LOCUS       sequence-72240-e        4103 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial expression vector of N-terminal His-tagged PercevalHR, a 
            fluorescent sensor of ATP-to-ADP ratio..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4103)
  AUTHORS   Tantama M, Martinez-Francois JR, Mongeon R, Yellen G
  TITLE     Imaging energy status in live cells with a fluorescent biosensor of 
            the intracellular ATP-to-ADP ratio.
  JOURNAL   Nat Commun. 2013;4:2550. doi: 10.1038/ncomms3550.
  PUBMED    24096541
REFERENCE   2  (bases 1 to 4103)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2013"; volume: "4:2550. doi"; pages: " 
            10.1038/ncomms3550"
COMMENT     Sequence Label: pRsetB-PercevalHR
FEATURES             Location/Qualifiers
     source          1..4103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             106..126
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             328..579
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSFQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     516..537
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(640..661)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(901..920)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1959..1980
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2143..2247
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2248..2316,2317..3108)
                     /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(2466..2485)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3279..3867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3768..3787
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4021..4038
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgaaacatca ccatcaccat
      121 caccatggag ctagcatgaa aaaggtggaa tccatcatca ggcccgaaaa gctggagatc
      181 gttaagaagg ctctctcgga cgctggatat gtgggtatga ccgtctctga agtcaaaggc
      241 tctggcgtcc agggcggcat cttcgagagg taccgaggaa gggtgtactc tgcaggctac
      301 aacagcgaca acgtctacat caccgccgac aagcagaaga acggcatcaa ggccaacttc
      361 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
      421 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cttccagtcc
      481 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      541 gccgccggga tcactctcgg catggacgag ctgtacaagg gcggttccgg aggcatggtg
      601 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      661 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      721 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      781 accaccctgg gctacggtct ccaatgcttc gcccgctacc ccgaccacat gaagcagcac
      841 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      901 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      961 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctt
     1021 gagtacaacg gcaccatagt agatctgatc cctaaggtaa aaattgaact agtggtgaag
     1081 gaggaggacg tcgataacgt gatagacatt atttgcgaaa atgcccgcac aggtaacccc
     1141 ggcgacggta agatatttgt gatcccagtg gagcgagtgg tcagggtgcg aaccaaagag
     1201 gagggagcat ctggtggtgg atccggtggt ggcggtgcat ctggtatgaa aaaggttgaa
     1261 gctattattc gcccagagaa actggagatt gttaagaagg ccctgtccga cgccggctat
     1321 gttggcatga ccgtttccga ggttaaaggc cgcggtgccg gcggtggcga cctgattcct
     1381 aaggtgaaga tcgagctggt tgttaaggag gaagacgttg ataatgttat cgacattatc
     1441 tgcgagaacg cccgcactgg caatcctggt gacggcaaaa tcttcgttat accggtcgag
     1501 cgaattgtgc gagtacgaac caaagaagag ggtgcctccg gcggtggcgg tggctccggt
     1561 ggcgccagcg gcatgaagaa ggttgaggcc atcatccgcc cagagaagct ggaaatcgtt
     1621 aagaaggcct taaacgacga cggctacgtc ggcatgacag ttagcgaggt taaaggtcgc
     1681 ggtgccggtg ggggtgatct gattcctaag gttaaaatcg agctcgttgt taaggaggag
     1741 gacgttgaca acattatcga tatcatctgc gaaaacgctc gcaccggcaa cccaggcgac
     1801 ggcaagattt tcgtcattcc tgttgagaga gtggtgcgtc cccgtacgaa ggaggagggc
     1861 aaggaagcac tgtgactcga ggggcccaag ctgtagtggg ccatcgccct gatagacggt
     1921 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     1981 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     2041 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     2101 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2161 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2221 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2281 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2341 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2401 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2461 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     2521 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2581 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2641 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2701 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2761 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     2821 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2881 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     2941 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3001 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3061 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3121 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3181 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3241 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3301 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3361 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3421 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3481 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3541 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3601 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3661 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3721 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3781 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3841 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3901 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3961 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4021 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     4081 cgttggccga ttcattaatg cag
//
