LOCUS       sequence-238029-        8530 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of MFN2 and mitoRFP in mammalial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8530)
  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 8530)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pMYs-MFN2-IRES-mitoRFP
FEATURES             Location/Qualifiers
     source          1..8530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             1103..1519
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1487..1503
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1529..1903
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1841..1857
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     RBS             2915..2923
                     /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    join(4301..4874,4875..4877,4878..4886)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4467..4484)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4694..4713
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4893..4967
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      4995..5004
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5001..5732
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTETLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     primer_bind     complement(6634..6651)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6805..7393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6885..6904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7557..8348,8349..8417))
                     /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     8180..8199
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8418..8522)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat
      541 ccgaatcgtg gactcgctga tccttgggag ggtctcctca gattgattga ctgcccacct
      601 cgggggtctt tcatttggag gttccaccga gatttggaga cccctgccca gggaccaccg
      661 acccccccgc cgggaggtaa gctggccagc ggtcgtttcg tgtctgtctc tgtctttgtg
      721 cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt actagttagc taactagctc
      781 tgtatctggc ggacccgtgg tggaactgac gagttcggaa cacccggccg caaccctggg
      841 agacgtccca gggacttcgg gggccgtttt tgtggcccga cctgagtcca aaaatcccga
      901 tcgttttgga ctctttggtg caccccccta ataggaggga tatgtggttc tggtaggaga
      961 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt gggaccgaag
     1021 ccgcgccgcg cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
     1081 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
     1141 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     1201 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     1261 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     1321 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
     1381 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
     1441 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
     1501 cactccttct ctaggcgccc ccatatggcc atatgagatc ttatatgggg cacccccgcc
     1561 ccttgtaaac ttccctgacc ctgacatgac aagagttact aacagcccct ctctccaagc
     1621 tcacttacag gctctctact tagtccagca cgaagtctgg agacctctgg cggcagccta
     1681 ccaagaacaa ctggaccgac cggtggtacc tcacccttac cgagtcggcg acacagtgtg
     1741 ggtccgccga caccagacta agaacctaga acctcgctgg aaaggacctt acacagtcct
     1801 gctgaccacc cccaccgccc tcaaagtaga cggcatcgca gcttggatac acgccgccca
     1861 cgtgaaggct gccgaccccg ggggtggacc atcctctaga ctgccggatc tagctagtta
     1921 attaaggatc ccagtgtggt ggtacgggaa ttctgcagtc gacggtaccg cgggcccggg
     1981 atccaccggt cgccaccatg tccctgctct tctctcgatg caactctatc gtcacagtca
     2041 agaaaaataa gagacacatg gctgaggtga atgcatcccc acttaagcac tttgtcactg
     2101 ccaagaagaa gatcaatggc atttttgagc agctgggggc ctacatccag gagagcgcca
     2161 ccttccttga agacacgtac aggaatgcag aactggaccc cgttaccaca gaagaacagg
     2221 ttctggacgt caaaggttac ctatccaaag tgagaggcat cagtgaggtg ctggctcgga
     2281 ggcacatgaa agtggctttt tttggccgga cgagcaatgg gaagagcacc gtgatcaatg
     2341 ccatgctctg ggacaaagtt ctgccctctg ggattggcca caccaccaat tgcttcctgc
     2401 gggtagaggg cacagatggc catgaggcct ttctccttac cgagggctca gaggaaaaga
     2461 ggagtgccaa gactgtgaac cagctggccc atgccctcca ccaggacaag cagctccatg
     2521 ccggcagcct agtgagtgtg atgtggccca actctaagtg cccacttctg aaggatgacc
     2581 tcgttttgat ggacagccct ggtattgatg tcaccacaga gctggacagc tggattgaca
     2641 agttttgtct ggatgctgat gtgtttgtgc tggtggccaa ctcagagtcc accctgatgc
     2701 agacggaaaa gcacttcttc cacaaggtga gtgagcgtct ctcccggcca aacatcttca
     2761 tcctgaacaa ccgctgggat gcatctgcct cagagcccga gtacatggag gaggtgcggc
     2821 ggcagcacat ggagcgttgt accagcttcc tggtggatga gctgggcgtg gtggatcgat
     2881 cccaggccgg ggaccgcatc ttctttgtgt ctgctaagga ggtgctcaac gccaggattc
     2941 agaaagccca gggcatgcct gaaggagggg gcgctctcgc agaaggcttt caagtgagga
     3001 tgtttgagtt tcagaatttt gagaggagat ttgaggagtg catctcccag tctgcagtga
     3061 agaccaagtt tgagcagcac acggtccggg ccaagcagat tgcagaggcg gttcgactca
     3121 tcatggactc cctgcacatg gcggctcggg agcaacaagt gtattgtgaa gagatgcgtg
     3181 aagagcggca agaccgactg aaatttattg acaaacagct ggagctcttg gctcaagact
     3241 ataagctgcg aattaagcag attacggagg aagtggagag gcaggtgtcg actgcaatgg
     3301 ccgaggagat caggcgcctc tctgtactgg tggacgatta ccagatggac ttccaccctt
     3361 ctccagtagt cctcaaggtt tataagaatg agctgcaccg ccacatagag gaaggactgg
     3421 gtcgaaacat gtctgaccgc tgctccacgg ccatcaccaa ctccctgcag accatgcagc
     3481 aggacatgat agatggcttg aaacccctcc ttcctgtgtc cgtgcggagt cagatagaca
