LOCUS       sequence-274823-        8117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CFP targeted to the mitochondrial matrix.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8117)
  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 8117)
  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-mitoCFP
FEATURES             Location/Qualifiers
     source          1..8117
                     /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"
     CDS             1216..1290
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      1318..1327
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1324..1326,1327..1329,1330..2043)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1369..1390)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1630..1649)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1977..1998
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2102..2690
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2155..2175)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2573..2584)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2693..2709)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2694..2710)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3027..3496
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3626..3643)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3797..4385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3877..3896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4545..5336,5337..5405))
                     /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     5168..5187
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        5557..5936
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5937..6140
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6091..6111
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    6379..6736
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6801..7217
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7147..7169
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7185..7201
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    7227..7601
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     7539..7555
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     enhancer        7702..8005
                     /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 ctagcatgtc cgtcctgacg ccgctgctgc tgcggggctt gacaggctcg
     1261 gcccggcggc tcccagtgcc gcgcgccaag atccattcgt tgggggatcc accggtcgcc
     1321 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1381 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1441 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1501 accctcgtga ccaccctgac ctggggcgtg cagtgcttca gccgctaccc cgaccacatg
     1561 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1621 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1681 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1741 cacaagctgg agtacaacta catcagccac aacgtctata tcaccgccga caagcagaag
     1801 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1861 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1921 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1981 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     2041 taaagcggcc gcgactctag tcgaggccgc gactctagaa ttcgatatca agcttatcga
     2101 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2161 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2221 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2281 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2341 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2401 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2461 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     2521 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2581 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2641 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     2701 tcgacctcga tcgaggaatt ccagctgagc gccggtcgct accattacca gttggtctgg
     2761 tgtcaaaaat aataataacc gggcaggcca tgtctgcccg tatttcgcgt aaggaaatcc
     2821 attatgtact attttcgagc ggccgccagc acagtggtcg acgataaaat aaaagatttt
     2881 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     2941 ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag
     3001 atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     3061 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag
     3121 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     3181 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     3241 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     3301 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     3361 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     3421 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     3481 cagcgggggt ctttcacatg cagcatgtat caaaattaat ttggtttttt ttcttaagta
     3541 tttacattaa atggccatag ttgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3601 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3661 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3721 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3781 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3841 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3901 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3961 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4021 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4081 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4141 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4201 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4261 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4321 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4381 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4441 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttgcggc
     4501 cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4561 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4621 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4681 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4741 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4801 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4861 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4921 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4981 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5041 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5101 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5161 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5221 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5281 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5341 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5401 ctcatactct tcctttttca atgatccggc cattagccat attattcatt ggttatatag
     5461 cataaatcaa tattggctat tggccattgc atacgttgta tccatatcat aatatgtaca
     5521 tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt
     5581 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     5641 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     5701 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     5761 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     5821 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     5881 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     5941 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     6001 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     6061 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     6121 aggtctatat aagcagagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
     6181 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     6241 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     6301 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc
     6361 gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt
     6421 gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg
     6481 gcggacccgt ggtggaactg acgagttcgg aacacccggc cgcaaccctg ggagacgtcc
     6541 cagggacttc gggggccgtt tttgtggccc gacctgagtc caaaaatccc gatcgttttg
     6601 gactctttgg tgcacccccc ttagaggagg gatatgtggt tctggtagga gacgagaacc
     6661 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttgggaccga agccgcgccg
     6721 cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     6781 tgtctgaaaa tatgggcccg ggccagactg ttaccactcc cttaagtttg accttaggtc
     6841 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt
     6901 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca
     6961 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg
     7021 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc
     7081 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt
     7141 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca
     7201 ctccttctct aggcgccccc atatggccat atgagatctt atatggggca cccccgcccc
     7261 ttgtaaactt ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc
     7321 acttacaggc tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc
     7381 aagaacaact ggaccgaccg gtggtacctc acccttaccg agtcggcgac acagtgtggg
     7441 tccgccgaca ccagactaag aacctagaac ctcgctggaa aggaccttac acagtcctgc
     7501 tgaccacccc caccgccctc aaagtagacg gcatcgcagc ttggatacac gccgcccacg
     7561 tgaaggctgc cgaccccggg ggtggaccat cctctagact gccggatccc agtgtggtgg
     7621 tacgggaatt ctgcaggaat ctagttatta atagtaatca attacggggt cattagttca
     7681 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     7741 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     7801 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     7861 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     7921 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     7981 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
     8041 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
     8101 agcgatgggg gcggggg
//
