LOCUS       sequence-379817-        6526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A pMSCV-based backbone where the MCS has been substituted with a 
            MCS-IRES-mitoYFP for expression of a gene of interest and a 
            mitochondrially targeted YFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6526)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6526)
  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            "unpublished"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: pMIMY
FEATURES             Location/Qualifiers
     source          1..6526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(45..461)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(61..77)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(93..115)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    528..869
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             933..1449
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(1649..1668)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1796..1818)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2012..2031)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2131..2153
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2191..2209)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2277..2381
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2382..2450,2451..3242)
                     /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(2600..2619)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3413..4001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3902..3921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4155..4172
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4289..4310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4325..4330,4331..4348,4349..4355)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4363..4379
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     LTR             4541..5055
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(5122..5832,5833..5835,5836..5838))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERAIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5164..5185)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5772..5793
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(5872..5946)
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     misc_feature    5962..6514
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6124..6143)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     6353..6370
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 gnsskkgsks rscrkrrttc gttaacctcg agagatctaa ttccggcgcc tagagaagga
       61 gtgagggctg gataaaggga ggatcgaggc ggggtcgaac gaggaggttc aagggggaga
      121 gacggggcgg atggaggaag aggaggcgga ggcttagggt gtacaaaggg cttgacccag
      181 ggaggggggt caaaagccaa ggcttcccag gtcacgatgt aggggacctg gtctgggtgt
      241 ccatgcgggc caggtgaaaa gaccttgatc ttaacctggg tgatgaggtc tcggttaaag
      301 gtgccgtctc gcggccatcc gacgttaaag gttggccatt ctgcagagca gaaggtaacc
      361 caacgtctct tcttgacatc taccgactgg ttgtgagcga tccgctcgac atctttccag
      421 tgacctaagg tcaaacttaa gggagtggta acagtctggc cctaattttc agacaaatac
      481 agaaacacag tcagacagag acaacacaga acgatgctgc agcgctgcag cagacaagac
      541 gcgcggcttc ggttccaaac cgaaagcaaa aattcagacg gaggcgggaa ctgttttagg
      601 ttctcgtctc ctaccagaac cacatatcct gacggggtcg gattccacat cgactccctt
      661 cctcaggtcg ggccacaaaa acggccccca aagtccctgg gacgtctccc agggttgcgg
      721 ccgggtgttc agaactcgtc agttccacca cgggtccgcc agatacagag ctagttagct
      781 aactagtaca gacgcaggcg caaacattag atgccggcac aaacacgcac aaagacagag
      841 acagacacga aacgaccgct ggccagctta cctcccggcg ggggggtcgg tggtccctgg
      901 gcaggggtct ccaaatctcg gtggaacctc caaatgaaag acccccgagg tgggcagtca
      961 atcaatctga ggagaccctc ccaaggatca gcgagtccac gattcggatg caaacagcaa
     1021 gaggctttat tgggaatacg ggtacccggg cgacgcagtc tatcggagga ctggcgcgcc
     1081 gagtgagggg ttgtgggctc ttttattgag ctcggggagc agaagcgcgc gaacagaagc
     1141 gagaagcgaa ctgattggtt agttcaaata aggcacaggg tcatttcagg tccttggggc
     1201 accctggaaa catctgatgg ttctctagaa actgctgagg gcgggaccgc atctggggac
     1261 catctgttct tggccctgag ccggggcagg aactgcttac cacagatatc ctgtttggcc
     1321 catattctgc tgtctctctg ttcctaacct tgatctgaac ttctctattc tcagttatgt
     1381 attttccatg ccttgcaaaa tggcgttact taagctagct tgccaaacct acaggtgggg
     1441 tctttcattc cccccttttt ctggagacta aataaaatct tttattttat ctatggctcg
     1501 tactctatag gcttcagctg gtgatattgt tgagtcaaaa ctagagcctg gaccactgat
     1561 atcctgtctt taacaaattg gactaatcgc ctctacgccg gacgcatcgt ggccggcatc
     1621 accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat
     1681 cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc
     1741 gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc gccgtcgttt
     1801 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     1861 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     1921 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     1981 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     2041 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     2101 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     2161 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     2221 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2281 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2341 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2401 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2461 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2521 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2581 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2641 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2701 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2761 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2821 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2881 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2941 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3001 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3061 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3121 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3181 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3241 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3301 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3361 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3421 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3481 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3541 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3601 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3661 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3721 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3781 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3841 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3901 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3961 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4021 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4081 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4141 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     4201 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     4261 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     4321 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     4381 ttcacaattc tccccagctc tttctgcgct gtccataatg aactatttca ggagtattgg
     4441 agtatgtgga ttcaaaacag tggctaattg agtcaggatt acttaatatt cgactcctac
     4501 cctcagaatg ttgttctcca acttcaagaa actgttaccc atgaaagacc cccgctgacg
     4561 ggtagtcaat cactcagagg agaccctccc aaggaacagc gagaccacaa gtcggatgca
     4621 actgcaagag ggtttattgg atacacgggt acccgggcga cgcagtctat cggaggactg
     4681 gcgcgccgag tgaggggttg tgggctcttt tattgagctc ggggagcaga agcgcgcgaa
     4741 cagaagcgag aagcgaactg attggttagt tcaaataagg cacagggtca tttcaggtcc
     4801 ttggggcacc ctggaaacat ctgatggttc tctagaaact gctgagggcg ggaccgcatc
     4861 tggggaccat ctgttcttgg ccctgagccg gggcaggaac tgcttaccac agatatcctg
     4921 tttggcccat attctgctgt ctctctgttc ctaaccttga tctgaacttc tctattctca
     4981 gttatgtatt ttccatgcct tgcaaaatgg cgttacttaa gctagcttgc caaacctaca
     5041 ggtggggtct ttcattcccc cctttttctg gagactaaat aaaatctttt attttatcga
     5101 taagcttggc tgcaggtcga ccttgtacag ctcgtccatg ccgagagtga tcccggcggc
     5161 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc
     5221 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     5281 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
     5341 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
     5401 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     5461 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttata
     5521 gttgccgtcg tccttgaaga agatggcgcg ctcctggacg tagccttcgg gcatggcgga
     5581 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcaggccgta
     5641 gccgaaggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
     5701 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     5761 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     5821 ctcgcccttg ctcaccatcg tggcgaccgg tggatccccc aacgaatgga tcttggcgcg
     5881 cggcactggg agccgccggg ccgagcctgt caagccccgc agcagcagcg gcgtcaggac
     5941 ggacatccat ggttgtggcc atattatcat cgtgtttttc aaaggaaaac cacgtccccg
     6001 tggttcgggg ggcctagacg tttttttaac ctcgactaaa cacatgtaaa gcatgtgcac
     6061 cgaggcccca gatcagatcc catacaatgg ggtaccttct gggcatcctt cagccccttg
     6121 ttgaatacgc ttgaggagag ccatttgact ctttccacaa ctatccaact cacaacgtgg
     6181 cactggggtt gtgccgcctt tgcaggtgta tcttatacac gtggcttttg gccgcagagg
     6241 cacctgtcgc caggtggggg gttccgctgc ctgcaaaggg tcgctacaga cgttgtttgt
     6301 cttcaagaag cttccagagg aactgcttcc ttcacgacat tcaacagacc ttgcattcct
     6361 ttggcgagag gggaaagacc cctaggaatg ctcgtcaaga agacagggcc aggtttccgg
     6421 gccctcacat tgccaaaaga cggcaatatg gtggaaaata acatatagac aaacgcacac
     6481 cggccttatt ccaagcggct tcggccagta acgttagggg gggggg
//
