LOCUS       sequence-211677-        6828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fast indicator for extracellular glutamate imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6828)
  AUTHORS   Helassa N, Durst CD, Coates C, Kerruth S, Arif U, Schulze C, Wiegert
            JS, Geeves M, Oertner TG, Torok K
  TITLE     Ultrafast glutamate sensors resolve high-frequency release at 
            Schaffer collateral synapses.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 May 7. pii: 1720648115. doi: 
            10.1073/pnas.1720648115.
  PUBMED    29735711
REFERENCE   2  (bases 1 to 6828)
  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: "Proc Natl 
            Acad Sci U S A. 2018 May 7. pii: 1720648115. doi: 
            10.1073/pnas.1720648115."
COMMENT     Sequence Label: pCMV iGluu
FEATURES             Location/Qualifiers
     source          1..6828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        48..427
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        428..631
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     582..602
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     676..695
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        676..694
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     775..834
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1803..1824
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1930..1951)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2191..2210)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2416..2445
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2449..2595
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(2617..2634)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2623..2847
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2979..3567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3059..3078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3896..3943
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3968..3987
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4175..4969)
                     /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
                     GLAPAELFARLKASKPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4286..4305)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4896..4915
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5068..5087)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        5166..5357
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5314..5334
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5417..6208,6209..6277))
                     /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     6040..6059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6278..6382)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6409..6828,1..36)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6496..6515)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6706..6727
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 acgcgaattt taacaaaata ttaacgctta caatttacgc gcgcgttgac attgattatt
       61 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      121 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      181 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      241 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      301 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      361 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      421 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      481 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      541 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      601 tgtacggtgg gaggtctata taagcagaac tctctggcta actagagaac ccactgctta
      661 ctggcttatc gaaattaata cgactcacta tagggagacc caagcttggt accgaactcg
      721 gatccactag taacggccgc cagtgtgctg gaattcggct tggggatatc caccatggag
      781 acagacacac tcctgctatg ggtactgctg ctctgggttc caggttccac tggtgacaga
      841 tctgccgcag gcagcacgct ggacaaaatc gccaaaaacg gtgtgattgt cgtcggtcac
      901 cgtgaatctt cagtgccttt ctcttattac gacaatcagc aaaaagtggt gggttactcg
      961 caggattact ccaacgccat tgttgaagca gtgaaaaaga aactcaacaa accggacttg
     1021 caggtaaaac tgattccgat taccacacaa aaccgtattc cactgctgca aaacggcact
     1081 ttcgattttg aatgtggttc taccaccaac aacgtcgaac gccaaaaaca ggcggctttc
     1141 tctgacacta ttttcgtggt cggtacgcgc ctgttgacca aaaagggtgg cgatatcaaa
     1201 gattttgcca acctgaaaga caaagccgta gtcgtcactt ccggcactac ctctgaagtt
     1261 ttgctcaaca aactgaatga agagcaaaaa atgaatatgc gcatcatcag cgccaaagat
     1321 cacggtgact ctttccgcac cctggaaagc ggtcgtgccg ttgcctttat gatggatgac
     1381 gtgctgctgg ccggtgaacg tgcgaaagcg aagaaaccag acaactggga aatcgtcggc
     1441 aagccgcagt ctcaggaggc ctacggttgt atgttgcgta aagatgatcc gcagttcaaa
     1501 aagctgatgg atgacaccat cgctcaggtg cagacctccg gtgaagcgga aaaatggttt
     1561 gataagtggt tcaaaaatcc aattctggtg agccacaacg tctatatcat ggccgacaag
     1621 cagaggaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcggcgtg
     1681 cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1741 gacaaccact acctgagcac ccagtccaaa ctttcgaaag accccaacga gaagcgcgat
     1801 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1861 tacaagggcg gtaccggagg gagcatggtg agcaagggcg aggagctgtt caccggggtg
     1921 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1981 gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2041 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2101 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2161 tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2221 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2281 gaggacggca acatcctggg gcataagctg gagtacaact ttaacaaccc cctgaacatg
     2341 aatttcgaac tgtcagacga aatgaaagca ctgttcaaag aaccgaatga caaggcactg
     2401 aaactgcagg tcgacgaaca aaaactcatc tcagaagagg atctgaatgc tgtgggccag
     2461 gacacgcagg aggtcatcgt ggtgccacac tccttgccct ttaaggtggt ggtgatctca
     2521 gccatcctgg ccctggtggt gctcaccatc atctccctta tcatcctcat catgctttgg
     2581 cagaagaagc cacgttaggc ggccgctcga gatcagcctc gactgtgcct tctagttgcc
     2641 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2701 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2761 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2821 gtgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagt ggcggtaata
     2881 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2941 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3001 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3061 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3121 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3181 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3241 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3301 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3361 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3421 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3481 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3541 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3601 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3661 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3721 taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc tgggccttca
     3781 cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg gccccaatgg
     3841 ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg tttatgaaca
     3901 aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc gcgggttcct
     3961 tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc gaagaactcc
     4021 agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag
     4081 cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc
     4141 gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     4201 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     4261 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     4321 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     4381 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcttgc
     4441 tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     4501 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     4561 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     4621 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     4681 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     4741 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc agttcattca
     4801 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     4861 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     4921 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     4981 ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa gcctcctcac
     5041 tacttctgga atagctcaga ggccgggcct cggcctctgc ataaataaaa aaaattagtc
     5101 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     5161 ggggcgggac tatggttgct gactaattga gatgcttgct ttgcatactt ctgcctgctg
     5221 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcttg ctttgcatac
     5281 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct
     5341 ggttctttcc gcctcaggac tcttcctttt tcaataaatc aatctaaagt atatatgagt
     5401 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5461 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5521 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5581 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5641 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5701 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5761 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5821 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5881 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5941 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6001 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6061 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6121 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6181 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6241 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6301 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6361 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     6421 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     6481 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     6541 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6601 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     6661 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6721 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6781 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaattta
//
