LOCUS       sequence-316046-        7059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Red fluorescent protein-based glucose sensor for mammalian cells 
            with lower affinity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7059)
  AUTHORS   Mita M, Sugawara I, Harada K, Ito M, Takizawa M, Ishida K, Ueda H, 
            Kitaguchi T, Tsuboi T
  TITLE     Development of red genetically encoded biosensor for visualization 
            of intracellular glucose dynamics.
  JOURNAL   Cell Chem Biol. 2021 Jun 29. pii: S2451-9456(21)00301-9. doi: 
            10.1016/j.chembiol.2021.06.002.
  PUBMED    34197723
REFERENCE   2  (bases 1 to 7059)
  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: "Cell Chem 
            Biol. 2021 Jun 29. pii: S2451-9456(21)00301-9. doi: 
            10.1016/j.chembiol.2021.06.002."
COMMENT     Sequence Label: Red Glifon 3000
FEATURES             Location/Qualifiers
     source          1..7059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1399..1418
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2653..2670)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2659..2883
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2929..3357
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3016..3035)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3226..3247
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3366..3386)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3371..3700
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3551..3686
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3613..3632
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3767..4561
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3821..3840)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4431..4450
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4735..4856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4772..4791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4826..4845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4905..4921)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4905..4921)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4918..4940)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4929..4945
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4953..4959,4960..4977,4978..4983))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4998..5019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5136..5153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5307..5892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5387..5406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6063..6854,6855..6923))
                     /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     6686..6705
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6924..7028)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcc
      961 accacactgg actagtggat ccatgctggt gagcaagggc ggggaggata acatggccat
     1021 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     1081 cgagatcgag ggcgagggcg agggccgccc ctacgaggcc tttcagaccg ctaagctgaa
     1141 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     1201 cggctccaag gcctacatta agcacccagc cgacatcccc gactacttca agctgtcctt
     1261 ccccgagggc ttcaggtggg agcgcgtgat gaacttcgag gacggcggca ttattcacgt
     1321 taaccaggac tcctccctgc aggacggcgt attcatctac aaggtgaagc tgcgcggcac
     1381 caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggctcttcc
     1441 tggtactgat gggcgcattg gtgtaacaat ctataagtac gacgataact ggatgtctgt
     1501 agtgcgcaag gctattgagc aagatgcgaa agccgcgcca gatgttcagc tgctgatgaa
     1561 tgattctcag aatgaccagt ccaagcagaa cgatcagatc gacgtattgc tggcgaaagg
     1621 ggtgaaggca ctggcaatca acctggttga cccggcagct gcgggtacgg tgattgagaa
     1681 agcgcgtggg caaaacgtgc cggtggtttt cttcaacaaa gaaccgtctc gtaaggcgct
     1741 ggatagctac gacaaagcct actacgttgg cactgactcc aaagagtccg gcattattca
     1801 aggcgatttg attgctaaac actgggcggc gaatcagggt tgggatctga acaaagacgg
     1861 tcagattcag ttcgtactgc tgaaaggtga accgggccat ccggatgcag aagcacgtac
     1921 cacttacgtg attaaagaat tgaacgataa aggcatcaaa actgaacagt tacagttaga
     1981 taccgcaatg tgggacaccg ctcaggcgaa agataagatg gacgcctggc tgtctggccc
     2041 gaacgccaac aaaatcgaag tggttatcgc caacaacgat gcgatggcaa tgggcgcggt
     2101 tgaagcgctg aaagcacaca acaagtccag cattccggtg tttggcgtcg atgcgctgcc
     2161 agaagcgctg gcgctggtga aatccggtgc actggcgggc accgtactga acgatgctaa
     2221 caaccaggcg aaagcgacct ttgatctggc gaaaaacctg gccgatggta aaggtgcggc
     2281 tgatggcacc aactggaaaa tcgacaacaa agtggtccgc gtaccttatg ttggcgtaga
     2341 taaagacaac ctgtatgagg gttgggtttc cgagcggatg taccccgagg acggcgccct
     2401 gaagagcgag atcaagaagg ggctgaggct gaaggacggc ggccactacg ccgccgaggt
     2461 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca tcgtcgacat
     2521 caagttggac atcgtgtccc acaacgagga ctacaccatc gtggaacagt gcgaacgcgc
     2581 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaaaagc ttaagtttaa
     2641 accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2701 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2761 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2821 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     2881 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     2941 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3001 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3061 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3121 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3181 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3241 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3301 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3361 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3421 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3481 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3541 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3601 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3661 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3721 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3781 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3841 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3901 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3961 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4021 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4081 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4141 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4201 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4261 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4321 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4381 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4441 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4501 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4561 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4621 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4681 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4741 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4801 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4861 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4921 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4981 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5041 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5101 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5161 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5221 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5281 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5341 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5401 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5461 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5521 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5581 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5641 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5701 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5761 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5821 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5881 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5941 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6001 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6061 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6121 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6181 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6241 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6301 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6361 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6421 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6481 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6541 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6601 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6661 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6721 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6781 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6841 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6901 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6961 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7021 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
