LOCUS       sequence-469930-        8385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cytosolic low affinity ratiometric ATP sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8385)
  AUTHORS   Marvin JS, Kokotos AC, Kumar M, Pulido C, Tkachuk AN, Yao JS, Brown 
            TA, Ryan TA
  TITLE     iATPSnFR2: A high-dynamic-range fluorescent sensor for monitoring 
            intracellular ATP.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 May 21;121(21):e2314604121. doi: 
            10.1073/pnas.2314604121. Epub 2024 May 15.
  PUBMED    38748581
REFERENCE   2  (bases 1 to 8385)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2024-05-21"; volume: "121(21):e2314604121. 
            doi"; pages: " 10.1073/pnas.2314604121. Epub 2024 May 15"
COMMENT     Sequence Label: plenti.CMV.(cyto).iATPSnFR2.A95A.A119L.miRFP67...
FEATURES             Location/Qualifiers
     source          1..8385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1804..1920
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1943..2246
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2247..2450
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2401..2421
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2447..2471
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     3082..3103
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3206..3227)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    4231..4819
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4284..4304)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4702..4713)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4906..5139
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5211..5332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5248..5267)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5302..5321
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5372..5507
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5527..5546)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5528..5546)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5556..5573)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5556..5572)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5565..5587)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5714..6169
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5801..5820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6011..6032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6195..6299
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6300..6368,6369..7160)
                     /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(6518..6537)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7331..7919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7820..7839
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8073..8090
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8207..8228
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8243..8248,8249..8266,8267..8273)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8281..8297
                     /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)."
     primer_bind     8286..8308
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8305..8321
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8305..8321
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8340..8360
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8342..8360
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgataagc ttgggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     1981 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     2041 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     2101 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     2161 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     2221 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     2281 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     2341 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     2401 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     2461 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccgcc
     2521 cgggcgtacg ggatccgcca ccatgaagac tattcacgtg agtgtcgtaa ctcccgacgg
     2581 gcctgtatat gaggatgacg ttgaaatggt gagcgtcaaa gcaaaaagtg gcgagctcgg
     2641 tattctccca ggccacattc ccttgaaagc tcccctagag atcagtgccg cacgcctgaa
     2701 gaaagggggc aaaacacagt atatcgctgt gtcaggcggc aacctcgaag tgcggcctga
     2761 caaggtgacc atctatgccc aggcagccga gagggctgag gatatcgatg tcctgcgcgc
     2821 caaggccgcc aaggagagag ccgagcgccg actccaatca caggtcctgg tcgggaacgt
     2881 ctatatcacc gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
     2941 cgtggaggac ggcagcatgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     3001 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgtgc tgagcaaaga
     3061 ccctaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     3121 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatgagca agggcgagga
     3181 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3241 gttcagcgtg cgcggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     3301 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     3361 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3421 cgccatgccc gaaggctacg tccaggagcg caccatcagc ttcaaggacg acggcaccta
     3481 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3541 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactttat
     3601 ttgtgtggac atcgatttta agcgcgccga gctcttgctt aagcgcgcaa tgaataggct
     3661 ctcagttgcc gaaatgaagc tgcagggcgg aggctcgggc ggaggtgggt cgggtggcgg
     3721 cggatcaatg gcaaacctgg acaagatgct gaacaccacc gtgaccgagg tgcgcaagtt
     3781 cctgcaagcg gacagagtgt gcgtgttcaa gttcgaggaa gattactccg gcaccgtctc
     3841 gcacgaagcc gtggacgaca gatggattag catcctgaag acccaggtgc aggacagata
     3901 cttcatggaa accagaggcg aggaatacgt ccacggcaga taccaggcca tcgccgacat
     3961 ctacacagcc aatctggtcg agtgctacag agacctgctg atcgagtttc aggtgcgggc
     4021 cattctggct gtccccatcc tgcaaggcaa gaagctgtgg ggcctgctgg tggcccacca
     4081 actggccggc cctcgggagt ggcagacctg ggaaatcgac ttcctgaaac agcaagccgt
     4141 ggtgatgggc atcgccatcc agcagagcta gacgcgttct agacctgcag ggtcgactcc
     4201 ggatgatcag ggccctgtac agatatcgac aatcaacctc tggattacaa aatttgtgaa
     4261 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     4321 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     4381 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     4441 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     4501 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     4561 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4621 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4681 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     4741 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     4801 ctttgggccg cctccccgcc tggaattcga gctcggtacc tttaagacca atgacttaca
     4861 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     4921 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     4981 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     5041 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     5101 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
     5161 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
     5221 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5281 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5341 ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5401 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5461 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc
     5521 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     5581 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     5641 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     5701 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5761 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5821 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5881 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5941 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6001 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     6061 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6121 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     6181 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6241 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6301 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6361 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6421 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6481 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6541 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6601 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6661 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6721 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6781 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6841 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6901 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6961 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     7021 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7081 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7141 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     7201 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7261 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7321 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7381 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7441 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     7501 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7561 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7621 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7681 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7741 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7801 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7861 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7921 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7981 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     8041 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     8101 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     8161 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     8221 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     8281 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
     8341 caattaaccc tcactaaagg gaacaaaagc tggagctgca agctt
//
