LOCUS       sequence-327724-        8643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green fluorescent glutamate reporter fused to TARP gamma-8 for 
            mammalian expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8643)
  AUTHORS   Hao Y, Toulme E, Konig B, Rosenmund C, Plested AJR
  TITLE     Targeted sensors for glutamatergic neurotransmission.
  JOURNAL   eLife. 2023 Jan 9;12:e84029. doi: 10.7554/eLife.84029.
  PUBMED    36622100
REFERENCE   2  (bases 1 to 8643)
  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: "eLife."; 
            date: "2023-01-9"; volume: "12:e84029. doi"; pages: " 
            10.7554/eLife.84029"
COMMENT     Sequence Label: SnFR-gamma8
FEATURES             Location/Qualifiers
     source          1..8643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        172..551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        552..755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     706..726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     800..819
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        800..818
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     921..980
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1949..1970
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2076..2097)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2337..2356)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2567..2585
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     complement(4171..4188)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4177..4401
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4447..4875
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4534..4553)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4744..4765
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4884..4904)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4889..5218
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5069..5204
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5131..5150
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5285..6079
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5339..5358)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5949..5968
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6253..6374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6290..6309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6344..6363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6423..6439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6423..6439)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6436..6458)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6447..6463
                     /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)."
     promoter        complement(join(6471..6477,6478..6495,6496..6501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6516..6537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6654..6671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6825..7413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6905..6924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7584..8375,8376..8444))
                     /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     8207..8226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8445..8549)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8624..8643)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
       61 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
      121 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      181 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      241 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      301 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      361 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      421 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      481 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      541 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      601 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      661 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      721 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
      781 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgtt
      841 taaacttaag cttggtaccg aactcggatc cactagtaac ggccgccagt gtgctggaat
      901 tcggcttggg gatatccacc atggagacag acacactcct gctatgggta ctgctgctct
      961 gggttccagg ttccactggt gacagatctg ccgcaggcag cacgctggac aaaatcgcca
     1021 aaaacggtgt gattgtcgtc ggtcaccgtg aatcttcagt gcctttctct tattacgaca
     1081 atcagcaaaa agtggtgggt tactcgcagg attactccaa cgccattgtt gaagcagtga
     1141 aaaagaaact caacaaaccg gacttgcagg taaaactgat tccgattacc tcacaaaacc
     1201 gtattccact gctgcaaaac ggcactttcg attttgaatg tggttctacc accaacaacg
     1261 tcgaacgcca aaaacaggcg gctttctctg acactatttt cgtggtcggt acgcgcctgt
     1321 tgaccaaaaa gggtggcgat atcaaagatt ttgccaacct gaaagacaaa gccgtagtcg
     1381 tcacttccgg cactacctct gaagttttgc tcaacaaact gaatgaagag caaaaaatga
     1441 atatgcgcat catcagcgcc aaagatcacg gtgactcttt ccgcaccctg gaaagcggtc
     1501 gtgccgttgc ctttatgatg gatgacgtgc tgctggccgg tgaacgtgcg aaagcgaaga
     1561 aaccagacaa ctgggaaatc gtcggcaagc cgcagtctca ggaggcctac ggttgtatgt
     1621 tgcgtaaaga tgatccgcag ttcaaaaagc tgatggatga caccatcgct caggtgcaga
     1681 cctccggtga agcggaaaaa tggtttgata agtggttcaa aaatccaatt ctggtgagcc
     1741 acaacgtcta tatcatggcc gacaagcaga ggaacggcat caaggcgaac ttcaagatcc
     1801 gccacaacat cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag aacaccccca
     1861 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccaaacttt
     1921 cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1981 ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc atggtgagca
     2041 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2101 acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac ggcaagctga
     2161 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2221 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2281 tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc ttcaaggacg
     2341 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2401 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcat aagctggagt
     2461 acaactttaa caaccccctg aacatgaatt tcgaactgtc agacgaaatg aaagcactgt
     2521 tcaaagaacc gaatgacaag gcactgaaag gtgggcgcgc ccgggcagat gtgtataaga
