LOCUS       sequence-474211-        4802 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses photoactivable fluorescent reporter for visualizing 
            mitochondrial network.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4802)
  AUTHORS   Mondal, P
  TITLE     Photoactivable plasmid labeling Mitochondria for Mammalian 
            expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4802)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Mito-Dendra2
FEATURES             Location/Qualifiers
     source          1..4802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(76..97)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     288..307
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        421..525
                     /gene="bla"
                     /label=AmpR promoter
     promoter        527..884
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        527..718
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(550..570)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      735..870
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             919..1713
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             919..1713
                     /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(973..992)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1583..1602
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1901..1920)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1945..1992
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2321..2909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2810..2829
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3084..3387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3388..3591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3542..3562
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3620..3694
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             3719..4408
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     polyA_signal    4613..4734
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4650..4669)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4704..4723
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(4741..4802,1..394))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
       61 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
      121 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
      181 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
      241 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
      301 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
      361 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
      421 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      481 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
      541 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
      601 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
      661 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
      721 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
      781 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
      841 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
      901 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
      961 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     1021 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     1081 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     1141 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     1201 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     1261 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     1321 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     1381 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     1441 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     1501 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     1561 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     1621 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     1681 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     1741 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     1801 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1861 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     1921 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     1981 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     2041 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     2101 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     2161 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2221 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2281 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2341 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2401 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2461 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2521 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2581 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2641 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2701 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2761 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2821 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2881 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2941 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3001 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     3061 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     3121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     3181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     3241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     3301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     3361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     3421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     3481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     3541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     3601 aaccgtcaga tccgctagca tgtccgtcct gacgccgctg ctgctgcggg gcttgacagg
     3661 ctcggcccgg cggctcccag tgccgcgcgc caagatccat tcgttggggg atccaaccat
     3721 gaacaccccg ggaattaacc tgatcaagga ggacatgcgc gtgaaggtgc acatggaggg
     3781 caacgtgaac ggccacgcct tcgtgatcga gggcgagggc aagggcaagc cctacgaggg
     3841 cacccagacc gccaacctga ccgtgaagga gggcgccccc ctgcccttca gctacgacat
     3901 cctgaccacc gccgtgcact acggcaaccg ggtgttcacc aagtaccccg aggacatccc
     3961 cgactacttc aagcagagct tccccgaggg ctacagctgg gagcgcacca tgaccttcga
     4021 ggacaagggc atctgcacca tccgcagcga catcagcctg gagggcgact gcttcttcca
     4081 gaacgtgcgc ttcaagggca ccaacttccc ccccaacggc cccgtgatgc agaagaagac
     4141 cctgaagtgg gagcccagca ccgagaagct gcacgtgcgc gacggcctgc tggtgggcaa
     4201 catcaacatg gccctgctgc tggagggcgg cggccactac ctgtgcgact tcaagaccac
     4261 ctacaaggcc aagaaggtgg tgcagctgcc cgacgcccac ttcgtggacc accgcatcga
     4321 gatcctgggc aacgacagcg actacaacaa ggtgaagctg tacgagcacg ccgtggcccg
     4381 ctacagcccc ctgcccagcc aggtgtggtc cggcgacagc ggcgtgtaca agactcgagt
     4441 tcgtgaccgc cgccgccatc acccacggca tggatgagct gtacaagtaa agcggccgcg
     4501 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     4561 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     4621 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4681 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     4741 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4801 tt
//
