LOCUS       sequence-321699-        7062 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 7062)
  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 7062)
  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..7062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1165..1184
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2419..2436)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2425..2649
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2695..3123
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2782..2801)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2992..3013
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3132..3152)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3137..3466
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3317..3452
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3379..3398
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3533..4327
                     /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(3587..3606)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4197..4216
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4501..4622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4538..4557)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4592..4611
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4671..4687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4671..4687)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4684..4706)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4695..4711
                     /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(4719..4725,4726..4743,4744..4749))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4764..4785
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4902..4919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5073..5661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5153..5172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5832..6623,6624..6692))
                     /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     6455..6474
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6693..6797)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6872..6891)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacgggccc tctagactcg agcggccgcc actgtgctgg atatctgcag
      721 aattccacca cactggacta gtggatccat gctggtgagc aagggcgggg aggataacat
      781 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
      841 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gaggcctttc agaccgctaa
      901 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
      961 catgtacggc tccaaggcct acattaagca cccagccgac atccccgact acttcaagct
     1021 gtccttcccc gagggcttca ggtgggagcg cgtgatgaac ttcgaggacg gcggcattat
     1081 tcacgttaac caggactcct ccctgcagga cggcgtattc atctacaagg tgaagctgcg
     1141 cggcaccaac ttcccccccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     1201 tcttcctggt actgatgggc gcattggtgt aacaatctat aagtacgacg ataactggat
     1261 gtctgtagtg cgcaaggcta ttgagcaaga tgcgaaagcc gcgccagatg ttcagctgct
     1321 gatgaatgat tctcagaatg accagtccaa gcagaacgat cagatcgacg tattgctggc
     1381 gaaaggggtg aaggcactgg caatcaacct ggttgacccg gcagctgcgg gtacggtgat
     1441 tgagaaagcg cgtgggcaaa acgtgccggt ggttttcttc aacaaagaac cgtctcgtaa
     1501 ggcgctggat agctacgaca aagcctacta cgttggcact gactccaaag agtccggcat
     1561 tattcaaggc gatttgattg ctaaacactg ggcggcgaat cagggttggg atctgaacaa
     1621 agacggtcag attcagttcg tactgctgaa aggtgaaccg ggccatccgg atgcagaagc
     1681 acgtaccact tacgtgatta aagaattgaa cgataaaggc atcaaaactg aacagttaca
     1741 gttagatacc gcaatgtggg acaccgctca ggcgaaagat aagatggacg cctggctgtc
     1801 tggcccgaac gccaacaaaa tcgaagtggt tatcgccaac aacgatgcga tggcaatggg
     1861 cgcggttgaa gcgctgaaag cacacaacaa gtccagcatt ccggtgtttg gcgtcgatgc
     1921 gctgccagaa gcgctggcgc tggtgaaatc cggtgcactg gcgggcaccg tactgaacga
     1981 tgctaacaac caggcgaaag cgacctttga tctggcgaaa aacctggccg atggtaaagg
     2041 tgcggctgat ggcaccaact ggaaaatcga caacaaagtg gtccgcgtac cttatgttgg
     2101 cgtagataaa gacaacctgt atgagggttg ggtttccgag cggatgtacc ccgaggacgg
     2161 cgccctgaag agcgagatca agaaggggct gaggctgaag gacggcggcc actacgccgc
     2221 cgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacatcgt
     2281 cgacatcaag ttggacatcg tgtcccacaa cgaggactac accatcgtgg aacagtgcga
     2341 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aaaagcttaa
     2401 gtttaaaccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2461 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2521 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2581 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2641 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2701 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2761 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2821 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2881 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2941 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3001 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3061 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3121 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3181 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3241 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3301 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3361 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3421 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3481 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3541 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3601 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3661 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3721 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3781 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3841 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3901 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3961 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4021 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4081 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4141 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4201 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4261 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4321 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4381 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4441 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4501 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4561 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4621 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4681 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4741 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4801 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4861 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4921 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4981 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5041 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5101 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5161 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5221 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5281 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5341 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5401 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5461 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5521 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5581 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5641 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5701 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5761 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5821 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5881 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5941 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6001 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6061 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6121 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6181 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6241 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6301 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6361 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6421 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6481 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6541 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6601 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6661 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6721 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6781 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     6841 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     6901 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     6961 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     7021 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt
//
