagaagaac aaggacaagg aatactacgt ctaatctaga
     6181 caattccaac tgagcgccgg tcgctaccat taccaacttg tctggtgtca aaaataatag
     6241 gggccgctgt catcagagta agtttaaact gagttctact aactaacgag taatatttaa
     6301 attttcagca tctcgcgccc gtgcctctga cttctaagtc caattactct tcaacatccc
     6361 tacatgctct ttctccctgt gctcccaccc cctatttttg ttattatcaa aaaacttctc
     6421 ttaatttctt tgttttttag cttcttttaa gtcacctcta acaatgaaat tgtgtagatt
     6481 caaaaataga attaattcgt aataaaaagt cgaaaaaaat tgtgctccct ccccccatta
     6541 ataataattc tatcccaaaa tctacacaat gttctgtgta cacttcttat gttttttact
     6601 tctgataaat ttttttgaaa catcatagaa aaaaccgcac acaaaatacc ttatcatatg
     6661 ttacgtttca gtttatgacc gcaattttta tttcttcgca cgtctgggcc tctcatgacg
     6721 tcaaatcatg ctcatcgtga aaaagttttg gagtattttt ggaatttttc aatcaagtga
     6781 aagtttatga aattaatttt cctgcttttg ctttttgggg tttcccctat tgtttgtcaa
     6841 gatttcgagg acggcgtttt tcttgctaaa atcacaagta ttgatgagca cgatgcaaga
     6901 aagatcggaa gaaggtttgg gtttgaggct cagtggaagg tgagtagaag ttgataattt
     6961 gaaagtggag tagtgtctat ggggtttttg ccttaaatga cagaatacat tcccaatata
     7021 ccaaacataa ctgtttccta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg
     7081 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     7141 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     7201 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     7261 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg
     7321 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     7381 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     7441 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     7501 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     7561 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     7621 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7681 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7741 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7801 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7861 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7921 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7981 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     8041 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     8101 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     8161 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     8221 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     8281 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     8341 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     8401 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     8461 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     8521 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     8581 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     8641 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     8701 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8761 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8821 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     8881 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     8941 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     9001 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     9061 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     9121 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     9181 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     9241 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     9301 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     9361 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     9421 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     9481 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     9541 agctgtaagt ttaaacatga tcttactaac taactattct catttaaatt ttcagagctt
     9601 aaaaatggct gaaatcactc acaacgatgg atacgctaac aacttggaaa tgaaataagc
     9661 tacccat
//
