LOCUS       sequence-209539-        6918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Noncircularly permutated (ncp) variant of iGluSnFR. 
            Membrane-displayed..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6918)
  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 6918)
  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-Gncp-iGluSnFR
FEATURES             Location/Qualifiers
     source          1..6918
                     /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     complement(1306..1327)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1567..1586)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1789..1806
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1810..1842
                     /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(1846..1854,1855..1869)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     2850..2871
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2926..2955
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2959..3105
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(3127..3144)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3133..3357
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3489..4077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3569..3588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4406..4453
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4478..4497
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4685..5479)
                     /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(4796..4815)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5406..5425
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5578..5597)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        5676..5867
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5824..5844
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5927..6718,6719..6787))
                     /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     6550..6569
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6788..6892)
                     /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 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1321 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac
     1381 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1441 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1501 cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc
     1561 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1621 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcat
     1681 aagctggagt acaactttaa caaccccctg aacatgaatt tcgaactgtc agacgaaatg
     1741 aaagcactgt tcaaagaacc gaatgacaag gcactgaaag gcggctctca tcatcatcat
     1801 catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat
     1861 gacgataagg atcctggctc gagcggatcc gccgcaggca gcacgctgga caaaatcgcc
     1921 aaaaacggtg tgattgtcgt cggtcaccgt gaatcttcag tgcctttctc ttattacgac
     1981 aatcagcaaa aagtggtggg ttactcgcag gattactcca acgccattgt tgaagcagtg
     2041 aaaaagaaac tcaacaaacc ggacttgcag gtaaaactga ttccgattac ctcacaaaac
     2101 cgtattccac tgctgcaaaa cggcactttc gattttgaat gtggttctac caccaacaac
     2161 gtcgaacgcc aaaaacaggc ggctttctct gacactattt tcgtggtcgg tacgcgcctg
     2221 ttgaccaaaa agggtggcga tatcaaagat tttgccaacc tgaaagacaa agccgtagtc
     2281 gtcacttccg gcactacctc tgaagttttg ctcaacaaac tgaatgaaga gcaaaaaatg
     2341 aatatgcgca tcatcagcgc caaagatcac ggtgactctt tccgcaccct ggaaagcggt
     2401 cgtgccgttg cctttatgat ggatgacgtg ctgctggccg gtgaacgtgc gaaagcgaag
     2461 aaaccagaca actgggaaat cgtcggcaag ccgcagtctc aggaggccta cggttgtatg
     2521 ttgcgtaaag atgatccgca gttcaaaaag ctgatggatg acaccatcgc tcaggtgcag
     2581 acctccggtg aagcggaaaa atggtttgat aagtggttca aaaatccaat tctggtgagc
     2641 cacaacgtct atatcatggc cgacaagcag aggaacggca tcaaggcgaa cttcaagatc
     2701 cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc
     2761 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaaactt
     2821 tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2881 gggatcactc tcggcatgga cgagctgtac aagctgcagg tcgacgaaca aaaactcatc
     2941 tcagaagagg atctgaatgc tgtgggccag gacacgcagg aggtcatcgt ggtgccacac
     3001 tccttgccct ttaaggtggt ggtgatctca gccatcctgg ccctggtggt gctcaccatc
     3061 atctccctta tcatcctcat catgctttgg cagaagaagc cacgttaggc ggccgctcga
     3121 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3181 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3241 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3301 agggggagga ttgggaagac aatagcaggc gtgctgggga tgcggtgggc tctatggctt
     3361 ctgaggcgga aagaaccagt ggcggtaata cggttatcca cagaatcagg ggataacgca
     3421 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3481 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3541 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3601 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3661 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3721 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3781 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3841 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3901 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3961 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4021 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4081 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4141 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4201 agttttaaat caatctaaag tatatatgag taacctgagg ctatggcagg gcctgccgcc
     4261 ccgacgttgg ctgcgagccc tgggccttca cccgaacttg gggggtgggg tggggaaaag
     4321 gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg gggtatcgac agagtgccag
     4381 ccctgggacc gaaccccgcg tttatgaaca aacgacccaa caccgtgcgt tttattctgt
     4441 ctttttattg ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt
     4501 agcctccccc tagggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc
     4561 cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa
     4621 tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc
     4681 ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg
     4741 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
     4801 cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga
     4861 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg
     4921 tcacgacgag atcctcgccg tcgggcttgc tcgccttgag cctggcgaac agttcggctg
     4981 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
     5041 gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
     5101 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
     5161 ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg
     5221 cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata
     5281 gccgcgctgc ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa
     5341 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
     5401 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
     5461 atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa
     5521 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgggcct
     5581 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     5641 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     5701 gatgcttgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     5761 ctgactaatt gagatgcttg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     5821 cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcaggac tcttcctttt
     5881 tcaataaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5941 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6001 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6061 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6121 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6181 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6241 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6301 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6361 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6421 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6481 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6541 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6601 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6661 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6721 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6781 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6841 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6901 cccgaaaagt gccacctg
//
