LOCUS       sequence-259260-        3998 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 3998)
  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 3998)
  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-QUEEN 7mu E83A
FEATURES             Location/Qualifiers
     source          1..3998
                     /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             111..143
                     /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(147..155,156..170)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             534..785
                     /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     722..743
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(849..870)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1110..1129)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1380..1398)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1394..1441
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1538..1993
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1625..1644)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1835..1856
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2019..2123
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2124..2192,2193..2984)
                     /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(2342..2361)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3155..3743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3644..3663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3897..3914
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatggt atggctagca
      121 tgactggtgg acagcaaatg ggtcgggatc tgtacgacga tgacgataag gatccgatga
      181 aaacgatcca cgtgagcgtc actactcctg atggcccggt gtacgaagac gatgttgaaa
      241 tggtcagcgt caaagcgaaa agcggcgagc tcggcatttt gccggggcac attccgctta
      301 aggccccgct cgagatcagc gcggcccggc tgaaaaaagg cggcaaaacg caatacattg
      361 ccgtcagcgg cggcaatttg gaagtccgcc cggacaaagt gacgattaac gctcaagctg
      421 ctgctcgggc ggaggacatt gacgtcctcc gcgccaaagc ggcgaaagag cgggcggagc
      481 gccgcctgca aagccagacc cgttacaaca gccacaacgt ctatatcatg gccgacaagc
      541 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      601 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      661 acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
      721 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      781 acaagggtgg caccggaggg agcatggtga gcaagggcga ggagctgttc accggggtgg
      841 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      901 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      961 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1021 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1081 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1141 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1201 aggacggcaa catcctgggg cacaagctgg agtacaacct gggtgacatc gacttcaaac
     1261 gggccgaact ggcgttaaaa cgcgccatga accgtttgag cgttgcggaa atgaagtgaa
     1321 ttcgaagctt gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     1381 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1441 gctgaaagga ggaactatat ccggatctgg cgtaatagcg aagaggcccg caccgatcgc
     1501 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     1561 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     1621 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     1681 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     1741 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     1801 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     1861 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     1921 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     1981 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2041 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2101 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2161 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2221 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2281 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2341 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2401 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2461 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2521 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2581 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2641 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2701 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2761 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2821 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2881 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2941 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3001 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3061 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3121 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3181 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3241 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3301 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3361 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3421 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3481 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3541 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3601 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3661 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3721 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3781 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3841 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3901 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     3961 ggccgattca ttaatgcagg atctcgatcc cgcgaaat
//
