LOCUS       sequence-311053-        6389 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  ""Rescue mfn2 expression by adding the guide-resistant-mutant mfn2 
            sequence"".
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6389)
  AUTHORS   Zhou W, Hsu AY, Wang Y, Syahirah R, Wang T, Jeffries J, Wang X, 
            Mohammad H, Seleem MN, Umulis D, Deng Q
  TITLE     Mitofusin 2 regulates neutrophil adhesive migration and the actin 
            cytoskeleton.
  JOURNAL   J Cell Sci. 2020 Sep 4;133(17). pii: jcs.248880. doi: 
            10.1242/jcs.248880.
  PUBMED    32788232
REFERENCE   2  (bases 1 to 6389)
  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."; date: "2020-09-4"; volume: "133(17). pii: jcs.248880. doi"; 
            pages: " 10.1242/jcs.248880"
COMMENT     Sequence Label: plko-shmfn2-mfn2-Rescue
FEATURES             Location/Qualifiers
     source          1..6389
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    172..297
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    790..1023
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1208..1252
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        1550..1798
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1550..1570
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1721..1740
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        1919..2222
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2223..2426
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2377..2397
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     RBS             3388..3396
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     misc_feature    4785..4902
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4951..5461
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     5380..5398
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             5483..6082
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5483..5502)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5979..5999
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
       61 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
      121 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
      181 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
      241 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
      301 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
      361 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
      421 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
      481 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
      541 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
      601 gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg
      661 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
      721 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
      781 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
      841 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
      901 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
      961 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     1021 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     1081 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     1141 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     1201 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     1261 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     1321 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     1381 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     1441 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     1501 tcgacggtat cgatcacgag actagcctcg agcggccgcc cccttcaccg agggcctatt
     1561 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
     1621 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
     1681 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
     1741 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg
     1801 gcaggtttac tgcgaggaaa tctcgagatt tcctcgcagt aaacctgctt tttgaattca
     1861 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     1921 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     1981 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     2041 gggtggagta tttacggtaa gctgcccact tggcagtaca tcaagtgtat catatgccaa
     2101 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     2161 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     2221 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     2281 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     2341 actttccaaa atgtcgtaag aactccgccc cattgacgca aatgggcggt aggcgtgtac
     2401 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctccc
     2461 ggtcgccacc atgtccctgc tcttctctcg atgcaactct atcgtcacag tcaagaaaaa
     2521 taagagacac atggctgagg tgaatgcatc cccacttaag cactttgtca ctgccaagaa
     2581 gaagatcaat ggcatttttg agcagctggg ggcctacatc caggagagcg ccaccttcct
     2641 tgaagacacg tacaggaatg cagaactgga ccccgttacc acagaagaac aggttctgga
     2701 cgtcaaaggt tacctatcca aagtgagagg catcagtgag gtgctggctc ggaggcacat
     2761 gaaagtggct ttttttggcc ggacgagcaa tgggaagagc accgtgatca atgccatgct
     2821 ctgggacaaa gttctgccct ctgggattgg ccacaccacc aattgcttcc tgcgggtaga
     2881 gggcacagat ggccatgagg cctttctcct taccgagggc tcagaggaaa agaggagtgc
     2941 caagactgtg aaccagctgg cccatgccct ccaccaggac aagcagctcc atgccggcag
     3001 cctagtgagt gtgatgtggc ccaactctaa gtgcccactt ctgaaggatg acctcgtttt
     3061 gatggacagc cctggtattg atgtcaccac agagctggac agctggattg acaagttttg
     3121 tctggatgct gatgtgtttg tgctggtggc caactcagag tccaccctga tgcagacgga
     3181 aaagcacttc ttccacaagg tgagtgagcg tctctcccgg ccaaacatct tcatcctgaa
     3241 caaccgctgg gatgcatctg cctcagagcc cgagtacatg gaggaggtgc ggcggcagca
     3301 catggagcgt tgtaccagct tcctggtgga tgagctgggc gtggtggatc gatcccaggc
     3361 cggggaccgc atcttctttg tgtctgctaa ggaggtgctc aacgccagga ttcagaaagc
     3421 ccagggcatg cctgaaggag ggggcgctct cgcagaaggc tttcaagtga ggatgtttga
     3481 gtttcagaat tttgagagga gatttgagga gtgcatctcc cagtctgcag tgaagaccaa
     3541 gtttgagcag cacacggtcc gggccaagca gattgcagag gcggttcgac tcatcatgga
     3601 ctccctgcac atggcggctc gggagcaaca agtgtattgt gaagagatgc gtgaagagcg
     3661 gcaagaccga ctgaaattta ttgacaaaca gctggagctc ttggctcaag actataagct
     3721 gcgaattaag cagattacgg aggaagtgga gaggcaggtg tcgactgcaa tggccgagga
     3781 gatcaggcgc ctctctgtac tggtggacga ttaccagatg gacttccacc cttctccagt
     3841 agtcctcaag gtttataaga atgagctgca ccgccacata gaggaaggac tgggtcgaaa
     3901 catgtctgac cgctgctcca cggccatcac caactccctg cagaccatgc agcaggacat
     3961 gatagatggc ttgaaacccc tccttcctgt gtctgtgcgg agtcagatag acatgctggt
     4021 cccacgccag tgcttctccc tcaactatga cctaaactgt gacaagctgt gtgctgactt
     4081 ccaggaagac attgagttcc atttctctct cggatggacc atgctggtga ataggttcct
     4141 gggccccaag aacagccgtc gggccttgat gggctacaat gaccaggtcc agcgtcccat
     4201 ccctctgacg ccagccaacc ccagcatgcc cccactgcca cagggctcgc tcacccagga
     4261 ggagttcatg gtttccatgg ttaccggcct ggcctccttg acatccagga cctccatggg
     4321 cattcttgtt gttggaggag tggtgtggaa ggcagtgggc tggcggctca ttgccctctc
     4381 ctttgggctc tatggcctcc tctacgtcta tgagcgtctg acctggacca ccaaggccaa
     4441 ggagagggcc ttcaagcgcc agtttgtgga gcatgccagc gagaagctgc agcttgtcat
     4501 cagctacact ggctccaact gcagccacca agtccagcag gaactgtctg ggacctttgc
     4561 tcatctgtgt cagcaagttg acgtcacccg ggagaacctg gagcaggaaa ttgccgccat
     4621 gaacaagaaa attgaggttc ttgactcact tcagagcaaa gcaaagctgc tcaggaataa
     4681 agccggttgg ttggacagtg agctcaacat gttcacacac cagtacctgc agcccagcag
     4741 ataaaattct cgacctcgag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     4801 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     4861 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     4921 agcagagatc cactttggcc gcggctcgag ggggttgggg ttgcgccttt tccaaggcag
     4981 ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc
     5041 cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc
     5101 tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt
     5161 gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc
     5221 agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag
     5281 cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg
     5341 gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc
     5401 tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca
     5461 gggggatcca ccggagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     5521 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     5581 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     5641 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     5701 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     5761 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     5821 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     5881 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     5941 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     6001 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     6061 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     6121 cgcacgaccc catgcatcgg tacctttaag accaatgact tacaaggcag ctgtagatct
     6181 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     6241 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     6301 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     6361 agtagtgtgt gcccgtctgt tgtgtgact
//
