LOCUS       sequence-273255-        3999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative control for ATP measurement.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3999)
  AUTHORS   Yaginuma H, Kawai S, Tabata KV, Tomiyama K, Kakizuka A, Komatsuzaki 
            T, Noji H, Imamura H
  TITLE     Diversity in ATP concentrations in a single bacterial cell 
            population revealed by quantitative single-cell imaging.
  JOURNAL   Sci Rep. 2014 Oct 6;4:6522. doi: 10.1038/srep06522.
  PUBMED    25283467
REFERENCE   2  (bases 1 to 3999)
  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: "Sci Rep."; 
            date: "2014-10-6"; volume: "4:6522. doi"; pages: " 
            10.1038/srep06522"
COMMENT     Sequence Label: pRSET-QNA
FEATURES             Location/Qualifiers
     source          1..3999
                     /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(1381..1399)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1395..1442
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1539..1994
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1626..1645)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1836..1857
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2020..2124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2125..2193,2194..2985)
                     /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(2343..2362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3156..3744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3645..3664
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3898..3915
                     /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 tgaaaactgt gaaagtgaat ataacaaccc ctgatgggcc agtctacgac gctgatatcg
      241 agatggtgtc cgtgcgggcc gagagtggtg atctcggcat cctccccggt cacattccca
      301 caaaggcccc actgaagatc ggagctgtgc ggctgaagaa ggacggccaa accgagatgg
      361 tcgcagtctc aggcggcact gttgaagtgc ggcctgacca cgttaccatt aatgctcaag
      421 ccgctgaaac agccgaagga atcgacaaag agagagcaga agccgcaaga cagagggccc
      481 aggagcggct gaactctcaa 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 accgatattc
     1261 gccgggccga gctggcactg cagaaggccc tgaacaagct ggacgtggct gggaaggcaa
     1321 actgaaagct tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     1381 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1441 tgctgaaagg aggaactata tccggatctg gcgtaatagc gaagaggccc gcaccgatcg
     1501 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     1561 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     1621 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1681 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     1741 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     1801 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     1861 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     1921 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     1981 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2041 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2101 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2161 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2221 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2281 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2341 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2401 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2461 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2521 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2581 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2641 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2701 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2761 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2821 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2881 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2941 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3001 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3061 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3121 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3181 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3241 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3301 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3361 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3421 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3481 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3541 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3601 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3661 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3721 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3781 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3841 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3901 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     3961 tggccgattc attaatgcag gatctcgatc ccgcgaaat
//