     2581 gacaggaaaa gaaaaaagca ggactgtttg aacaaatcac gaagactcat gggacgatca
     2641 ttggtgtcac atcagggatc gtcctggtcc ttctcattat ttctatttta gtacaagtga
     2701 aacagccccg gaaaaaggtc atggcttgca aaactgcatt taataaaact gggttccagg
     2761 aagtatttga tcctcctcat tatgaactat tttcactaag agagaaagag atttctgcag
     2821 acgccgggtc cgagtcattg aaacgctgga atgaagagag ggggttgtgg tgcgaaaagg
     2881 gcgttcaggt actactgacc accataggcg cctttgcagc ttttggcctc atgaccatcg
     2941 ccatcagcac tgactactgg ctctacacaa gagctctcat ctgcaacacc accaacctca
     3001 cagcaggtga tgatggacca ccccatcgtg ggggcagtgg ctcctcagag aagaaggacc
     3061 ctgggggcct cacacattca ggcctctggc ggatatgctg cctggaaggg ttgaaaagag
     3121 gtgtctgcgt gaagatcaac cacttcccgg aggacacgga ctacgaccac gacagcgcgg
     3181 agtacctgct ccgagtagtc cgggcctcca gcatctttcc tatcctgagc gccatcctgc
     3241 tgctgctcgg gggcgtgtgc gtagctgcct ctcgcgtcta caaatccaaa aggaacatca
     3301 tcctgggcgc agggatcctg ttcgtggcag caggcctgag caacatcatc ggggtgattg
     3361 tgtacatatc ggccaacgcc ggcgagcctg gccccaagag ggacgaggag aagaaaaacc
     3421 actactcgta cggctggtcc ttctacttcg gcgggctgtc cttcatcctg gctgaggtga
     3481 tcggagtact ggccgtcaac atctacatcg agcgcagccg cgaggcacac tgccaatcac
     3541 gctcggacct gctcaaggcc ggcggcggcg cgggcggcag tggcgggagc ggcccctcgg
     3601 ccatcctccg tctgcccagt taccgcttcc gctaccgccg ccgctcccgc tccagctccc
     3661 gaggctccag cgaggcgtcg ccatcccggg atgcgtctcc cggcggcccc gggggcccgg
     3721 gcttcgcctc cacggacatc tccatgtaca cgctcagccg cgacccgtcc aagggcagcg
     3781 tggctgcggg gctggcgagc gccggtggcg gcggcagcgg tgccggcgtg ggtgcctacg
     3841 gcggggcggc cggggcggcg gggggcggcg gggcgggctc ggagcgggac cgcgggagct
     3901 cggcgggttt tctcacgctg cacaacgcct tccccaagga agcggcgtcc ggcgtcacgg
     3961 tcacagtcac cggaccgccc gctgcacccg cgcccgcgcc cgcgccgccc gctcctgcag
     4021 cgcccgcgcc cgggaccctg tccaaagagg ccgcggcgtc caacaccaac acgctcaaca
     4081 ggaaaaccac gcccgtgtag gaattctgca gatatccagc acagtggcgg ccgctcgagt
     4141 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     4201 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4261 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4321 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     4381 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     4441 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4501 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4561 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4621 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4681 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4741 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4801 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4861 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4921 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4981 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5041 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5101 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     5161 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     5221 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     5281 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5341 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5401 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5461 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5521 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5581 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5641 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5701 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5761 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5821 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5881 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5941 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6001 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6061 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     6121 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6181 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6241 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6301 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6361 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     6421 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6481 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6541 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6601 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6661 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6721 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6781 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6841 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6901 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6961 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7021 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7081 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7141 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7201 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7261 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7321 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7381 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7441 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7501 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7561 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7621 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7681 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7741 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7801 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7861 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7921 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7981 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     8041 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8101 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8161 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8221 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8281 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8341 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8401 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8461 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8521 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8581 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     8641 ccg
//
