LOCUS       sequence-446186-        4526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminal His-tagged qmTQ2-glucose, a cyan 
            fluorescent FLIM biosensor for detecting glucose.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4526)
  AUTHORS   Zhong C, Arai S, Okada Y
  TITLE     Development of fluorescence lifetime biosensors for ATP, cAMP, 
            citrate, and glucose using the mTurquoise2-based platform.
  JOURNAL   Cell Rep Methods. 2024 Nov 18;4(11):100902. doi: 
            10.1016/j.crmeth.2024.100902.
  PUBMED    39561716
REFERENCE   2  (bases 1 to 4526)
  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 Rep 
            Methods."; date: "2024-11-18"; volume: "4(11):100902. doi"; pages: "
            10.1016/j.crmeth.2024.100902"
COMMENT     Sequence Label: pRSETA-qmTQ2-glucose
FEATURES             Location/Qualifiers
     source          1..4526
                     /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     complement(262..283)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(523..542)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1615..1866
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1803..1824
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1927..1945)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1941..1988
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2085..2540
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2172..2191)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2382..2403
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2566..2670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2671..2739,2740..3531)
                     /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(2889..2908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3702..4290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4191..4210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4444..4461
                     /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 gggatccgag ctcgagatgg tgagcaaggg cgaggagctg
      241 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      301 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
      361 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gtcctggggc
      421 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      481 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
      541 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
      601 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacggtacc
      661 actgatgggc gcattggtgt aacaatctat aagtacgacg ataactttat gtctgtagtg
      721 cgcaaggcta ttgagcaaga tgcgaaagcc gcgccagatg ttcagctgct gatgaatgat
      781 tctcagaatg accagtccaa gcagaacgat cagatcgacg tattgctggc gaaaggggtg
      841 aaggcactgg caatcaacct ggttgacccg gcagctgcgg gtacggtgat tgagaaagcg
      901 cgtgggcaaa acgtgccggt ggttttcttc aacaaagaac cgtctcgtaa ggcgctggat
      961 agctacgaca aagcctacta cgttggcact gactccaaag agtccggcat tattcaaggc
     1021 gatttgattg ctaaacactg ggcggcgaat cagggttggg atctgaacaa agacggtcag
     1081 attcagttcg tactgctgaa aggtgaaccg ggccatccgg atgcagaagc acgtaccact
     1141 tacgtgatta aagaattgaa cgataaaggc atcaaaactg aacagttaca gttagatacc
     1201 gcaatgtggg acaccgctca ggcgaaagat aagatggacg cctggctgtc tggcccgaac
     1261 gccaacaaaa tcgaagtggt tatcgccaac aacgatgcga tggcaatggg cgcggttgaa
     1321 gcgctgaaag cacacaacaa gtccagcatt ccggtgtttg gcgtcgatgc gctgccagaa
     1381 gcgctggcgc tggtgaaatc cggtgcactg gcgggcaccg tactgaacga tgctaacaac
     1441 caggcgaaag cgacctttga tctggcgaaa aacctggccg atggtaaagg tgcggctgat
     1501 ggcaccaact ggaaaatcga caacaaagtg gtccgcgtac cttatgttgg cgtagataaa
     1561 gacaacctgt atgaggcact ggaattcttc agcgacaacg tctatatcac cgccgacaag
     1621 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     1681 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1741 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     1801 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1861 tacaagtaac gaagcttgat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     1921 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     1981 gttttttgct gaaaggagga actatatccg gatctggcgt aatagcgaag aggcccgcac
     2041 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     2101 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2161 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2221 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2281 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2341 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2401 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2461 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2521 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2581 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2641 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2701 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2761 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2821 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2881 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     2941 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3001 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3061 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3121 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3181 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3241 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3301 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3361 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3421 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3481 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3541 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3601 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3661 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3721 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3781 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3841 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3901 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3961 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4021 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4081 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4141 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4201 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4261 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4321 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4381 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4441 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4501 gcgcgttggc cgattcatta atgcag
//
