LOCUS       sequence-247626-        4735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  visualization of mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4735)
  AUTHORS   Friedman JR, Lackner LL, West M, DiBenedetto JR, Nunnari J, Voeltz 
            GK
  TITLE     ER tubules mark sites of mitochondrial division.
  JOURNAL   Science. 2011 Oct 21;334(6054):358-62. doi: 10.1126/science.1207385.
            Epub 2011 Sep 1.
  PUBMED    21885730
REFERENCE   2  (bases 1 to 4735)
  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: 
            "10.1126/science.1207385"; journalName: "Science"; date: 
            "2011-10-21- 21"; volume: "334"; issue: "6054"; pages: "358-62"
COMMENT     Sequence Label: mito-BFP
FEATURES             Location/Qualifiers
     source          1..4735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             673..738
                     /codon_start=1
                     /product="mitochondrial presequence of yeast cytochrome c 
                     oxidase subunit IV (Hurt et al., 1984)"
                     /label=pre-CoxIV
                     /translation="MLSLRQSIRFFKPATRTLCSSR"
     CDS             754..1455
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    1577..1698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1614..1633)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1668..1687
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1705..2160)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1842..1863)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2054..2073
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2187..2291
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2293..2650
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2293..2484
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2316..2336)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2501..2636
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2685..3479
                     /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             2685..3479
                     /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(2739..2758)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3349..3368
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3667..3686)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3711..3758
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4087..4675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4576..4595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
      661 tcagatctcg agatgctttc actacgtcaa tctataagat ttttcaagcc agccacaaga
      721 actttgtgta gctctagacg ggatccaccg gtcatgagcg agctgattaa ggagaacatg
      781 cacatgaagc tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag
      841 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc
      901 cctctcccct tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc
      961 atcaaccaca cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca
     1021 tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc
     1081 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac
     1141 ggccctgtga tgcagaagaa aacactcggc tgggaggcct tcaccgagac gctgtacccc
     1201 gctgacggcg gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat
     1261 ctgatcgcaa acatcaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg
     1321 cctggcgtct actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc
     1381 tacgtcgagc agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg
     1441 cacaagctta attgagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     1501 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     1561 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1621 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1681 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     1741 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     1801 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     1861 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     1921 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     1981 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2041 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2101 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2161 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2221 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2281 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     2341 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2401 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2461 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2521 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2581 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2641 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     2701 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     2761 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     2821 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     2881 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     2941 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3001 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3061 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3121 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3181 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     3241 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3301 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3361 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3421 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3481 cgggactctg gggttcgaaa tgaccgacca agcgatgccc aacctgccat cacgagattt
     3541 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     3601 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     3661 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     3721 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3781 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3841 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3901 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     3961 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4021 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4081 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4141 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4201 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4261 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4321 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4381 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4441 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4501 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4561 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4621 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4681 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatg
//
