LOCUS       sequence-342370-        5073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Mitochondria, Excitation: 448 / 473, Emission: 500 / 
            528.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5073)
  AUTHORS   Shu X, Lev-Ram V, Deerinck TJ, Qi Y, Ramko EB, Davidson MW, Jin Y, 
            Ellisman MH, Tsien RY
  TITLE     A genetically encoded tag for correlated light and electron 
            microscopy of intact cells, tissues, and organisms.
  JOURNAL   PLoS Biol. 2011 Apr;9(4):e1001041. doi: 
            10.1371/journal.pbio.1001041. Epub 2011 Apr 5.
  PUBMED    21483721
REFERENCE   2  (bases 1 to 5073)
  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: "PLoS 
            Biol."; date: "2011-04"; volume: "9(4):e1001041. doi"; pages: " 
            10.1371/journal.pbio.1001041. Epub 2011 Apr 5"
COMMENT     Sequence Label: PA-mCherry-miniSOG-Mito-7
FEATURES             Location/Qualifiers
     source          1..5073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             538..612
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      640..649
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             646..1353
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein (Subach et al., 2009)"
                     /label=PAmCherry1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
                     GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(791..809)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             1360..1680
                     /codon_start=1
                     /product="mini singlet oxygen generator, a fluorescent 
                     flavoprotein suitable for correlated light and electron 
                     microscopy (Shu et al., 2011)"
                     /label=miniSOG
                     /translation="MEKSFVITDPRLPDNPIIFASDGFLELTEYSREEILGRNGRFLQG
                     PETDQATVQKIRDAIRDQREITVQLINYTKSGKKFWNLLHLQPMRDQKGELQYFIGVQL
                     DG"
     polyA_signal    1802..1923
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1839..1858)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1893..1912
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1930..2385)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2067..2088)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2279..2298
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2412..2516
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2518..2875
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2518..2709
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2541..2561)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2726..2861
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2910..3704
                     /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             2910..3704
                     /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(2964..2983)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3574..3593
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3892..3911)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3936..3983
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4312..4900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4801..4820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcatg
      541 tccgtcctga cgccgctgct gctgcggggc ttgacaggct cggcccggcg gctcccagtg
      601 ccgcgcgcca agatccattc gttgggggat ccaccggtcg ccaccatggt gagcaagggc
      661 gaggaggata acatggccat cattaaggag ttcatgcgct tcaaggtgca catggagggg
      721 tccgtgaacg gccacgtgtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
      781 acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcac ctgggacatc
      841 ctgagccctc agttcatgta cggctccaat gcctacgtga agcaccccgc cgacatcccc
      901 gactacttta agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaaattcgag
      961 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     1021 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtgatgca gaagaagacc
     1081 atgggctggg aggccctctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     1141 gtcaagccca gagtgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     1201 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaaccg caagctggac
     1261 atcaccagcc acaacgagga ctacaccatc gtggagcagt acgagagagc cgagggccgc
     1321 cactccaccg gcggcatgga cgagctgtac aagggcggca tggagaagag cttcgtgatc
     1381 accgaccccc gcctgcccga caaccccatc atcttcgcca gcgacggctt cctggagctg
     1441 accgagtaca gcagagagga gatcctgggc cgcaacggcc gcttcctgca gggccccgag
     1501 accgaccagg ccaccgtgca gaagatccgc gacgccatcc gcgaccagcg cgagatcacc
     1561 gtgcagctga tcaactacac caagagcggc aagaagttct ggaacctgct gcacctgcag
     1621 cccatgcgcg accagaaggg cgagctgcag tacttcatcg gcgtgcagct ggacggctaa
     1681 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     1741 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     1801 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1861 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1921 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     1981 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2041 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2101 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2161 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2221 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2281 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2341 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     2401 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2461 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     2521 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2581 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2641 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2701 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2761 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     2821 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     2881 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     2941 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3001 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3061 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     3121 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3181 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3241 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3301 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3361 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3421 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3481 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3541 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3601 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3661 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     3721 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     3781 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3841 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     3901 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     3961 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4021 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4081 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4141 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     4201 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4261 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4321 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4381 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4441 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4501 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4561 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4621 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4681 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4741 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4801 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4861 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4921 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4981 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
     5041 ggtcattagt tcatagccca tatatggagt tcc
//
