LOCUS       sequence-260057-        4029 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 4029)
  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 4029)
  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 BPKK
FEATURES             Location/Qualifiers
     source          1..4029
                     /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"
     regulatory      186..195
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             552..803
                     /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     740..761
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(867..888)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1128..1147)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse 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 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat ggtatggcta
      121 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatccac
      181 cggtcgccac catggtgaaa actgtgaaag tgaatataac aacccctgat gggccagtct
      241 acgacgctga tatcgagatg gtgtccgtgc gggccgagag tggtgatctc ggcatcctcc
      301 ccggtcacat tcccacaaag gccccactga agatcggagc tgtgcggctg aagaaggacg
      361 gccaaacaga gaaggtcgca gtctcaggcg gcactgttga agtgcggcct gaccacgtta
      421 ccattaatgc tcaagccgct gaacgggcgg aggacattga cgtcctccgc gccaaagcgg
      481 cgaaagagcg ggcggagcgc cgcctgcaaa gccagacccg ttacaacagc cacaacgtct
      541 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
      601 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
      661 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc
      721 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
      781 tcggcatgga cgagctgtac aagggtggca ccggagggag catggtgagc aagggcgagg
      841 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      901 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      961 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1021 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1081 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1141 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1201 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacctgg
     1261 gtgacatcga cttcaaacgg gccgaactgg cgttaaaacg cgccatgaac cgtttgagcg
     1321 ttgcggaaaa gaagtgagcg gccgcgactc tagataagct 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 gatctcgatc
     4021 ccgcgaaat
//
