LOCUS       sequence-56305-e        4432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a single-wavelength intracellular glutamate sensor constructed from 
            E. coli Gltl and cpGFP. For bacterial cytoplasmic expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4432)
  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 4432)
  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"; doi: 
            "10.1038/nmeth.2333"; journalName: "Nat Methods"; date: "2013-02"; 
            volume: "10"; issue: "2"; pages: "162-70"
COMMENT     Sequence Label: pRSET.GltI253-cpGFP.L1LV/L2NP
FEATURES             Location/Qualifiers
     source          1..4432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(169..177,178..192)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     1167..1188
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1294..1315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1555..1574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1833..1851)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1847..1894
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1991..2446
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2078..2097)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2288..2309
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2472..2576
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2577..2645,2646..3437)
                     /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(2795..2814)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3608..4196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4097..4116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4350..4367
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatcgatg gggatccgcc gcaggcagca cgctggacaa aatcgccaaa
      241 aacggtgtga ttgtcgtcgg tcaccgtgaa tcttcagtgc ctttctctta ttacgacaat
      301 cagcaaaaag tggtgggtta ctcgcaggat tactccaacg ccattgttga agcagtgaaa
      361 aagaaactca acaaaccgga cttgcaggta aaactgattc cgattacctc acaaaaccgt
      421 attccactgc tgcaaaacgg cactttcgat tttgaatgtg gttctaccac caacaacgtc
      481 gaacgccaaa aacaggcggc tttctctgac actattttcg tggtcggtac gcgcctgttg
      541 accaaaaagg gtggcgatat caaagatttt gccaacctga aagacaaagc cgtagtcgtc
      601 acttccggca ctacctctga agttttgctc aacaaactga atgaagagca aaaaatgaat
      661 atgcgcatca tcagcgccaa agatcacggt gactctttcc gcaccctgga aagcggtcgt
      721 gccgttgcct ttatgatgga tgacgtgctg ctggccggtg aacgtgcgaa agcgaagaaa
      781 ccagacaact gggaaatcgt cggcaagccg cagtctcagg aggcctacgg ttgtatgttg
      841 cgtaaagatg atccgcagtt caaaaagctg atggatgaca ccatcgctca ggtgcagacc
      901 tccggtgaag cggaaaaatg gtttgataag tggttcaaaa atccaattct ggtgagccac
      961 aacgtctata tcatggccga caagcagagg aacggcatca aggcgaactt caagatccgc
     1021 cacaacatcg aggacggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc
     1081 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caaactttcg
     1141 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1201 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat ggtgagcaag
     1261 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1321 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc
     1381 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1441 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1501 ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac
     1561 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1621 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcataa gctggagtac
     1681 aactttaaca accccctgaa catgaatttc gaactgtcag acgaaatgaa agcactgttc
     1741 aaagaaccga atgacaaggc actgaaataa gaattcgaag cttgatccgg ctgctaacaa
     1801 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     1861 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatc
     1921 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     1981 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2041 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2101 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2161 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2221 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2281 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2341 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2401 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     2461 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2521 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2581 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2641 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2701 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2761 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2821 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2881 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2941 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3001 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3061 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3121 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3181 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3241 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3301 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3361 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3421 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3481 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3541 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3601 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3661 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3721 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3781 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3841 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3901 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3961 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4021 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4081 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4141 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4201 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4261 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4321 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4381 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc ag
//
