LOCUS       40924_11695        6828 bp DNA     circular SYN 13-MAY-2021
DEFINITION  a single-wavelength extracellular glutamate sensor constructed from 
            E. coli Gltl and cpGFP. Membrane-displayed..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6828)
  AUTHORS   Marvin JS, Borghuis BG, Tian L, Cichon J, Harnett MT, Akerboom J, 
            Gordus A, Renninger SL, Chen TW, Bargmann CI, Orger MB, Schreiter 
            ER, Demb JB, Gan WB, Hires SA, Looger LL
  TITLE     An optimized fluorescent probe for visualizing glutamate 
            neurotransmission.
  JOURNAL   Nat Methods. 2013 Feb;10(2):162-70. doi: 10.1038/nmeth.2333. Epub 
            2013 Jan 13.
  PUBMED    23314171
REFERENCE   2  (bases 1 to 6828)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6828)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nmeth.2333"; journalName: "Nat Methods"; date: "2013-02"; 
            volume: "10"; issue: "2"; pages: "162-70"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        637..655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     736..798
                     /product="leader sequence from mouse immunoglobulin kappa
                     light chain"
                     /label=Ig-kappa leader
     CDS             1576..1827
                     /codon_start=1
                     /label=VC155
                     /note="C-terminal fragment of mVenus for use in bimolecular
                     fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
                     /translation="DKQRNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNH
                     YLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     CDS             1846..2289
                     /codon_start=1
                     /label=VN155(I152L)
                     /note="improved N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNN"
     CDS             2377..2406
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2410..2556
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     polyA_signal    2584..2808
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2940..3528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(3857..3904)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3929..3948
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4139..4930)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASKPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4965..5318)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             complement(5381..6238)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6239..6343)
                     /label=AmpR promoter
     rep_origin      6370..6825
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cgcgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagaa ctctctggct
      601 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      661 ccaagcttgg taccgaactc ggatccacta gtaacggccg ccagtgtgct ggaattcggc
      721 ttggggatat ccaccatgga gacagacaca ctcctgctat gggtactgct gctctgggtt
      781 ccaggttcca ctggtgacag atctgccgca ggcagcacgc tggacaaaat cgccaaaaac
      841 ggtgtgattg tcgtcggtca ccgtgaatct tcagtgcctt tctcttatta cgacaatcag
      901 caaaaagtgg tgggttactc gcaggattac tccaacgcca ttgttgaagc agtgaaaaag
      961 aaactcaaca aaccggactt gcaggtaaaa ctgattccga ttacctcaca aaaccgtatt
     1021 ccactgctgc aaaacggcac tttcgatttt gaatgtggtt ctaccaccaa caacgtcgaa
     1081 cgccaaaaac aggcggcttt ctctgacact attttcgtgg tcggtacgcg cctgttgacc
     1141 aaaaagggtg gcgatatcaa agattttgcc aacctgaaag acaaagccgt agtcgtcact
     1201 tccggcacta cctctgaagt tttgctcaac aaactgaatg aagagcaaaa aatgaatatg
     1261 cgcatcatca gcgccaaaga tcacggtgac tctttccgca ccctggaaag cggtcgtgcc
     1321 gttgccttta tgatggatga cgtgctgctg gccggtgaac gtgcgaaagc gaagaaacca
     1381 gacaactggg aaatcgtcgg caagccgcag tctcaggagg cctacggttg tatgttgcgt
     1441 aaagatgatc cgcagttcaa aaagctgatg gatgacacca tcgctcaggt gcagacctcc
     1501 ggtgaagcgg aaaaatggtt tgataagtgg ttcaaaaatc caattctggt gagccacaac
     1561 gtctatatca tggccgacaa gcagaggaac ggcatcaagg cgaacttcaa gatccgccac
     1621 aacatcgagg acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc
     1681 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa actttcgaaa
     1741 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1801 actctcggca tggacgagct gtacaagggc ggtaccggag ggagcatggt gagcaagggc
     1861 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1921 cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca cctacggcaa gctgaccctg
     1981 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2041 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2101 aagtccgcca tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc
     2161 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2221 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcataagct ggagtacaac
     2281 tttaacaacc ccctgaacat gaatttcgaa ctgtcagacg aaatgaaagc actgttcaaa
     2341 gaaccgaatg acaaggcact gaaactgcag gtcgacgaac aaaaactcat ctcagaagag
     2401 gatctgaatg ctgtgggcca ggacacgcag gaggtcatcg tggtgccaca ctccttgccc
     2461 tttaaggtgg tggtgatctc agccatcctg gccctggtgg tgctcaccat catctccctt
     2521 atcatcctca tcatgctttg gcagaagaag ccacgttagg cggccgctcg agatcagcct
     2581 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2641 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2701 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2761 attgggaaga caatagcagg cgtgctgggg atgcggtggg ctctatggct tctgaggcgg
     2821 aaagaaccag tggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2881 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2941 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3001 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3061 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3121 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3181 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3241 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3301 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3361 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3421 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3481 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3541 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3601 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3661 tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg
     3721 gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg
     3781 cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac
     3841 cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt
     3901 gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc
     3961 ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
     4021 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
     4081 cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga
     4141 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     4201 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     4261 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     4321 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     4381 gatcctcgcc gtcgggcttg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     4441 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     4501 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     4561 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     4621 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     4681 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     4741 cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     4801 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     4861 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     4921 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg
     4981 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgggcc tcggcctctg
     5041 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     5101 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcttgc
     5161 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     5221 tgagatgctt gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     5281 actgacacac attccacagc tggttctttc cgcctcagga ctcttccttt ttcaataaat
     5341 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5401 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5461 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5521 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5581 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5641 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5701 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5761 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5821 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5881 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5941 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6001 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6061 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6121 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6181 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6241 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6301 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6361 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6421 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     6481 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6541 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     6601 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6661 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     6721 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     6781 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttacg
//
