LOCUS       sequence-493835-        7189 bp    DNA     circular SYN 08-JAN-2026
DEFINITION  iATPSnFR2.S29W.A95K variant with non-responsive tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7189)
  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 7189)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 8, 2026 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: pAAV.CAG.(cyto).iATPSnFR2.S29W.A95K.tdTomato
FEATURES             Location/Qualifiers
     source          1..7189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(256..844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(336..355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1015..1806,1807..1875))
                     /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     1638..1657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1876..1980)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2006..2461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2143..2164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2355..2374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2533..2662
                     /label=AAV2 ITR
     promoter        2975..3252
                     /label=chicken beta-actin promoter
     primer_bind     3531..3553
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3615..3634
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3668..3677
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4225..4246
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4349..4370)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    6314..6902
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6367..6387)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6785..6796)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6945..7066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6956..6975)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7010..7029
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
       61 ggccactccc tctctgcgcg ctcgctcgct cactgactcg ctgcgctcgg tcgttcggct
      121 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      181 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      421 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      661 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1981 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     2041 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2101 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2161 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2221 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2281 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2341 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2401 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2461 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     2521 ctattacgcc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
     2581 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
     2641 aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta cttatctacg
     2701 tagccatgct ctaggaagat cgtaccattg acgtcaataa tgacgtatgt tcccatagta
     2761 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     2821 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     2881 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     2941 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
     3001 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
     3061 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccrggsgg gggggggggg
     3121 ggsggggggs ggggsggggs grggcggaga ggtgcggcgg cagccaatca gagcggcgcg
     3181 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
     3241 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
     3301 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
     3361 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
     3421 tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg
     3481 gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt
     3541 ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt
     3601 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
     3661 ggatcccgcc accatgggca gatctatgaa gactattcac gtgagtgtcg taactcccga
     3721 cgggcctgta tatgaggatg acgttgaaat ggtgagcgtc aaagcaaaat ggggcgagct
     3781 cggtattctc ccaggccaca ttcccttgaa agctccccta gagatcagtg ccgcacgcct
     3841 gaagaaaggg ggcaaaacac agtatatcgc tgtgtcaggc ggcaacctcg aagtgcggcc
     3901 tgacaaggtg accatctatg cccaggcagc cgagagggct gaggatatcg atgtcctgcg
     3961 cgccaagaaa gccaaggaga gagccgagcg ccgactccaa tcacaggtcc tggtcgggaa
     4021 cgtctatatc accgccgaca agcagaagaa cggcatcaag gcgaacttca agatccgcca
     4081 caacgtggag gacggcagca tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     4141 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg tgctgagcaa
     4201 agaccctaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     4261 cactctcggc atggacgagc tgtacaaggg cggtaccgga gggagcatga gcaagggcga
     4321 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     4381 caagttcagc gtgcgcggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa
     4441 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     4501 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     4561 gtccgccatg cccgaaggct acgtccagga gcgcaccatc agcttcaagg acgacggcac
     4621 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     4681 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt
     4741 tatttgtgtg gacatcgatt ttaagcgcgc cgagctcgcc cttaagcgcg caatgaatag
     4801 gctctcagtt gccgaaatga agctgcaggg cggaggctcg ggcggaggtg ggtcgggtgg
     4861 cggcggatca atggttagca agggtgaaga ggtgattaaa gagtttatgc gttttaaagt
     4921 acgcatggag ggtagcatga acggtcatga gtttgagatc gagggcgaag gcgaaggtcg
     4981 tccgtacgag ggtacgcaga ccgctaaact gaaagtaacc aaaggcggtc ctctgccgtt
     5041 cgcgtgggac atcctgtccc cgcagtttat gtatggttcc aaggcctacg tcaaacaccc
     5101 agccgatatc ccggactaca agaagctgtc tttcccggag ggtttcaagt gggagcgtgt
     5161 aatgaacttt gaggacggtg gcctggttac cgtcacccag gatagctctc tgcaagatgg
     5221 caccctgatc tacaaggtga agatgcgtgg tacgaacttt ccgccggatg gcccggtgat
     5281 gcagaaaaag acgatgggat gggaagcatc taccgagcgc ctgtatccgc gtgacggtgt
     5341 tctgaaaggt gagatccacc aggcgctgaa actgaaggac ggcggtcact acctggttga
     5401 atttaagacg atctatatgg ccaaaaagcc ggttcaactg ccaggttact actacgtgga
     5461 caccaaactg gatatcacgt ctcacaacga agattatacc atcgtcgaac agtatgaacg
     5521 tagcgagggc cgtcaccacc tgttcctggg ccacggtact ggttctacgg gttctggttc
     5581 ctctggcact gcgtcttccg aagacaacaa catggcggtg atcaaagagt tcatgaggtt
     5641 caaagtacgt atggaaggtt ctatgaacgg ccatgagttc gagattgaag gtgagggcga
     5701 gggtcgccca tacgagggca ctcagaccgc aaagctgaaa gttaccaaag gtggcccact
     5761 gccttttgcg tgggatatcc tgtctccgca gttcatgtac ggttccaaag cctacgtaaa
     5821 acacccggca gacatcccgg attacaaaaa gctgtccttc ccagagggct tcaaatggga
     5881 acgtgttatg aatttcgaag atggcggcct ggtcactgtt acgcaggaca gcagcctgca
     5941 agacggtact ctgatttaca aagtgaaaat gcgtggcact aacttcccgc ctgacggtcc
     6001 agtaatgcag aagaaaacta tgggctggga agctagcacc gaacgtctgt acccgcgcga
     6061 cggtgtgctg aaaggcgaga ttcaccaggc cctgaagctg aaggatggtg gtcactatct
     6121 ggttgagttc aaaactatct acatggcaaa aaagcctgta caactgccgg gctattacta
     6181 cgttgacact aaactggaca tcacctccca caacgaggat tatactattg ttgaacagta
     6241 cgagcgttct gaaggtcgcc atcatctgtt tctgtatggt atggacgaac tgtataaata
     6301 gaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     6361 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     6421 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     6481 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     6541 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     6601 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     6661 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     6721 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     6781 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     6841 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     6901 gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat acattgatga
     6961 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     7021 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     7081 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
     7141 cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggcta
//
