LOCUS       Exported                4759 bp ds-DNA     circular SYN 20-DEC-2021
DEFINITION  Expresses a mitochondrial matrix-targeted photoactivable GFP that 
            enables detection and quantification of organelle fusion in living 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    mito-PAGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4759)
  AUTHORS   Karbowski M, Arnoult D, Chen H, Chan DC, Smith CL, Youle RJ
  TITLE     Quantitation of mitochondrial dynamics by photolabeling of 
            individual organelles shows that mitochondrial fusion is blocked 
            during the Bax activation phase of apoptosis.
  JOURNAL   J Cell Biol. 2004 Feb 16. 164(4):493-9.
  PUBMED    14769861
REFERENCE   2  (bases 1 to 4759)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 20, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4759
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     196..216
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(241..345)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      372..827
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(459..478)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     669..690
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    834..955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(845..864)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     899..918
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1081..1797)
                     /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(1123..1144)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1472..1491
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1731..1752
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1794..1803
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(1831..1905)
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     promoter        complement(1934..2137)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1963..1983)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2138..2441
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2616..3204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2696..2715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3533..3580
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3605..3624
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3812..4606)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3923..3942)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4533..4552
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4641..239)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4655..31
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4709..4728)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
       61 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
      121 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
      181 ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg
      241 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
      301 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      361 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      421 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      481 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      541 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      601 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      661 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      721 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      781 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttacg ccttaagata
      841 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
      901 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
      961 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     1021 caagtaaaac ctctacaaat gtggtatggc tgattatgat ctagagtcgc ggccgcttta
     1081 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     1141 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactggtggc tcaggtagtg
     1201 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     1261 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttggcct tgatgccgtt
     1321 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
     1381 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     1441 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     1501 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     1561 catgtggtcg gggtagcggc tgaagcactg cacgccgtag ctgaaggtgg tcacgagggt
     1621 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
     1681 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
     1741 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt
     1801 ggcgaccggt ggatccccca acgaatggat cttggcgcgc ggcactggga gccgccgggc
     1861 cgagcctgtc aagccccgca gcagcagcgg cgtcaggacg gacatgctag cggatctgac
     1921 ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     1981 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     2041 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     2101 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     2161 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     2221 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     2281 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     2341 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     2401 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     2461 tatgaactaa tgaccccgta attgattact attaataact aatgcatggc ggtaatacgg
     2521 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2581 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2641 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2701 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2761 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2821 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2881 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2941 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3001 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3061 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3241 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3301 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3361 cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg gccttcaccc
     3421 gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt
     3481 ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac
     3541 gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc
     3601 ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa gaactccagc
     3661 atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc
     3721 aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct
     3781 tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga
     3841 aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc
     3901 attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt
     3961 ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga
     4021 tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg
     4081 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
     4141 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
     4201 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
     4261 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
     4321 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
     4381 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg
     4441 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     4501 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     4561 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc
     4621 atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     4681 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt
     4741 cagccatggg gcggagaat
//
