LOCUS       sequence-472857-        6498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Elutable MitoTag-AP system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6498)
  AUTHORS   Cunningham CN, Van Vranken JG, Larios J, Heyden K, Gygi SP, Rutter J
  TITLE     A dual-purification system to isolate mitochondrial subpopulations.
  JOURNAL   J Cell Sci. 2025 Mar 13:jcs.263693. doi: 10.1242/jcs.263693.
  PUBMED    40079232
REFERENCE   2  (bases 1 to 6498)
  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: "J Cell Sci.
            2025 Mar 13:jcs.263693. doi: 10.1242/jcs.263693."
COMMENT     Sequence Label: pMXs-8XHis-mCherry-Omp25
FEATURES             Location/Qualifiers
     source          1..6498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             890..1359
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(1530..1925)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    join(1926..1934,1935..1937,1938..2512)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2099..2118)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2328..2345
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(2663..3367)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2938..2957)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3207..3225
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(3380..3403)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     misc_feature    3468..3842
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     complement(3514..3530)
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             complement(3852..4268)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3868..3884)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3900..3922)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4327..4684
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             4747..5340
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        5370..5474
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5475..5543,5544..6335)
                     /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     complement(5693..5712)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaactatg gccatttaat
      841 gtaaatactt aagaaaaaaa accaaattaa ttttgataca tgctgcatgt gaaagacccc
      901 cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga gaccacaagt
      961 cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgact cagtcaatcg
     1021 gaggactggc gccccgagtg aggggttgtg ggctctttta ttgagctcgg ggagcagaag
     1081 cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca cagggtcatt
     1141 tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc tgagggctgg
     1201 accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg cttaccacag
     1261 atatcctgtt tggcccatat tcagctgttc catctgttct tggccctgag ccggggcagg
     1321 aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct gttcctgacc
     1381 ttgatctgaa cttctctatt ctcagttatg tatttttcca tgccttgcaa aatggcgtta
     1441 cttaagctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt ttctggagac
     1501 taaataaaat cttttatttt atcgtcgact tagccctccc acacataacc agagggcagc
     1561 aattcacgaa tcccaactgc cgtcggctgt ccatcactgt ccttcactat ggctttgatc
     1621 ccaggatgca gatcgagaag cacctgtcgg caccgtccgc aggggctcaa gatgcccctg
     1681 ttctcatttc cgatcgcgac gatacaagtc aggttgccag ctgccgcagc agcagcagtg
     1741 cccagcacca cgagttctgc acaaggtccc ccagtaaaat gatatacatt gacaccagtg
     1801 aagatgcggc cgtcgctaga gagagctgcg ctggcgacgc tgtagtcttc agagatgggg
     1861 atgctgttga ttgtagccgt tgctctttca atgagggtgg attcttcttg agacaaagga
     1921 accatggttg tggccatatt atcatcgtgt ttttcaaagg aaaaccacgt ccccgtggtt
     1981 cggggggcct agacgttttt ttaacctcga ctaaacacat gtaaagcatg tgcaccgagg
     2041 ccccagatca gatcccatac aatggggtac cttctgggca tccttcagcc ccttgttgaa
     2101 tacgcttgag gagagccatt tgactctttc cacaactatc caactcacaa cgtggcactg
     2161 gggttgtgcc gcctttgcag gtgtatctta tacacgtggc ttttggccgc agaggcacct
     2221 gtcgccaggt ggggggttcc gctgcctgca aagggtcgct acagacgttg tttgtcttca
     2281 agaagcttcc agaggaactg cttccttcac gacattcaac agaccttgca ttcctttggc
     2341 gagaggggaa agacccctag gaatgctcgt caagaagaca gggccaggtt tccgggccct
     2401 cacattgcca aaagacggca atatggtgga aaataacata tagacaaacg cacaccggcc
     2461 ttattccaag cggcttcggc cagtaacgtt aggggggggg gagggagagg gggggggggg
     2521 gcggaattta cgtagcggcc gctcagagct gctttcggta tctcacgaag gcccaaactg
     2581 ctaccagggt aagggcaaac actggcagca gcaccacagc tacaggaact ccactaggct
     2641 ctccgtcgcc tcgatgccgc ggcttgtaca gctcgtccat gccgccggtg gagtggcggc
