LOCUS       sequence-330362-        7681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKate2 fused to full length TFAM in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7681)
  AUTHORS   Feric M, Demarest TG, Tian J, Croteau DL, Bohr VA, Misteli T
  TITLE     Self-assembly of multi-component mitochondrial nucleoids via phase 
            separation.
  JOURNAL   EMBO J. 2021 Mar 15;40(6):e107165. doi: 10.15252/embj.2020107165. 
            Epub 2021 Feb 23.
  PUBMED    33619770
REFERENCE   2  (bases 1 to 7681)
  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: "EMBO J."; 
            date: "2021-03-15"; volume: "40(6):e107165. doi"; pages: " 
            10.15252/embj.2020107165. Epub 2021 Feb 23"
COMMENT     Sequence Label: pCDH_CMV_TFAM_mKate2
FEATURES             Location/Qualifiers
     source          1..7681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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"
     misc_feature    1575..1691
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1781..1984
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1935..1955
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1981..2005
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2817..3515
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLRDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     misc_feature    3528..4116
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3581..3601)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3999..4010)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4190..4423
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4495..4616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4532..4551)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4586..4605
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4656..4791
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4824..4840)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4824..4840)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4837..4859)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4848..4864
                     /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(4872..4878,4879..4896,4897..4902))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4917..4938
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5055..5072)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5226..5814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5985..6776,6777..6845))
                     /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     6608..6627
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6846..6950)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7018..7036
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7074..7096)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7196..7215
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7409..7431
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7423..7440
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7424..7440
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7455..7681
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     1561 cggtatcggt taacttttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     1621 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     1681 ttcaaaattt tatcgatact agtattatgc ccagtacatg accttatggg actttcctac
     1741 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     1801 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     1861 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     1921 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag
     1981 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
     2041 tagaagattc tagaatggcg tttctccgaa gcatgtgggg cgtgctgagt gccctgggaa
     2101 ggtctggagc agagctgtgc accggctgtg gaagtcgact gcgctccccc ttcagttttg
     2161 tgtatttacc gaggtggttt tcatctgtct tggcaagttg tccaaagaaa cctgtaagtt
     2221 cttaccttcg attttctaaa gaacaactac ccatatttaa agctcagaac ccagatgcaa
     2281 aaactacaga actaattaga agaattgccc agcgttggag ggaacttcct gattcaaaga
     2341 aaaaaatata tcaagatgct tatagggcgg agtggcaggt atataaagaa gagataagca
     2401 gatttaaaga acagctaact ccaagtcaga ttatgtcttt ggaaaaagaa atcatggaca
     2461 aacatttaaa aaggaaagct atgacaaaaa aaaaagagtt aacactgctt ggaaaaccaa
     2521 aaagacctcg ttcagcttat aacgtttatg tagctgaaag attccaagaa gctaagggtg
     2581 attcaccgca ggaaaagctg aagactgtaa aggaaaactg gaaaaatctg tctgactctg
     2641 aaaaggaatt atatattcag catgctaaag aggacgaaac tcgttatcat aatgaaatga
     2701 agtcttggga agaacaaatg attgaagttg gacgaaagga tcttctacgt cgcacaataa
     2761 agaaacaacg aaaatatggt gctgaggagt gtggaggaag tggtgctagc cccgggatgg
     2821 tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca
     2881 accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     2941 tgagaatcaa ggcggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca
     3001 gcttcatgta cggcagcaaa accttcatca accacaccca gggcatcccc gacttcttta
     3061 agcagtcctt ccccgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     3121 tgctgaccgc tacccaggac accagcctcc gggacggctg cctcatctac aacgtcaaga
     3181 tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     3241 aggcctccac cgagaccctg taccccgctg acggcggcct ggaaggcaga gccgacatgg
     3301 ccctgaagct cgtgggcggg ggccacctga tctgcaactt gaagaccaca tacagatcca
     3361 agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggacaga agactggaaa
     3421 gaatcaagga ggccgacaaa gagacctacg tcgagcagca cgaggtggct gtggccagat
     3481 actgcgacct ccctagcaaa ctggggcaca gatgagaatt cgtcgacaat caacctctgg
     3541 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3601 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3661 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3721 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3781 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3841 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3901 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     3961 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     4021 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     4081 agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact
     4141 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     4201 attcactccc aacgaagata agatctgctt tttgcttgta ctgggtctct ctggttagac
     4261 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     4321 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     4381 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     4441 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     4501 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4561 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4621 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4681 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4741 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt
     4801 gcagagacca aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4861 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4921 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4981 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5041 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5101 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5161 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5221 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5281 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5341 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5401 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5461 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5521 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5581 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5641 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5701 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5761 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5821 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5881 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5941 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6001 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6061 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6121 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6181 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6241 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6301 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6361 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6421 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6481 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6541 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6601 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6661 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6721 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6781 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6841 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6901 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6961 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     7021 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     7081 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     7141 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     7201 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     7261 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     7321 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     7381 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     7441 gccaagctga cgcgtgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
     7501 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
     7561 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
     7621 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
     7681 c
//
