LOCUS       sequence-429090-        6597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For tagging mitochondria with HA epitopes (HA-MITO construct).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6597)
  AUTHORS   Chen WW, Freinkman E, Wang T, Birsoy K, Sabatini DM
  TITLE     Absolute Quantification of Matrix Metabolites Reveals the Dynamics 
            of Mitochondrial Metabolism.
  JOURNAL   Cell. 2016 Aug 25;166(5):1324-1337.e11. doi: 
            10.1016/j.cell.2016.07.040.
  PUBMED    27565352
REFERENCE   2  (bases 1 to 6597)
  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: 
            "10.1016/j.cell.2016.07"; journalName: "Cell"; date: "2016-08-25- 
            25"; volume: "166"; issue: "5"; pages: "1324-1337.e11"
COMMENT     Sequence Label: pMXs-3XHA-EGFP-OMP25
FEATURES             Location/Qualifiers
     source          1..6597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(2..106)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             136..729
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    792..1149
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1208..1624
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1554..1576
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1592..1608
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1634..2008
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1946..1962
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2082..2108
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2121..2147
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2160..2186
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2199..2201,2202..2912)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2199..2201,2202..2912)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2241..2262)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2502..2521)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2849..2870
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(3063..3637,3638..3640,3641..3649)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3230..3247)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3457..3476
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3650..4045
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4216..4685
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4815..4832)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4986..5574)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5066..5085)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5738..6529,6530..6597,1))
                     /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     6361..6380
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
       61 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
      121 aagtgccacc tgcataatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
      181 gccattttgc aaggcatgga aaaatacata actgagaata gaaaagttca gatcaaggtc
      241 aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
      301 ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt
      361 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag
      421 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
      481 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
      541 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
      601 tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
      661 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
      721 tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac
      781 caccgggagg taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg
      841 actgatttta tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc
      901 cgtggtggaa ctgacgagtt cggaacaccc ggccgcaacc ctgggagacg tcccagggac
      961 ttcgggggcc gtttttgtgg cccgacctga gtccaaaaat cccgatcgtt ttggactctt
     1021 tggtgcaccc ccctaatagg agggatatgt ggttctggta ggagacgaga acctaaaaca
     1081 gttcccgcct ccgtctgaat ttttgctttc ggtttgggac cgaagccgcg ccgcgcgtct
     1141 tgtctgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga
     1201 aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt
     1261 cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg
     1321 ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga
     1381 cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca
     1441 ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc
     1501 ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga
     1561 acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg
     1621 cgcccccata tggccatatg agatcttata tggggcaccc ccgccccttg taaacttccc
     1681 tgaccctgac atgacaagag ttactaacag cccctctctc caagctcact tacaggctct
     1741 ctacttagtc cagcacgaag tctggagacc tctggcggca gcctaccaag aacaactgga
     1801 ccgaccggtg gtacctcacc cttaccgagt cggcgacaca gtgtgggtcc gccgacacca
     1861 gactaagaac ctagaacctc gctggaaagg accttacaca gtcctgctga ccacccccac
     1921 cgccctcaaa gtagacggca tcgcagcttg gatacacgcc gcccacgtga aggctgccga
     1981 ccccgggggt ggaccatcct ctagactgcc ggatctagct agttaattaa ggatcccagt
     2041 gtggtggtac gggaattcct gcaggcctcg aggccaccat gtatccctat gacgtgcctg
     2101 attacgccgg cacaggatcc tacccctatg atgtgcctga ctacgctggc agcgccggat
     2161 acccttatga tgtgcctgat tatgctggag ggagcggcgt gagcaagggc gaggagctgt
     2221 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2281 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2341 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     2401 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2461 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2521 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2581 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2641 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     2701 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2761 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     2821 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2881 ggatcactct cggcatggac gagctgtaca agccgcggca tcgaggcgac ggagagccta
     2941 gtggagttcc tgtagctgtg gtgctgctgc cagtgtttgc ccttaccctg gtagcagttt
     3001 gggccttcgt gagataccga aagcagctct gagcggccgc tacgtaaatt ccgccccccc
     3061 cccccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     3121 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     3181 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     3241 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     3301 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     3361 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     3421 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     3481 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     3541 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     3601 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca tggttccttt
     3661 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
     3721 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
     3781 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
     3841 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
     3901 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     3961 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     4021 ctctggttat gtgtgggagg gctaagtcga cgataaaata aaagatttta tttagtctcc
     4081 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     4141 cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag
     4201 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     4261 ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     4321 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc
     4381 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     4441 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     4501 ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg
     4561 actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
     4621 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
     4681 tttcacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat ttacattaaa
     4741 tggccatagt tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4801 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4861 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4921 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4981 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5041 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5101 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5161 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5221 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5281 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5341 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5401 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5461 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5521 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5581 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5641 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttgcggcc ggccgcaaat
     5701 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5761 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5821 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5881 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5941 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6001 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6061 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6121 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6181 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6241 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6301 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6361 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6421 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6481 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6541 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactca
//
