LOCUS       sequence-237768-        6258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mitoRFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6258)
  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 6258)
  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: pMYs-IRES-mitoRFP
FEATURES             Location/Qualifiers
     source          1..6258
                     /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"
     misc_feature    join(2028..2602,2603..2605,2606..2614)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2195..2212)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2422..2441
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2621..2695
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      2723..2732
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2729..3460
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTETLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     primer_bind     complement(4362..4379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4533..5121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4613..4632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5285..6076,6077..6145))
                     /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     5908..5927
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6146..6250)
                     /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 cgccaccgcg gccgctacgt aaattccgcc cccccccccc ctctccctcc
     2041 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     2101 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     2161 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     2221 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     2281 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     2341 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     2401 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     2461 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     2521 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     2581 ctttgaaaaa cacgatgata atatggccac aaccagatct atgtccgtcc tgacgccgct
     2641 gctgctgcgg ggcttgacag gctcggcccg gcggctccca gtgccgcgcg ccaagatcca
     2701 ttcgttgggg gatccaccgg tcgccaccat ggtgtctaag ggcgaagagc tgattaagga
     2761 gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac
     2821 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga
     2881 gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcag
     2941 aaccttcatc aaccacaccc agggcatccc cgatttcttt aagcagtcct tccctgaggg
     3001 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga
     3061 caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttccc
     3121 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccaaca ccgagaccct
     3181 gtaccccgct gacggcggcc tggaaggcag aaccgacatg gccctgaagc tcgtgggcgg
     3241 gggccacctg atctgcaact tcaagaccac atacagatcc aagaaacccg ctaagaacct
     3301 caagatgccc ggcgtctact atgtggacca cagactggaa agaatcaagg aggccgacaa
     3361 agagacctac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc tccctagcaa
     3421 actggggcac aaacttaatg gcatggacga gctgtacaag taaagcggcc gccagcacag
     3481 tggtcgatcg agggggggcc cgcgattagt ccaatttgtt aaagacagga tatcagtggt
     3541 ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata
     3601 gatagaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     3661 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg
     3721 agaatagaga agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca
     3781 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagtt ggaacagcag
     3841 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3901 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     3961 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     4021 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     4081 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa
     4141 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     4201 gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg
     4261 ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc
     4321 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4381 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4441 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4501 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4561 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4621 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4681 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4741 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4801 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4861 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4921 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4981 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5041 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5101 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5161 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5221 tagatccttt tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg
     5281 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5341 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5401 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5461 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5521 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5581 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5641 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5701 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5761 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5821 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5881 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5941 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6001 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6061 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6121 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6181 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6241 gggttccgcg cacatttc
//