     3541 tgctggtccc acgccagtgc ttctccctca actatgacct aaactgtgac aagctgtgtg
     3601 ctgacttcca ggaagacatt gagttccatt tctctctcgg atggaccatg ctggtgaata
     3661 ggttcctggg ccccaagaac agccgtcggg ccttgatggg ctacaatgac caggtccagc
     3721 gtcccatccc tctgacgcca gccaacccca gcatgccccc actgccacag ggctcgctca
     3781 cccaggagga gttcatggtt tccatggtta ccggcctggc ctccttgaca tccaggacct
     3841 ccatgggcat tcttgttgtt ggaggagtgg tgtggaaggc agtgggctgg cggctcattg
     3901 ccctctcctt tgggctctat ggcctcctct acgtctatga gcgtctgacc tggaccacca
     3961 aggccaagga gagggccttc aagcgccagt ttgtggagca tgccagcgag aagctgcagc
     4021 ttgtcatcag ctacactggc tccaactgca gccaccaagt ccagcaggaa ctgtctggga
     4081 cctttgctca tctgtgtcag caagttgacg tcacccggga gaacctggag caggaaattg
     4141 ccgccatgaa caagaaaatt gaggttcttg actcacttca gagcaaagca aagctgctca
     4201 ggaataaagc cggttggttg gacagtgagc tcaacatgtt cacacaccag tacctgcagc
     4261 ccagcagata agcggccgct acgtaaattc cgcccccccc cccctctccc tccccccccc
     4321 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     4381 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     4441 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     4501 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     4561 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     4621 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     4681 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     4741 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     4801 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     4861 aacacgatga taatatggcc acaaccagat ctatgtccgt cctgacgccg ctgctgctgc
     4921 ggggcttgac aggctcggcc cggcggctcc cagtgccgcg cgccaagatc cattcgttgg
     4981 gggatccacc ggtcgccacc atggtgtcta agggcgaaga gctgattaag gagaacatgc
     5041 acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc acatccgagg
     5101 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc
     5161 ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagc agaaccttca
     5221 tcaaccacac ccagggcatc cccgatttct ttaagcagtc cttccctgag ggcttcacat
     5281 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
     5341 tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc ccatccaacg
     5401 gccctgtgat gcagaagaaa acactcggct gggaggccaa caccgagacc ctgtaccccg
     5461 ctgacggcgg cctggaaggc agaaccgaca tggccctgaa gctcgtgggc gggggccacc
     5521 tgatctgcaa cttcaagacc acatacagat ccaagaaacc cgctaagaac ctcaagatgc
     5581 ccggcgtcta ctatgtggac cacagactgg aaagaatcaa ggaggccgac aaagagacct
     5641 acgtcgagca gcacgaggtg gctgtggcca gatactgcga cctccctagc aaactggggc
     5701 acaaacttaa tggcatggac gagctgtaca agtaaagcgg ccgccagcac agtggtcgat
     5761 cgaggggggg cccgcgatta gtccaatttg ttaaagacag gatatcagtg gtccaggctc
     5821 tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca tagatagaat
     5881 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     5941 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     6001 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     6061 gtggtaagca gttcctgccc cggctcaggg ccaagaacag ttggaacagc agaatatggg
     6121 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     6181 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     6241 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     6301 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     6361 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt
     6421 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     6481 ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt
     6541 cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac gcgcggggag
     6601 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6661 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6721 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6781 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6841 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6901 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6961 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7021 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7081 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7141 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7201 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7261 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7321 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7381 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7441 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7501 tttgcggccg gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7561 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7621 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7681 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7741 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7801 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7861 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7921 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7981 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8041 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8101 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8161 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8221 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8281 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8341 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8401 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8461 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8521 cgcacatttc
//
