LOCUS       sequence-259972-        4000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mutant of QUEEN-7mu.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4000)
  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 4000)
  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 QUEEN7mu K94A
FEATURES             Location/Qualifiers
     source          1..4000
                     /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..1995
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     1837..1858
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2021..2125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2126..2194,2195..2986)
                     /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(2344..2363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3157..3745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3646..3665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3899..3916
                     /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 tccgcgccgc ggcggcgaaa 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 aaacgcgcca tgaaccgttt gagcgttgcg gaaatgaagt
     1321 gaattcgaag cttgatccgg ctgctaacaa 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 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1681 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     1741 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     1801 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     1861 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     1921 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     1981 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2041 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2101 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2161 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2221 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2281 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2341 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2401 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2461 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2521 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2581 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2641 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2701 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2761 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2821 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2881 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2941 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3001 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3061 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3121 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3181 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3241 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3301 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3361 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3421 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3481 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3541 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3601 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3661 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3721 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3781 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3841 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3901 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     3961 ttggccgatt cattaatgca ggatctcgat cccgcgaaat
//
