LOCUS       sequence-260056-        4026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mutant of QUEEN 84BP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4026)
  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 4026)
  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: pRSAN-QUEEN BPEE
FEATURES             Location/Qualifiers
     source          1..4026
                     /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"
     regulatory      183..192
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             549..800
                     /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     737..758
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(864..885)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1125..1144)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1408..1426)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1422..1469
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1566..2021
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1653..1672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1863..1884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2047..2151
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2152..2220,2221..3012)
                     /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(2370..2389)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3183..3771
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3672..3691
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3925..3942
                     /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 gatccaccgg
      181 tcgccaccat ggtgaaaact gtgaaagtga atataacaac ccctgatggg ccagtctacg
      241 acgctgatat cgagatggtg tccgtgcggg ccgagagtgg tgatctcggc atcctccccg
      301 gtcacattcc cacaaaggcc ccactgaaga tcggagctgt gcggctgaag aaggacggcc
      361 aaacagagga agtcgcagtc tcaggcggca ctgttgaagt gcggcctgac cacgttacca
      421 ttaatgctca agccgctgaa cgggcggagg acattgacgt cctccgcgcc aaagcggcga
      481 aagagcgggc ggagcgccgc ctgcaaagcc agacccgtta caacagccac aacgtctata
      541 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
      601 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
      661 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
      721 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
      781 gcatggacga gctgtacaag ggtggcaccg gagggagcat ggtgagcaag ggcgaggagc
      841 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      901 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      961 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1021 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1081 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1141 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1201 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacctgggtg
     1261 acatcgactt caaacgggcc gaactggcgt taaaacgcgc catgaaccgt ttgagcgttg
     1321 cggaagaaaa gtgagcggcc gcgactctag ataagcttga tccggctgct aacaaagccc
     1381 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     1441 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatctggcg
     1501 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1561 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1621 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1681 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1741 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     1801 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     1861 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     1921 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     1981 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     2041 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2101 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2161 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2221 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2281 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2341 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2401 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2461 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2521 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2581 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2641 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2701 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2761 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2821 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2881 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2941 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3001 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3061 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3121 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3181 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3241 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3301 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3361 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3421 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3481 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3541 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3601 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3661 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3721 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3781 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3841 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3901 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     3961 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcaggat ctcgatcccg
     4021 cgaaat
//
