LOCUS       sequence-366323-        8158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of tagRFP targeted to the mitochondrial matrix.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8158)
  AUTHORS   Mishra P, Carelli V, Manfredi G, Chan DC
  TITLE     Proteolytic cleavage of Opa1 stimulates mitochondrial inner membrane
            fusion and couples fusion to oxidative phosphorylation.
  JOURNAL   Cell Metab. 2014 Apr 1;19(4):630-41. doi: 
            10.1016/j.cmet.2014.03.011.
  PUBMED    24703695
REFERENCE   2  (bases 1 to 8158)
  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"; doi: 
            "10.1016/j.cmet.2014.03"; journalName: "Cell Metab"; date: 
            "2014-04-1- 1"; volume: "19"; issue: "4"; pages: "630-41"
COMMENT     Sequence Label: pclbw-mitoTagRFP
FEATURES             Location/Qualifiers
     source          1..8158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          110..1126
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1050..1072
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1134..1153
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1214..1238
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1245..1319
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1341..2069
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTDM
                     ALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVA
                     RYCDLPSKLGHKLNGMDELYK"
     protein_bind    complement(2073..2097)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2142..2730
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2195..2215)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2613..2624)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2733..2749)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2734..2750)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3067..3536
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3666..3683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3837..4425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3917..3936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4585..5376,5377..5445))
                     /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     5208..5227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        5597..5976
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5977..6180
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6131..6151
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    6419..6776
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6841..7257
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7187..7209
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7225..7241
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    7267..7641
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     7579..7595
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     enhancer        7742..8045
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
       61 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
      121 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
      181 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
      241 ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg
      301 gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg
      361 cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg
      421 cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc
      481 gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg
      541 gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg
      601 agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct
      661 cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg
      721 ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg
      781 cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc
      841 tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg
      901 gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc
      961 gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg
     1021 ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc
     1081 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
     1141 gctggttatt gtgctgtctc atcattttgg caaagaattg atttgatacc gcgggcccgg
     1201 gatccctcga atcacaagtt tgtacaaaaa agcaggctga caccatgtcc gtcctgacgc
     1261 cgctgctgct gcggggcttg acaggctcgg cccggcggct cccagtgccg cgcgccaaga
     1321 tccattcgtt gggggatccg gtgtctaagg gcgaagagct gattaaggag aacatgcaca
     1381 tgaagctgta catggagggc accgtgaaca accaccactt caagtgcaca tccgagggcg
     1441 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc
     1501 tccccttcgc cttcgacatc ctggctacca gcttcatgta cggcagcaga accttcatca
     1561 accacaccca gggcatcccc gatttcttta agcagtcctt ccctgagggc ttcacatggg
     1621 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc
     1681 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttccca tccaacggcc
     1741 ctgtgatgca gaagaaaaca ctcggctggg aggccaacac cgagatgctg taccccgctg
     1801 acggcggcct ggaaggcaga accgacatgg ccctgaagct cgtgggcggg ggccacctga
     1861 tctgcaactt caagaccaca tacagatcca agaaacccgc taagaacctc aagatgcccg
     1921 gcgtctacta tgtggaccac agactggaaa gaatcaagga ggccgacaaa gagacctacg
     1981 tcgagcagca cgaggtggct gtggccagat actgcgacct ccctagcaaa ctggggcaca
     2041 aacttaatgg catggacgag ctgtacaagt aaacccagct ttcttgtaca aagtggtgat
     2101 tcgaggccgc gactctagaa ttcgatatca agcttatcga taatcaacct ctggattaca
     2161 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2221 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2281 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     2341 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     2401 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     2461 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     2521 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     2581 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     2641 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     2701 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga tcgaggaatt
     2761 ccagctgagc gccggtcgct accattacca gttggtctgg tgtcaaaaat aataataacc
     2821 gggcaggcca tgtctgcccg tatttcgcgt aaggaaatcc attatgtact attttcgagc
     2881 ggccgccagc acagtggtcg acgataaaat aaaagatttt atttagtctc cagaaaaagg
     2941 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     3001 aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg
     3061 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     3121 ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     3181 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt
     3241 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     3301 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     3361 ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg
     3421 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     3481 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcacatg
     3541 cagcatgtat caaaattaat ttggtttttt ttcttaagta tttacattaa atggccatag
     3601 ttgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3661 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3721 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3781 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3841 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3901 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3961 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4021 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4081 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4141 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4201 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4261 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4321 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4381 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4441 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4501 agattatcaa aaaggatctt cacctagatc cttttgcggc cgcaaatcaa tctaaagtat
     4561 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4621 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4681 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4741 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4801 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4861 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4921 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4981 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5041 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5101 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5161 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5221 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5281 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5341 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5401 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5461 atgatccggc cattagccat attattcatt ggttatatag cataaatcaa tattggctat
     5521 tggccattgc atacgttgta tccatatcat aatatgtaca tttatattgg ctcatgtcca
     5581 acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     5641 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     5701 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     5761 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     5821 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     5881 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     5941 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     6001 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     6061 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     6121 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     6181 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgattga ctgagtcgcc
     6241 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     6301 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatttggg
     6361 ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac cgggaggtaa
     6421 gctggccagc aacttatctg tgtctgtccg attgtctagt gtctatgact gattttatgc
     6481 gcctgcgtcg gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg
     6541 acgagttcgg aacacccggc cgcaaccctg ggagacgtcc cagggacttc gggggccgtt
     6601 tttgtggccc gacctgagtc caaaaatccc gatcgttttg gactctttgg tgcacccccc
     6661 ttagaggagg gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg
     6721 tctgaatttt tgctttcggt ttgggaccga agccgcgccg cgcgtcttgt ctgctgcagc
     6781 atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa tatgggcccg
     6841 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
     6901 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
     6961 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
     7021 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
     7081 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
     7141 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
     7201 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccccc
     7261 atatggccat atgagatctt atatggggca cccccgcccc ttgtaaactt ccctgaccct
     7321 gacatgacaa gagttactaa cagcccctct ctccaagctc acttacaggc tctctactta
     7381 gtccagcacg aagtctggag acctctggcg gcagcctacc aagaacaact ggaccgaccg
     7441 gtggtacctc acccttaccg agtcggcgac acagtgtggg tccgccgaca ccagactaag
     7501 aacctagaac ctcgctggaa aggaccttac acagtcctgc tgaccacccc caccgccctc
     7561 aaagtagacg gcatcgcagc ttggatacac gccgcccacg tgaaggctgc cgaccccggg
     7621 ggtggaccat cctctagact gccggatccc agtgtggtgg tacgggaatt ctgcaggaat
     7681 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     7741 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     7801 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     7861 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     7921 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     7981 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     8041 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
     8101 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggg
//
