LOCUS       sequence-409925-        7420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of the BRET ATP sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7420)
  AUTHORS   Min SH, French AR, Trull KJ, Tat K, Varney SA, Tantama M
  TITLE     Ratiometric BRET Measurements of ATP with a Genetically-Encoded 
            Luminescent Sensor.
  JOURNAL   Sensors (Basel). 2019 Aug 10;19(16). pii: s19163502. doi: 
            10.3390/s19163502.
  PUBMED    31405152
REFERENCE   2  (bases 1 to 7420)
  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: "Sensors 
            (Basel)."; date: "2019-08-10"; volume: "19(16). pii: s19163502. 
            doi"; pages: " 10.3390/s19163502"
COMMENT     Sequence Label: GW1-ARSeNL
FEATURES             Location/Qualifiers
     source          1..7420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(322..340)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     590..609
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1913..2047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1950..1969)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2004..2023
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2070..2086)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2070..2086)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2083..2105)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2094..2110
                     /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(2118..2124,2125..2142,2143..2148))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2300..2317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2471..3059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2551..2570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3230..4021,4022..4090))
                     /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     3853..3872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4147..4660)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4342..4363)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4554..4573
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4832..4850
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4888..4910)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5010..5029
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5261..5283
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5275..5292
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5276..5292
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5858..6237
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6238..6441
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6392..6412
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6438..6462
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          6577..7403
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
ORIGIN
        1 aagcttcccg gccacagcaa tcgatccatg acacaaccag tgacaagcag tcccccagtg
       61 aaaccgcatc cgcatccgag tccgataccg ataaagattc tgaatcggag tgagtgccgc
      121 gtccgagagc gttccctgtc cacgtccacc atcggcggag caggtgtgcc tgaggcctct
      181 agaccaccat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
      241 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
      301 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
      361 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg
      421 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
      481 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
      541 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
      601 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
      661 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact
      721 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc
      781 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
      841 ccgaaggacg tcattctaca ggccgccata tcgatatgaa aactgtgaaa gtgaatataa
      901 caacccctga tgggccagtc tacgacgctg atatcgagat ggtgtccgtg cgggccgaga
      961 gtggtgatct cggcatcctc cccggtcaca ttcccacaaa ggccccactg aagatcggag
     1021 ctgtgcggct gaagaaggac ggccaaaccg agatggtcgc agtctcaggc ggcactgttg
     1081 aagtgcggcc tgaccacgtt accattaatg ctcaagccgc tgaaacagcc gaaggaatcg
     1141 acaaagagag agcagaagcc gcaagacaga gggcccagga gcggctgaac tctcaatccg
     1201 atgacaccga tattcgccgg gccgagctgg cactgcagag ggccctgaac agactggacg
     1261 tggctgggaa ggcaaacgaa ttcatgctgg aggactttgt tggggactgg cgacagacag
     1321 ccggctacaa cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc
     1381 tcggggtgtc cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga
     1441 tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg
     1501 aaaaaatttt taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact
     1561 atggcacact ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt
     1621 atgaaggcat cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg
     1681 gcaacaaaat tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa
     1741 ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat tctggcctaa actcgacgaa
     1801 ttcattgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     1861 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     1921 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1981 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     2041 ctggatccgt accgagctcg cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     2101 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2161 taatgagtga gctaactaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2221 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2281 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2341 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2401 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2461 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2521 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2581 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2641 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2701 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2761 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2821 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2881 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2941 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3001 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3061 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3121 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3181 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3241 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3301 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3361 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3421 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3481 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3541 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3601 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3661 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3721 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3781 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3841 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3901 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3961 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4021 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4081 gaatactcat actcttcctt tttcaataag cggccgcggc catgccggcc actagtctcg
     4141 agggccaccc cggttgataa tcagaaaagc cccaaaaaca ggaagattgt ataagcaaat
     4201 atttaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4261 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4321 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4381 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4441 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4501 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4561 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4621 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ggggatccct cgagttatta
     4681 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4741 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     4801 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     4861 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     4921 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     4981 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5041 ccatatgaag acgtcgcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     5101 ggcctctgca taaataaaaa aaattagtca gggcgccatt cgccattcag gctgcgcaac
     5161 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     5221 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     5281 acgacggcca gtgcccgtgt caaggacggt gactgcagtg aataataaaa tgtgtgtttg
     5341 tccgaaatac gcgttttgag atttctgtcg ccgactaaat tcatgtcgcg cgatagtggt
     5401 gtttatcgcc gatagagatg gcgatattgg aaaaatcgat atttgaaaat atggcatatt
     5461 gaaaatgtcg ccgatgtgag tttctgtgta actgatatcg ccatttttcc aaaagtgatt
     5521 tttgggcata cgcgatatct ggcgatagcg cttatatcgt ttacggggga tggcgataga
     5581 cgactttggt gacttgggcg attctgtgtg tcgcaaatat cgcagtttcg atataggtga
     5641 cagacgatat gaggctatat cgccgataga ggcgacatca agctggcaca tggccaatgc
     5701 atatcgatct atacattgaa tcaatattgg ccattagcca tattattcat tggttatata
     5761 gcataaatca atattggcta ttggccattg catacgttgt atccatatca taatatgtac
     5821 atttatattg gctcatgtcc aacattaccg ccatgttgac attgattatt gactagttat
     5881 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     5941 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     6001 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     6061 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     6121 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     6181 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     6241 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     6301 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     6361 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     6421 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
     6481 cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg
     6541 tgcattggaa cgcggattcc ccgtgccaag agtgacgtaa gtaccgccta tagagtctat
     6601 aggcccaccc ccttggcttc ttatgcatgc tatactgttt ttggcttggg gtctatacac
     6661 ccccgcttcc tcatgttata ggtgatggta tagcttagcc tataggtgtg ggttattgac
     6721 cattattgac cactccccta ttggtgacga tactttccat tactaatcca taacatggct
     6781 ctttgccaca actctcttta ttggctatat gccaatacac tgtccttcag agactgacac
     6841 ggactctgta tttttacagg atggggtctc atttattatt tacaaattca catatacaac
     6901 accaccgtcc ccagtgcccg cagtttttat taaacataac gtgggatctc cacgcgaatc
     6961 tcgggtacgt gttccggaca tgggctcttc tccggtagcg gcggagcttc tacatccgag
     7021 ccctgctccc atgcctccag cgactcatgg tcgctcggca gctccttgct cctaacagtg
     7081 gaggccagac ttaggcacag cacgatgccc accaccacca gtgtgccgca caaggccgtg
     7141 gcggtagggt atgtgtctga aaatgagctc ggggagcggg cttgcaccgc tgacgcattt
     7201 ggaagactta aggcagcggc agaagaagat gcaggcagct gagttgttgt gttctgataa
     7261 gagtcagagg taactcccgt tgcggtgctg ttaacggtgg agggcagtgt agtctgagca
     7321 gtactcgttg ctgccgcgcg cgccaccaga cataatagct gacagactaa cagactgttc
     7381 ctttccatgg gtcttttctg cagtcaccgt ccttgacacg
//
