LOCUS       sequence-499680-        8218 bp    DNA     circular SYN 10-FEB-2026
DEFINITION  Mammalian expression of photoactivatable GFP targeted to the 
            mitochondrial outer membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8218)
  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 8218)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 10, 2026 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-PAGFPomp25
FEATURES             Location/Qualifiers
     source          1..8218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    147..504
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             569..985
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     915..937
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     953..969
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    995..1369
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1307..1323
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     enhancer        1470..1773
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1775..2051
                     /label=chicken beta-actin promoter
     intron          2052..3068
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2992..3014
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3076..3095
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3168..3177
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3174..3176,3177..3179,3180..3890)
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3219..3240)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3480..3499)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3827..3848
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4088..4676
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4141..4161)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4559..4570)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4679..4695)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4680..4696)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5013..5482
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5612..5629)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5783..6371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5863..5882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6531..7322,7323..7391))
                     /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     7154..7173
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        7543..7922
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7923..8126
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8077..8097
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
       61 acccgtcagc gggggtcttt catttggggg ctcgtccggg atcgggagac ccctgcccag
      121 ggaccaccga cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat
      181 tgtctagtgt ctatgactga ttttatgcgc ctgcgtcggt actagttagc taactagctc
      241 tgtatctggc ggacccgtgg tggaactgac gagttcggaa cacccggccg caaccctggg
      301 agacgtccca gggacttcgg gggccgtttt tgtggcccga cctgagtcca aaaatcccga
      361 tcgttttgga ctctttggtg cacccccctt agaggaggga tatgtggttc tggtaggaga
      421 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt gggaccgaag
      481 ccgcgccgcg cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
      541 tctgtatttg tctgaaaata tgggcccggg ccagactgtt accactccct taagtttgac
      601 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
      661 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
      721 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
      781 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
      841 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
      901 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
      961 agccctcact ccttctctag gcgcccccat atggccatat gagatcttat atggggcacc
     1021 cccgcccctt gtaaacttcc ctgaccctga catgacaaga gttactaaca gcccctctct
     1081 ccaagctcac ttacaggctc tctacttagt ccagcacgaa gtctggagac ctctggcggc
     1141 agcctaccaa gaacaactgg accgaccggt ggtacctcac ccttaccgag tcggcgacac
     1201 agtgtgggtc cgccgacacc agactaagaa cctagaacct cgctggaaag gaccttacac
     1261 agtcctgctg accaccccca ccgccctcaa agtagacggc atcgcagctt ggatacacgc
     1321 cgcccacgtg aaggctgccg accccggggg tggaccatcc tctagactgc cggatcccag
     1381 tgtggtggta cgggaattct gcaggaatct agttattaat agtaatcaat tacggggtca
     1441 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     1501 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     1561 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     1621 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     1681 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     1741 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
     1801 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
     1861 ttttgtgcag cgatgggggc gggggggggg gggggcgcgc gccaggcggg gcggggcggg
     1921 gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc
     1981 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg
     2041 cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct
     2101 cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg
     2161 gcccttctcc tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt
     2221 ggctgcgtga aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg
     2281 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg
     2341 gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga
     2401 gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg
     2461 tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac
     2521 cccccctgca cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc
     2581 gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc
     2641 cgggcggggc ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc
     2701 ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt
     2761 gcgagagggc gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc
     2821 gccgccgcac cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa
     2881 tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc
     2941 ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct
     3001 tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt
     3061 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
     3121 tgatttgata ccgcgggccc gggatccctc gactagcgct accggtcgcc accatggtga
     3181 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     3241 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     3301 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     3361 ccaccttcag ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     3421 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     3481 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     3541 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     3601 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     3661 aggccaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     3721 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     3781 gccaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     3841 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tccggccgga
     3901 ctcagatctc gagctcaagc ttcgaattcc atcgaggtga aggagagcca agtggagttc
     3961 ctgtagctat ggtgctgcta ccagtgtttg ccctcaccat ggtagcagtt tgggccttcg
     4021 tgagataccg aaagcagctt tgaggatcga ggccgcgact ctagaattcg atatcaagct
     4081 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4141 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4201 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4261 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4321 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4381 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4441 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     4501 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4561 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4621 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     4681 ataccgtcga cctcgatcga ggaattccag ctgagcgccg gtcgctacca ttaccagttg
     4741 gtctggtgtc aaaaataata ataaccgggc aggccatgtc tgcccgtatt tcgcgtaagg
     4801 aaatccatta tgtactattt tcgagcggcc gccagcacag tggtcgacga taaaataaaa
     4861 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     4921 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     4981 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     5041 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     5101 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     5161 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     5221 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     5281 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     5341 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     5401 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     5461 acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct
     5521 taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag
     5581 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5641 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5701 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5761 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5821 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5881 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5941 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6001 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6061 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6121 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6181 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6241 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6301 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6361 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6421 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6481 tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6541 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6601 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6661 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6721 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6781 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6841 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6901 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6961 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7021 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7081 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7141 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7201 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7261 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7321 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7381 tgaatactca tactcttcct ttttcaatga tccggccatt agccatatta ttcattggtt
     7441 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcca tatcataata
     7501 tgtacattta tattggctca tgtccaacat taccgccatg ttgacattga ttattgacta
     7561 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     7621 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     7681 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     7741 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     7801 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     7861 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     7921 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     7981 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     8041 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     8101 ggtgggaggt ctatataagc agagctcaat aaaagagccc acaacccctc actcggcgcg
     8161 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgc
//
