LOCUS       sequence-327064-        7557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector (CMV promoter) of 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 7557)
  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 7557)
  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: "Nat 
            Commun."; date: "2013"; volume: "4:2550. doi"; pages: " 
            10.1038/ncomms3550"
COMMENT     Sequence Label: GW1-PercevalHR
FEATURES             Location/Qualifiers
     source          1..7557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        309..688
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        689..892
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     843..863
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     889..913
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1028..1854
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     CDS             2252..2503
                     /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     2440..2461
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2564..2585)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2825..2844)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3921..4055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3958..3977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4012..4031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4078..4094)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4078..4094)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4091..4113)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4102..4118
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4126..4132,4133..4150,4151..4156))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4308..4325)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4479..5067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4559..4578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5238..6029,6030..6098))
                     /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     5861..5880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6155..6668)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6350..6371)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6562..6581
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6840..6858
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6896..6918)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7018..7037
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7269..7291
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7283..7300
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7284..7300
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gcttatatcg tttacggggg atggcgatag acgactttgg tgacttgggc gattctgtgt
       61 gtcgcaaata tcgcagtttc gatataggtg acagacgata tgaggctata tcgccgatag
      121 aggcgacatc aagctggcac atggccaatg catatcgatc tatacattga atcaatattg
      181 gccattagcc atattattca ttggttatat agcataaatc aatattggct attggccatt
      241 gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc caacattacc
      301 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      361 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      421 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      481 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      541 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      601 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      661 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      721 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      781 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      841 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      901 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg
      961 ggaccgatcc agcctccgcg gccgggaacg gtgcattgga acgcggattc cccgtgccaa
     1021 gagtgacgta agtaccgcct atagagtcta taggcccacc cccttggctt cttatgcatg
     1081 ctatactgtt tttggcttgg ggtctataca cccccgcttc ctcatgttat aggtgatggt
     1141 atagcttagc ctataggtgt gggttattga ccattattga ccactcccct attggtgacg
     1201 atactttcca ttactaatcc ataacatggc tctttgccac aactctcttt attggctata
     1261 tgccaataca ctgtccttca gagactgaca cggactctgt atttttacag gatggggtct
     1321 catttattat ttacaaattc acatatacaa caccaccgtc cccagtgccc gcagttttta
     1381 ttaaacataa cgtgggatct ccacgcgaat ctcgggtacg tgttccggac atgggctctt
     1441 ctccggtagc ggcggagctt ctacatccga gccctgctcc catgcctcca gcgactcatg
     1501 gtcgctcggc agctccttgc tcctaacagt ggaggccaga cttaggcaca gcacgatgcc
     1561 caccaccacc agtgtgccgc acaaggccgt ggcggtaggg tatgtgtctg aaaatgagct
     1621 cggggagcgg gcttgcaccg ctgacgcatt tggaagactt aaggcagcgg cagaagaaga
     1681 tgcaggcagc tgagttgttg tgttctgata agagtcagag gtaactcccg ttgcggtgct
     1741 gttaacggtg gagggcagtg tagtctgagc agtactcgtt gctgccgcgc gcgccaccag
     1801 acataatagc tgacagacta acagactgtt cctttccatg ggtcttttct gcagtcaccg
     1861 tccttgacac gaagcttccc ggccacagca atcgatccat gacacaacca gtgacaagca
     1921 gtcccccagt gaaaccgcat ccgcatccga gtccgatacc gataaagatt ctgaatcgga
     1981 gtgagtgccg cgtccgagag cgttccctgt ccacgtccac catcggcgga gcaggtgtgc
     2041 ctgaggcctc tagaccacca tgaaaaaggt tgaatccatc atcaggcccg aaaagctgga
     2101 gatcgttaag aaggctctct cggacgctgg atatgtgggt atgaccgtct ctgaagtcaa
     2161 aggctctggc gtccagggcg gcatcttcga gaggtaccga ggaagggtgt actctgcagg
     2221 ctacaacagc gacaacgtct acatcaccgc cgacaagcag aagaacggca tcaaggccaa
     2281 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca
     2341 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcttcca
