LOCUS       sequence-446764-        4010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminal His-tagged qmTQ2-ATP, a cyan 
            fluorescent FLIM biosensor for ATP imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4010)
  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 4010)
  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-ATP
FEATURES             Location/Qualifiers
     source          1..4010
                     /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             1099..1350
                     /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     1287..1308
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1411..1429)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1425..1472
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1569..2024
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1656..1675)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1866..1887
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2050..2154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2155..2223,2224..3015)
                     /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(2373..2392)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3186..3774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3675..3694
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3928..3945
                     /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 caagcctgtg
      661 atgaagaccg tgaaggtgaa catcaccacc cccgacggcc ccgtgtacga cgccgacatc
      721 gagatggtga gcgtgcgggc cgagagcggc gacctgggca tcctgcccgg ccacatcccc
      781 accaaggccc ccctgaagat cggcgccgtg cggctgaaga aggacggcca gaccgagatg
      841 gtggccgtga gcggcggcaa cgtggaggtg cggcccgacc aggtgaccat caccgcccag
      901 gccgccgaga ccgccgaggg catcgacaag gagcgggccg aggccgcccg gcagcgggcc
      961 caggagcggc tgaacagcca gagcgacgac accgacatcc ggcgggccga gctggccctg
     1021 cagcgggccc tgaaccggct ggacgtggcc gcactgaaga aagaagaatt cttcagcgac
     1081 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1141 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     1201 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc
     1261 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1321 atcactctcg gcatggacga gctgtacaag taacgaagct tgatccggct gctaacaaag
     1381 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     1441 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatctg
     1501 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     1561 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1621 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1681 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1741 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     1801 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1861 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     1921 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     1981 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     2041 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2101 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2161 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2221 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2281 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2341 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2401 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2461 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2521 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2581 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2641 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2701 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2761 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2821 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2881 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2941 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3001 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3061 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3121 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3181 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3241 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3301 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3361 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3421 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3481 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3541 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3601 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3661 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3721 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3781 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3841 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3901 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3961 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
//
