LOCUS       sequence-260986-        4001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mutant of QUEEN-7+D20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4001)
  AUTHORS   Yaginuma H,  Okada Y
  TITLE     Live cell imaging of metabolic heterogeneity by quantitative 
            fluorescent ATP indicator protein, QUEEN-37C
  JOURNAL   bioRxiv 2021.10.08.463131
REFERENCE   2  (bases 1 to 4001)
  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: "bioRxiv 
            2021.10.08.463131"
COMMENT     Sequence Label: pRSET-QUE7mu R122A
FEATURES             Location/Qualifiers
     source          1..4001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             93..110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             114..146
                     /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(150..158,159..173)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             537..788
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     725..746
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(852..873)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1113..1132)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1383..1401)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1397..1444
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1541..1996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1628..1647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1838..1859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2022..2126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2127..2195,2196..2987)
                     /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(2345..2364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3158..3746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3647..3666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3900..3917
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat ggtatggcta
      121 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatccga
      181 tgaaaacgat ccacgtgagc gtcactactc ctgatggccc ggtgtacgaa gacgatgttg
      241 aaatggtcag cgtcaaagcg aaaagcggcg agctcggcat tttgccgggg cacattccgc
      301 ttaaggcccc gctcgagatc agcgcggccc ggctgaaaaa aggcggcaaa acgcaataca
      361 ttgccgtcag cggcggcaat ttggaagtcc gcccggacaa agtgacgatt aacgctcaag
      421 ctgctgaacg ggcggaggac attgacgtcc tccgcgccaa agcggcgaaa gagcgggcgg
      481 agcgccgcct gcaaagccag acccgttaca acagccacaa cgtctatatc atggccgaca
      541 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
      601 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
      661 ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg
      721 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
      781 tgtacaaggg tggcaccgga gggagcatgg tgagcaaggg cgaggagctg ttcaccgggg
      841 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      901 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      961 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     1021 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1081 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1141 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1201 aggaggacgg caacatcctg gggcacaagc tggagtacaa cctgggtgac atcgacttca
     1261 aacgggccga actggcgtta aaagctgcca tgaaccgttt gagcgttgcg gaaatgaagt
     1321 gaattcgaag cttgatccgg ttgctaacaa agcccgaaag gaagctgagt tggctgctgc
     1381 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1441 tttgctgaaa ggaggaacta tatccggatc tggcgtaata gcgaagaggc ccgcaccgat
     1501 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     1561 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1621 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1681 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1741 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     1801 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     1861 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1921 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     1981 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2041 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2101 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2161 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2221 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2281 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2341 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2401 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2461 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2521 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2581 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2641 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     2701 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2761 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2821 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2881 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2941 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3001 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3061 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3121 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3181 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3241 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3301 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3361 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3421 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3481 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3541 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3601 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3661 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3721 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3781 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3841 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3901 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     3961 gttggccgat tcattaatgc aggatctcga tcccgcgaaa t
//
