LOCUS       sequence-327723-        8343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green fluorescent glutamate reporter fused to TARP gamma-2 for 
            mammalian expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8343)
  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 8343)
  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-gamma2
FEATURES             Location/Qualifiers
     source          1..8343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     750..809
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1778..1799
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1905..1926)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2166..2185)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2396..2414
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     complement(3700..3717)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3706..3930
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3976..4404
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4063..4082)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4273..4294
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4413..4433)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4418..4747
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4598..4733
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4660..4679
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4814..5608
                     /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(4868..4887)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5478..5497
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5782..5903
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5819..5838)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5873..5892
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5952..5968)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5952..5968)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5965..5987)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5976..5992
                     /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(6000..6006,6007..6024,6025..6030))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6045..6066
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6183..6200)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6354..6942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6434..6453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7113..7904,7905..7973))
                     /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     7736..7755
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7974..8078)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8153..8172)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga actcggatcc actagtaacg gccgccagtg
      721 tgctggaatt cggcttgggg atatccacca tggagacaga cacactcctg ctatgggtac
      781 tgctgctctg ggttccaggt tccactggtg acagatctgc cgcaggcagc acgctggaca
      841 aaatcgccaa aaacggtgtg attgtcgtcg gtcaccgtga atcttcagtg cctttctctt
      901 attacgacaa tcagcaaaaa gtggtgggtt actcgcagga ttactccaac gccattgttg
      961 aagcagtgaa aaagaaactc aacaaaccgg acttgcaggt aaaactgatt ccgattacct
     1021 cacaaaaccg tattccactg ctgcaaaacg gcactttcga ttttgaatgt ggttctacca
     1081 ccaacaacgt cgaacgccaa aaacaggcgg ctttctctga cactattttc gtggtcggta
     1141 cgcgcctgtt gaccaaaaag ggtggcgata tcaaagattt tgccaacctg aaagacaaag
     1201 ccgtagtcgt cacttccggc actacctctg aagttttgct caacaaactg aatgaagagc
     1261 aaaaaatgaa tatgcgcatc atcagcgcca aagatcacgg tgactctttc cgcaccctgg
     1321 aaagcggtcg tgccgttgcc tttatgatgg atgacgtgct gctggccggt gaacgtgcga
     1381 aagcgaagaa accagacaac tgggaaatcg tcggcaagcc gcagtctcag gaggcctacg
     1441 gttgtatgtt gcgtaaagat gatccgcagt tcaaaaagct gatggatgac accatcgctc
     1501 aggtgcagac ctccggtgaa gcggaaaaat ggtttgataa gtggttcaaa aatccaattc
     1561 tggtgagcca caacgtctat atcatggccg acaagcagag gaacggcatc aaggcgaact
     1621 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgcctaccac taccagcaga
     1681 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     1741 ccaaactttc gaaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1801 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggtacc ggagggagca
     1861 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1921 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg
     1981 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2041 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2101 agcacgactt cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct
     2161 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2221 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcata
     2281 agctggagta caactttaac aaccccctga acatgaattt cgaactgtca gacgaaatga
     2341 aagcactgtt caaagaaccg aatgacaagg cactgaaagg tgggcgcgcc cgggcagatg
     2401 tgtataagag acaggaaaag aaaaaagcag gactgtttga acaaatcacg aagactcatg
     2461 ggacgatcat tggtgtcaca tcagggatcg tcctggtcct tctcattatt tctattttag
     2521 tacaagtgaa acagccccgg aaaaaggtca tggcttgcaa aactgcattt aataaaactg
     2581 ggttccagga agtatttgat cctcctcatt atgaactatt ttcactaaga gagaaagaga
     2641 tttctgcaga cgccgggtcc gggctgtttg atcgaggtgt tcaaatgctt ttaaccaccg
     2701 ttggtgcttt cgctgccttc agcttgatga ccatcgctgt gggaaccgac tattggctgt
     2761 actccagagg ggtttgcaag accaaaagtg tcagtgagaa tgaaaccagc aagaagaacg
     2821 aggaagttat gacccattcc ggattatgga gaacctgctg cctcgaaggg aacttcaaag
     2881 gtctgtgcaa gcaaatcgac cactttccgg aagacgcgga ctacgaagct gacaccgcag
     2941 agtatttcct ccgggccgtg agggcctcga gtatcttccc gatcctgagt gtgatcctgc
     3001 ttttcatggg tggcctctgc atcgcggcga gcgagttcta caagacacgc cacaacatca
     3061 tcctgagtgc tggcatcttc ttcgtgtctg caggtcttag taatatcatc gggatcatcg
     3121 tgtatatatc agccaatgct ggagacccct ccaagagtga ctccaaaaag aacagctact
     3181 cctacggctg gtccttctac ttcggggccc tgtccttcat catcgccgag atggtcgggg
     3241 tgctggccgt gcacatgttt atcgaccgcc acaaacagct gcgggccacg gcccgcgcca
     3301 ccgactacct ccaggcctcc gccatcaccc gcatccccag ctaccgctac cgctaccagc
     3361 gccgcagccg ctccagctcg cgctccaccg agccctctca ctccagagac gcctcgcccg
     3421 tgggcgtgaa gggcttcaac accctgccgt ccacggagat ctccatgtac accctcagta
     3481 gggaccccct gaaggctgcc accacgccca ccgccaccta caactcggac agggataaca
     3541 gctttctcca ggtccacaac tgtatccaga aggacagcaa ggactctctc cacgccaaca
     3601 cagccaaccg ccggaccacg cccgtatgag aattctgcag atatccagca cagtggcggc
     3661 cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     3721 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3781 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3841 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3901 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3961 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4021 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4081 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4141 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4201 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4261 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4321 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4381 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     4441 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4501 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4561 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4621 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4681 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4741 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     4801 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4861 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4921 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4981 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5041 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5101 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5161 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5221 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5281 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5341 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5401 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5461 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5521 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5581 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5641 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5701 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5761 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5821 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5881 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5941 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6001 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6061 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6121 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6181 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6241 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6301 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6361 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6421 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6481 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6541 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6601 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6661 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6721 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6781 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6841 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6901 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6961 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7021 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7081 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7141 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7201 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7261 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7321 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7381 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7441 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7501 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7561 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7621 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7681 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7741 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7801 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7861 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7921 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7981 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8041 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8101 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     8161 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     8221 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     8281 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     8341 gtt
//
