LOCUS       sequence-209538-        6909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Red fluorescent glutamate sensor with ncp topology. 
            Membrane-displayed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6909)
  AUTHORS   Wu J, Abdelfattah AS, Zhou H, Ruangkittisakul A, Qian Y, Ballanyi K,
            Campbell RE
  TITLE     Genetically Encoded Glutamate Indicators with Altered Color and 
            Topology.
  JOURNAL   ACS Chem Biol. 2018 Jan 22. doi: 10.1021/acschembio.7b01085.
  PUBMED    29308878
REFERENCE   2  (bases 1 to 6909)
  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: "ACS Chem 
            Biol. 2018 Jan 22. doi: 10.1021/acschembio.7b01085."
COMMENT     Sequence Label: pDisplay-Rncp-iGluSnFR1
FEATURES             Location/Qualifiers
     source          1..6909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        468..847
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        848..1051
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1002..1022
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1096..1115
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1096..1114
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     1195..1254
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1674..1693
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1798..1815
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1819..1851
                     /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(1855..1863,1864..1878)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             2917..2946
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2950..3096
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(3118..3135)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3124..3348
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3480..4068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3560..3579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4397..4444
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4469..4488
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4676..5470)
                     /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(4787..4806)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5397..5416
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5569..5588)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        5667..5858
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5815..5835
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5918..6709,6710..6778))
                     /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     6541..6560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6779..6883)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttacgc gcgcgttgac attgattatt
      481 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      541 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      601 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      661 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      721 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      781 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      841 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      901 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      961 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     1021 tgtacggtgg gaggtctata taagcagaac tctctggcta actagagaac ccactgctta
     1081 ctggcttatc gaaattaata cgactcacta tagggagacc caagcttggt accgaactcg
     1141 gatccactag taacggccgc cagtgtgctg gaattcggct tggggatatc caccatggag
     1201 acagacacac tcctgctatg ggtactgctg ctctgggttc caggttccac tggtgacaga
     1261 tctgtgagca agggcgaggg ggataacaag gccatcatca aggagttcat gcgcttcaag
     1321 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     1381 cgcccctacg aggcctttca gaccgctaag ctgaaggtga ccaagggtgg ccccctgccc
     1441 ttcgcctggg acatcctgtc tcctcagttc atgtacggct ccaaggccta cattaagcac
     1501 ccagccgaca tccccgacta cttcaagctg tccttccccg agggcttcag gtgggagcgc
     1561 gtgatgatct tcgaggacgg cggcattatt cacgttagcc aggactcctc cctgcaggac
     1621 ggcgtattca tctacaaggt gaagctgcgc ggcaccaact tcccccccga cggccccgta
     1681 atgcagaaga agaccatggg ctgggagcct acgcgtctga acatgaattt cgaactgtca
     1741 gacgaaatga aagcacggtt caaagaaccg aatgacaagg cactgaaagg cggctctcat
     1801 catcatcatc atcatggtat ggctagcatg actggtggac agcaaatggg tcgggatctg
     1861 tacgacgatg acgataagga tcctggctcg agcggatccg ccgcaggcag tacgctggac
     1921 aaaatcgcca aaaacggtgt gattgtcgtc ggtcaccgtg aatcttcagt gcctttctct
     1981 ttttacgaca atcagcaaaa agtggtgggt tactcgcagg attactccaa cgccattgtt
     2041 gaagcagtga agaagaaact caacaaaccg gacttgcagg taaaactgat tccgattacc
     2101 tcacaaaacc gtattccact gctgcaaaac ggcactatcg attttgaatg tggttctacc
     2161 accaacaacg tcgaacgcca aaaacaggcg gctttctctg acactatttt cgtggtcggt
     2221 acgcgcctgt tgaccaaaaa gggtggcgat atcaaagatt ttgccaacct gaaagacaaa
     2281 gccgtagtcg tcacttccgg cactacctct gaagttttgc tcaacaaact gaatgaagag
     2341 caaaaaatga atatgcgcat catcagcgcc aaagatcacg gtgactcttt ccgcaccctg
     2401 gaaagcggtc gtgccgttgc ctttatgatg gatgacgtgc tgctggccgg tgaacgtgcg
     2461 aaagcgaaga aaccagacaa ctgggaaatc gtcggcaagc cgcagtctct ggaggcctac
     2521 ggttgtatgt tgcgtaaaga tgatccgcag ttcaaaaagc tggtggatga caccatcgct
     2581 caggtgcaga cctccggtga agcggaaaaa tggtttgata agtggttcaa aaatccaatt
     2641 gaggtggttt ccgagcggat gtaccccgag gacggcgccc tgaagagcgt gatcaggaag
     2701 gggctgaggc tgaaggacgg cggccactac gccgccgagg ccaagaccac ctacaaggcc
     2761 aagaagcccg tgcagctgcc cggcgcctac atcgtggata tcaagttgga catcgtgtcc
     2821 cacaacgagg actacaccat cgtggaacag tgcgaacgcg ccgagggccg ccactccacc
     2881 ggcggcatgg acgagctgta caagctgcag gtcgacgaac aaaaactcat ctcagaagag
     2941 gatctgaatg ctgtgggcca ggacacgcag gaggtcatcg tggtgccaca ctccttgccc
     3001 tttaaggtgg tggtgatctc agccatcctg gccctggtgg tgctcaccat catctccctt
     3061 atcatcctca tcatgctttg gcagaagaag ccacgttagg cggccgctcg agatcagcct
     3121 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3181 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3241 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3301 attgggaaga caatagcagg cgtgctgggg atgcggtggg ctctatggct tctgaggcgg
     3361 aaagaaccag tggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3421 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3481 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3541 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3601 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3661 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3721 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3781 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3841 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3901 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3961 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4021 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4081 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4141 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4201 tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg
     4261 gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg
     4321 cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac
     4381 cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt
     4441 gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc
     4501 ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
     4561 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
     4621 cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga
     4681 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     4741 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     4801 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     4861 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     4921 gatcctcgcc gtcgggcttg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     4981 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     5041 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     5101 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     5161 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     5221 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     5281 cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     5341 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     5401 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     5461 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg
     5521 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgggcc tcggcctctg
     5581 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     5641 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcttgc
     5701 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     5761 tgagatgctt gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     5821 actgacacac attccacagc tggttctttc cgcctcagga ctcttccttt ttcaataaat
     5881 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5941 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6001 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6061 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6121 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6181 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6241 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6301 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6361 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6421 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6481 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6541 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6601 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6661 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6721 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6781 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6841 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6901 tgccacctg
//
