LOCUS       sequence-379552-        6453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Exogenous expression of BAK-YFP chimera in mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6453)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6453)
  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            "unpublished"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: pMSCV-YFP-Bak-mito
FEATURES             Location/Qualifiers
     source          1..6453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(760..1551,1552..1620))
                     /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     1383..1402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1621..1725)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1793..1811
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1849..1871)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1971..1990
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2184..2206
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2334..2353
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2553..3069
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    3133..3474
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3541..3957
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3887..3909
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3925..3941
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      3982..3991
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3988..3990,3991..3993,3994..4704)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4033..4054)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4294..4313)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4641..4662
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5400..5914
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6076..6092
                     /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)."
     promoter        complement(join(6100..6106,6107..6124,6125..6130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6145..6166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6283..6300)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1741 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     1801 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1861 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1921 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     1981 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     2041 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     2101 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     2161 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat
     2221 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     2281 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     2341 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     2401 ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta
     2461 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
     2521 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     2581 aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga
     2641 agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt
     2701 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg
     2761 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     2821 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     2881 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
     2941 agactgcgtc gcccgggtac ccgtattccc aataaagcct cttgctgttt gcatccgaat
     3001 cgtggactcg ctgatccttg ggagggtctc ctcagattga ttgactgccc acctcggggg
     3061 tctttcattt ggaggttcca ccgagatttg gagacccctg cccagggacc accgaccccc
     3121 ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt
     3181 tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt tagctaacta gctctgtatc
     3241 tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt
     3301 cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat
     3361 ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct
     3421 ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc
     3481 tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag
     3541 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
     3601 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
     3661 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
     3721 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
     3781 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
     3841 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
     3901 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga
     3961 attcgttaac ctcgaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     4021 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     4081 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     4141 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgcagtgc
     4201 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     4261 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     4321 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     4381 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     4441 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     4501 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     4561 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac
     4621 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     4681 ctcggcatgg acgagctgta caagtccgga ctcagatctg catctggaca aggaccaggt
     4741 cccccgaagg tgggctgcga tgagtccccg tccccttctg aacagcaggt tgcccaggac
     4801 acagaggagg tctttcgaag ctacgttttt tacctccacc agcaggaaca ggagacccag
     4861 ggggcggccg cccctgccaa ccccgagatg gacaacttgc ccctggaacc caacagcatc
     4921 ttgggtcagg tgggtcggca gcttgctctc atcggagatg atattaaccg gcgctacgac
     4981 acagagttcc agaatttact agaacagctt cagcccacag ccgggaatgc ctacgaactc
     5041 ttcaccaaga tcgcctccag cctatttaag agtggcatca gctggggccg cgtggtggct
     5101 ctcctgggct ttggctaccg tctggccctg tacgtctacc agcgtggttt gaccggcttc
     5161 ctgggccagg tgacctgctt tttggctgat atcatactgc atcattacat cgccagatgg
     5221 atcgcacaga gaggcggttg ggtggcagcc ctgaatttgc gtagagactt aattcttgca
     5281 atgttagcta ttggcgtgtt ctctttaggg gcgtttatca aaattattca attaagaaaa
     5341 aataattaag cttatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     5401 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     5461 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
     5521 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     5581 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
     5641 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     5701 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     5761 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata
     5821 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     5881 agtgattgac tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga
     5941 acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt
     6001 tgaatccaca tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg
     6061 gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6121 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6181 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6241 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6301 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6361 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6421 caggaaccgt aaaaaggccg cgttgctggc gtt
//