     2701 cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca
     2761 acttgatgtt gacgttgtag gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg
     2821 tcttgacctc agcgtcgtag tggccgccgt ccttcagctt cagcctctgc ttgatctcgc
     2881 ccttcagggc gccgtcctcg gggtacatcc gctcggagga ggcctcccag cccatggtct
     2941 tcttctgcat tacggggccg tcggagggga agttggtgcc gcgcagcttc accttgtaga
     3001 tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt caccacgccg ccgtcctcga
     3061 agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcaag tagtcgggga
     3121 tgtcggcggg gtgcttcacg taggccttgg agccgtacat gaactgaggg gacaggatgt
     3181 cccaggcgaa gggcaggggg ccacccttgg tcaccttcag cttggcggtc tgggtgccct
     3241 cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc acggagccct
     3301 ccatgtgcac cttgaagcgc atgaactcct tgatgatggc catgttatcc tcctcgccct
     3361 tgctcacgcc gctccctccg tgatggtgat ggtgatggtg atgctcgagg cctgcaggaa
     3421 ttcccgtacc accacactgg gatccttaat taactagcta gatccggcag tctagaggat
     3481 ggtccacccc cggggtcggc agccttcacg tgggcggcgt gtatccaagc tgcgatgccg
     3541 tctactttga gggcggtggg ggtggtcagc aggactgtgt aaggtccttt ccagcgaggt
     3601 tctaggttct tagtctggtg tcggcggacc cacactgtgt cgccgactcg gtaagggtga
     3661 ggtaccaccg gtcggtccag ttgttcttgg taggctgccg ccagaggtct ccagacttcg
     3721 tgctggacta agtagagagc ctgtaagtga gcttggagag aggggctgtt agtaactctt
     3781 gtcatgtcag ggtcagggaa gtttacaagg ggcgggggtg ccccatataa gatctcatat
     3841 ggccatatgg gggcgcctag agaaggagtg agggctggat aaagggagga tcgaggcggg
     3901 gtcgaacgag gaggttcaag ggggagagac ggggcggatg gaggaagagg aggcggaggc
     3961 ttagggtgta caaagggctt gacccaggga ggggggtcaa aagccaaggc ttcccaggtc
     4021 acgatgtagg ggacctggtc tgggtgtcca tgcgggccag gtgaaaagac cttgatctta
     4081 acctgggtga tgaggtctcg gttaaaggtg ccgtctcgcg gccatccgac gttaaaggtt
     4141 ggccattctg cagagcagaa ggtaacccaa cgtctcttct tgacatctac cgactggttg
     4201 tgagcgatcc gctcgacatc tttccagtga cctaaggtca aacttaaggg agtggtaaca
     4261 gtctggccct aattttcaga caaatacaga aacacagtca gacagagaca acacagaacg
     4321 atgctgcagc agacaagacg cgcggcgcgg cttcggtccc aaaccgaaag caaaaattca
     4381 gacggaggcg ggaactgttt taggttctcg tctcctacca gaaccacata tccctcctat
     4441 taggggggtg caccaaagag tccaaaacga tcgggatttt tggactcagg tcgggccaca
     4501 aaaacggccc ccgaagtccc tgggacgtct cccagggttg cggccgggtg ttccgaactc
     4561 gtcagttcca ccacgggtcc gccagataca gagctagtta gctaactagt accgacgcag
     4621 gcgcataaaa tcagtcatag acactagaca atcggacaga cacagataag ttgctggcca
     4681 gcttacctcc cggtggtggg tcggtggtcc ctgggcaggg gtctcccgat cccggacgag
     4741 cccccaaatg aaagaccccc gctgacgggt agtcaatcac tcagaggaga ccctcccaag
     4801 gaacagcgag accacaagtc ggatgcaact gcaagagggt ttattggata cacgggtacc
     4861 cgggcgactc agtcaatcgg aggactggcg cgccgagtga ggggttgtgg gctcttttat
     4921 tgagctcggg gagcagaagc gcgcgaacag aagcgagaag cgaactgatt ggttagttca
     4981 aataaggcac agggtcattt caggtccttg gggcaccctg gaaacatctg atggttctct
     5041 agaaactgct gagggctgga ccgcatctgg ggaccatctg ttcttggccc tgagccgggg
     5101 caggaactgc ttaccacaga tatcctgttt ggcccatatt cagctgttcc atctgttctt
     5161 ggccctgagc cggggcagga actgcttacc acagatatcc tgtttggccc atattcagct
     5221 gttccatctg ttcctgacct tgatctgaac ttttctattc tcagttatgt atttttccat
     5281 gccttgcaaa atggcgttac ttaagctagc ttgccaaacc tacaggtggg gtctttcatt
     5341 atgcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5401 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5461 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5521 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5581 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5641 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5701 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5761 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5821 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5881 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5941 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6001 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6061 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6121 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6181 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6241 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6301 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6361 tactttagat tgatttgcgg ccggccgcaa aaggatctag gtgaagatcc tttttgataa
     6421 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6481 aaagatcaaa ggatcttc
//
