LOCUS       sequence-330363-        7681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKate2 fused to TFAM L6 mutant 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_L6_mKate2
FEATURES             Location/Qualifiers
     source          1..7681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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    1802..1918
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2008..2211
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2162..2182
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2208..2232
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3044..3742
                     /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    3755..4343
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3808..3828)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4226..4237)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4417..4650
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4722..4843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4759..4778)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4813..4832
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4883..5018
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5051..5067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5051..5067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5064..5086)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5075..5091
                     /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(5099..5105,5106..5123,5124..5129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5144..5165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5282..5299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5453..6041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6212..7003,7004..7072))
                     /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     6835..6854
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7073..7177)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7245..7263
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7301..7323)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7423..7442
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7636..7658
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7650..7667
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7651..7667
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccactgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa
     1801 cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     1861 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaattc aaaattttat
     1921 cgatactagt attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     1981 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2041 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2101 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     2161 acgcaaatgg gcggtaggcg tgtacggtgg gaggtttata taagcagagc tcgtttagtg
     2221 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagattctag
     2281 aatggcgttt ctccgaagca tgtggggcgt gctgagtgcc ctgggaaggt ctggagcaga
     2341 gctgtgcacc ggctgtggaa gtcgactgcg ctcccccttc agttttgtgt atttaccgag
     2401 gtggttttca tctgtcttgg caagttgtcc aaagaaacct gtaagttctt accttcgatt
     2461 ttctaaagaa caactaccca tatttaaagc tcagaaccca gatgcaaaaa ctacagaact
     2521 aattagaaga attgcccagc gttggaggga acttcctgat tcaaagaaaa aaatatatca
     2581 agatgcttat agggcggagt ggcaggtata taaagaagag ataagcagat ttaaagaaca
     2641 gctaactcca agtcagatta tgtctttgga aaaagaaatc atggacgcag ctttagcagc
     2701 gaaagctatg acaaaagcag cagagttaac actgcttgga aaaccaaaaa gacctcgttc
     2761 agcttataac gtttatgtag ctgaaagatt ccaagaagct aagggtgatt caccgcagga
     2821 aaagctgaag actgtaaagg aaaactggaa aaatctgtct gactctgaaa aggaattata
     2881 tattcagcat gctaaagagg acgaaactcg ttatcataat gaaatgaagt cttgggaaga
     2941 acaaatgatt gaagttggac gaaaggatct tctacgtcgc acaataaaga aacaacgaaa
     3001 atatggtgct gaggagtgtg gaggaagtgg tgctagcccc gggatggtga gcgagctgat
     3061 taaggagaac atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa
     3121 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggc
     3181 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg
     3241 cagcaaaacc ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc
     3301 cgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     3361 ccaggacacc agcctccggg acggctgcct catctacaac gtcaagatca gaggggtgaa
     3421 cttcccatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg cctccaccga
     3481 gaccctgtac cccgctgacg gcggcctgga aggcagagcc gacatggccc tgaagctcgt
     3541 gggcgggggc cacctgatct gcaacttgaa gaccacatac agatccaaga aacccgctaa
     3601 gaacctcaag atgcccggcg tctactatgt ggacagaaga ctggaaagaa tcaaggaggc
     3661 cgacaaagag acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc
     3721 tagcaaactg gggcacagat gagaattcgt cgacaatcaa cctctggatt acaaaatttg
     3781 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3841 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3901 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3961 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4021 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4081 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4141 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     4201 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4261 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4321 ctccctttgg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     4381 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     4441 gaagataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     4501 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     4561 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     4621 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     4681 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     4741 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4801 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     4861 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4921 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4981 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat
     5041 tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5101 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5161 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5221 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5281 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5341 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5401 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5461 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5521 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5581 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5641 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5701 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5761 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5821 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5881 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5941 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6001 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6061 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6121 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6181 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6241 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6301 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6361 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6421 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6481 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6541 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6601 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6661 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6721 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6781 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6841 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6901 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6961 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7021 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7081 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7141 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7201 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     7261 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     7321 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     7381 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     7441 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     7501 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     7561 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     7621 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgacgc
     7681 g
//
