LOCUS       sequence-429495-        6795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mitochondrially targeted non-responsive control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6795)
  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 6795)
  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: pAAV.CAG.(mito).cpSFGFP.HaloTag
FEATURES             Location/Qualifiers
     source          1..6795
                     /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     4468..4489
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4592..4613)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    5942..6530
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5995..6015)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6413..6424)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6573..6694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6584..6603)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6638..6657
                     /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 sggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
     3301 cggggcgggg cgaggcggag 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 gatctagcca
     4261 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg
     4321 ccacaacgtg gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     4381 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgtgctgag
     4441 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4501 gatcactctc ggcatggacg agctgtacaa gggcggtacc ggagggagca tgagcaaggg
     4561 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     4621 ccacaagttc agcgtgcgcg gcgagggcga gggcgatgcc accaacggca agctgaccct
     4681 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     4741 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     4801 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcagcttca aggacgacgg
     4861 cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     4921 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcataagc tggagtacaa
     4981 ctttaacctg cagggcggag gctcgggcgg aggtgggtcg ggtggcggcg gatcaatggc
     5041 cgagatcggc accggcttcc ccttcgaccc ccactacgtg gaggtgctgg gcgagcgcat
     5101 gcactacgtt gacgtgggcc cccgcgacgg cacccccgtg ctgttcctgc acggcaaccc
     5161 caccagcagc tacgtgtggc gcaacatcat cccccacgtg gcccccaccc accgcactat
     5221 cgcccccgac ctgatcggca tgggcaagag cgacaagccc gacctgggct acttcttcga
     5281 cgaccacgtg cgcttcatgg acgccttcat cgaggccctg ggcctggagg aggtggtgct
     5341 ggtgatccac gactggggca gcgccctggg cttccactgg gccaagcgca accccgagcg
     5401 cgtgaagggc atcgccttca tggagttcat ccgccccatc cccacctggg acgagtggcc
     5461 cgagttcgcc cgcgagacct tccaggcctt ccgcaccacc gacgtgggcc gcaagctgat
     5521 catcgaccag aacgtgttca tcgagggcac cctgcccatg ggcgtggtgc gccccctgac
     5581 cgaggtggag atggaccact accgcgagcc cttcctgaac cccgtggacc gcgagcccct
     5641 gtggcgcttc cccaacgagc tgcccatcgc cggcgagccc gccaacatcg tggccctggt
     5701 ggaggagtac atggactggc tgcaccagag ccccgtgccc aagctgctgt tctggggcac
     5761 ccccggcgtg ctgatccccc ccgccgaggc cgcccgcctg gccaagagcc tgcccaacgt
     5821 taaggccgtg gacatcggcc ccggcctgaa cctgctccag gaggacaacc ccgacctgat
     5881 cggcagcgag atcgcccgct ggctgagcac cctcgagatc agcggctaga agcttatcga
     5941 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     6001 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     6061 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     6121 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     6181 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     6241 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6301 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     6361 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6421 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6481 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     6541 tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     6601 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     6661 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     6721 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     6781 tggtaaaatc gataa
//