     2401 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     2461 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggtt ccggaggcat
     2521 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2581 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2641 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2701 cgtgaccacc ctgggctacg gtctccaatg cttcgcccgc taccccgacc acatgaagca
     2761 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2821 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2881 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2941 gcttgagtac aacggcacca tagtagatct gatccctaag gtaaaaattg aactagtggt
     3001 gaaggaggag gacgtcgata acgtgataga cattatttgc gaaaatgccc gcacaggtaa
     3061 ccccggcgac ggtaagatat ttgtgatccc agtggagcga gtggtcaggg tgcgaaccaa
     3121 agaggaggga gcatctggtg gtggatccgg tggtggcggt gcatctggta tgaaaaaggt
     3181 tgaagctatt attcgcccag agaaactgga gattgttaag aaggccctgt ccgacgccgg
     3241 ctatgttggc atgaccgttt ccgaggttaa aggccgcggt gccggcggtg gcgacctgat
     3301 tcctaaggtg aagatcgagc tggttgttaa ggaggaagac gttgataatg ttatcgacat
     3361 tatctgcgag aacgcccgca ctggcaatcc tggtgacggc aaaatcttcg ttataccggt
     3421 cgagcgaatt gtgcgagtac gaaccaaaga agagggtgcc tccggcggtg gcggtggctc
     3481 cggtggcgcc agcggcatga agaaggttga ggccatcatc cgcccagaga agctggaaat
     3541 cgttaagaag gccttaaacg acgacggcta cgtcggcatg acagttagcg aggttaaagg
     3601 tcgcggtgcc ggtgggggtg atctgattcc taaggttaaa atcgagctcg ttgttaagga
     3661 ggaggacgtt gacaacatta tcgatatcat ctgcgaaaac gctcgcaccg gcaacccagg
     3721 cgacggcaag attttcgtca ttcctgttga gagagtggtg cgtccccgta cgaaggagga
     3781 gggcaaggaa gcactgtgac tcgacgaatt cattgatcat aatcagccat accacatttg
     3841 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3901 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3961 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4021 ccaaactcat caatgtatct tatcatgtct ggatccgtac cgagctcgcg taatcatgtc
     4081 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4141 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactacatt aattgcgttg
     4201 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4261 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4321 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4381 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4441 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4501 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4561 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4621 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4681 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4741 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4801 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4861 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4921 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4981 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5041 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5101 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5161 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5221 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5281 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5341 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5401 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5461 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5521 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5581 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5641 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5701 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5761 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5821 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5881 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5941 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6001 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6061 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaataagcg
     6121 gccgcggcca tgccggccac tagtctcgag ggccaccccg gttgataatc agaaaagccc
     6181 caaaaacagg aagattgtat aagcaaatat ttaaattgta agcgttaata ttttgttaaa
     6241 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6301 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     6361 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     6421 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     6481 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6541 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     6601 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6661 gggcgcgtgg ggatccctcg agttattatt gaagcattta tcagggttat tgtctcatga
     6721 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6781 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6841 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6901 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6961 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     7021 catcagagca gattgtactg agagtgcacc atatgaagac gtcgcctcct cactacttct
     7081 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagg
     7141 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     7201 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     7261 agggttttcc cagtcacgac gttgtaaaac gacggccagt gcccgtgtca aggacggtga
     7321 ctgcagtgaa taataaaatg tgtgtttgtc cgaaatacgc gttttgagat ttctgtcgcc
     7381 gactaaattc atgtcgcgcg atagtggtgt ttatcgccga tagagatggc gatattggaa
     7441 aaatcgatat ttgaaaatat ggcatattga aaatgtcgcc gatgtgagtt tctgtgtaac
     7501 tgatatcgcc atttttccaa aagtgatttt tgggcatacg cgatatctgg cgatagc
//
