LOCUS       sequence-431762-        7200 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses mitochondrially targeted low affinity ratiometric ATP 
            sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7200)
  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 7200)
  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: "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.(mito).iATPSnFR2.A95A.A119L.HaloTag
FEATURES             Location/Qualifiers
     source          1..7200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   61..190
                     /label=AAV2 ITR
     rep_origin      complement(427..1015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(507..526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1186..1977,1978..2046))
                     /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     1809..1828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2047..2151)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2177..2632)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2314..2335)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2526..2545
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2704..2833
                     /label=AAV2 ITR
     promoter        3146..3423
                     /label=chicken beta-actin promoter
     primer_bind     3702..3724
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3786..3805
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3845..3919
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             3950..4021
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             4052..4123
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             4151..4222
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     4801..4822
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4925..4946)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             5450..6331
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEIS"
     misc_feature    6347..6935
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6400..6420)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6818..6829)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6978..7099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6989..7008)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7043..7062
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
       61 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
      121 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
      181 gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      841 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2161 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2221 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2281 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2341 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2401 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2461 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2521 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2581 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     2641 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     2701 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     2761 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     2821 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
     2881 tctaggaaga tcgtaccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2941 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     3001 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     3061 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     3121 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
     3181 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
     3241 gcgatggggg cggggggggg gggggggcgc gcgccrgggg ggggggggsg gggsgggggg
     3301 sggggsgggg sgrggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
     3361 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
     3421 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
     3481 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
     3541 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
     3601 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggctgtcc
     3661 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     3721 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     3781 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tggatcccgc
     3841 caccatgtct gtgctgacac ctctgctgct gagaggcctg acaggctctg ctagaaggct
     3901 gcctgtgcct agggccaaga tccactctct gccacctgaa ggaccttgga gcgtgctgac
     3961 tccactgctc ttgaggggac tcactggatc tgctcggaga ctgccagtgc cacgcgctaa
     4021 gattcatagc ttgccgcctg aaggcggctc cagtgttctg acaccactgt tgctcagggg
     4081 attgactggc agcgcacgta gactccctgt gccaagagcc aaaattcaca gtctcccacc
     4141 ggaaggcggt tccgtcctga cacctttgtt gttgcgcggc ttgactggct cagcaagacg
     4201 gctgccagtt ccaagagcaa agatccacag cctgcctcca gagggcccta gatctggaag
     4261 catgaagact attcacgtga gtgtcgtaac tcccgacggg cctgtatatg aggatgacgt
     4321 tgaaatggtg agcgtcaaag caaaaagtgg cgagctcggt attctcccag gccacattcc
     4381 cttgaaagct cccctagaga tcagtgccgc acgcctgaag aaagggggca aaacacagta
     4441 tatcgctgtg tcaggcggca acctcgaagt gcggcctgac aaggtgacca tctatgccca
     4501 ggcagccgag agggctgagg atatcgatgt cctgcgcgcc aaggccgcca aggagagagc
     4561 cgagcgccga ctccaatcac aggtcctggt cgggaacgtc tatatcaccg ccgacaagca
     4621 gaagaacggc atcaaggcga acttcaagat ccgccacaac gtggaggacg gcagcatgca
     4681 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     4741 caaccactac ctgagcaccc agtccgtgct gagcaaagac cctaacgaga agcgcgatca
     4801 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     4861 caagggcggt accggaggga gcatgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     4921 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgc gcggcgaggg
     4981 cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     5041 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     5101 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     5161 ccaggagcgc accatcagct tcaaggacga cggcacctac aagacccgcg ccgaggtgaa
     5221 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     5281 cggcaacatc ctggggcaca agctggagta caactttatt tgtgtggaca tcgattttaa
     5341 gcgcgccgag ctcttgctta agcgcgcaat gaataggctc tcagttgccg aaatgaagct
     5401 gcagtctacc ggctccggaa atgctgtggg ccaggacacg caggagcgtg aaatcggcac
     5461 cggcttccct ttcgaccccc actacgtgga agtgctgggc gagagaatgc actatgtgga
     5521 cgtgggccct agggacggaa cccccgtgct gtttctgcac ggcaacccca ccagcagcta
     5581 cgtgtggcgg aacatcatcc cccacgtggc ccccacccac agatgtatcg cccctgacct
     5641 gatcggcatg ggcaagagcg acaagcccga cctgggctac ttcttcgacg accacgtgcg
     5701 gttcatggac gccttcatcg aggccctggg cctggaagag gtggtgctcg tgatccacga
     5761 ttggggctct gccctgggct tccactgggc caagagaaac cccgagagag tgaagggaat
     5821 cgccttcatg gaattcatca gacccatccc cacctgggac gagtggcccg agttcgccag
     5881 agagacattc caggccttca gaaccaccga cgtgggcaga aagctgatca tcgaccagaa
     5941 cgtgttcatc gaaggcaccc tgcctatggg cgtcgtgcgg cctctgaccg aggtggaaat
     6001 ggaccactac agagagccct ttctgaaccc cgtggaccgc gaacctctgt ggcggttccc
     6061 taacgagctg cctatcgctg gcgagcccgc caacattgtg gccctggtgg aagagtacat
     6121 ggactggctg caccagagcc ccgtgcccaa gctgctgttt tggggaacac ccggcgtgct
     6181 gatccctcct gctgaggctg ctagactggc caagagcctg cccaactgca aggccgtgga
     6241 tatcggccct ggcctgaacc tgctgcagga agataacccc gatctgatcg gctccgagat
     6301 cgcccggtgg ctgagcaccc tggaaatcag ctagaagctt atcgataatc aacctctgga
     6361 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     6421 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     6481 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     6541 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     6601 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     6661 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     6721 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     6781 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     6841 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     6901 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac ccgggcggcc
     6961 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     7021 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     7081 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     7141 ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
//
