LOCUS       sequence-329883-        7609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKatet2 fused to TFAM dC-tail mutant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7609)
  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 7609)
  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_dC_mKate2
FEATURES             Location/Qualifiers
     source          1..7609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             450..1148
                     /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    1161..1749
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1214..1234)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1632..1643)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1823..2056
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2128..2249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2165..2184)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2219..2238
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2289..2424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2457..2473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2457..2473)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2470..2492)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2481..2497
                     /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(2505..2511,2512..2529,2530..2535))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2550..2571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2688..2705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2859..3447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3618..4409,4410..4478))
                     /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     4241..4260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4479..4583)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4651..4669
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4707..4729)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4829..4848
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5042..5064
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5056..5073
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5057..5073
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5088..5314
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5315..5495
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5542..5667
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6160..6393
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6578..6622
                     /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    6889..7005
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7095..7298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7249..7269
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7295..7319
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ttagaagaat tgcccagcgt tggagggaac ttcctgattc aaagaaaaaa atatatcaag
       61 atgcttatag ggcggagtgg caggtatata aagaagagat aagcagattt aaagaacagc
      121 taactccaag tcagattatg tctttggaaa aagaaatcat ggacaaacat ttaaaaagga
      181 aagctatgac aaaaaaaaaa gagttaacac tgcttggaaa accaaaaaga cctcgttcag
      241 cttataacgt ttatgtagct gaaagattcc aagaagctaa gggtgattca ccgcaggaaa
      301 agctgaagac tgtaaaggaa aactggaaaa atctgtctga ctctgaaaag gaattatata
      361 ttcagcatgc taaagaggac gaaactcgtt atcataatga aatgaagtct tgggaagaac
      421 aaatgggagg aagtggtgct agccccggga tggtgagcga gctgattaag gagaacatgc
      481 acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc acatccgagg
      541 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggcggtc gagggcggcc
      601 ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagc aaaaccttca
      661 tcaaccacac ccagggcatc cccgacttct ttaagcagtc cttccccgag ggcttcacat
      721 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
      781 tccgggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc ccatccaacg
      841 gccctgtgat gcagaagaaa acactcggct gggaggcctc caccgagacc ctgtaccccg
      901 ctgacggcgg cctggaaggc agagccgaca tggccctgaa gctcgtgggc gggggccacc
      961 tgatctgcaa cttgaagacc acatacagat ccaagaaacc cgctaagaac ctcaagatgc
     1021 ccggcgtcta ctatgtggac agaagactgg aaagaatcaa ggaggccgac aaagagacct
     1081 acgtcgagca gcacgaggtg gctgtggcca gatactgcga cctccctagc aaactggggc
     1141 acagatgaga attcgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     1201 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     1261 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     1321 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     1381 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     1441 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     1501 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     1561 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     1621 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     1681 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     1741 cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     1801 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag ataagatctg
     1861 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     1921 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     1981 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     2041 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     2101 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     2161 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2221 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     2281 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2341 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     2401 agtgaggagg cttttttgga ggcctagact tttgcagaga ccaaattcgt aatcatgtca
     2461 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2521 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2581 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2641 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2701 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2761 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2821 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2881 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2941 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3001 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3061 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3121 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3181 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3241 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3301 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3361 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3421 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3481 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3541 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3601 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3661 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3721 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3781 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3841 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3901 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3961 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4021 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4081 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4141 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4201 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4261 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4321 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4381 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4441 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4501 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4561 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     4621 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     4681 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     4741 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     4801 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4861 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     4921 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     4981 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     5041 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgacgcgtgt agtcttatgc
     5101 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
     5161 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
     5221 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
     5281 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
     5341 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     5401 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     5461 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
     5521 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
     5581 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
     5641 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
     5701 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
     5761 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     5821 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     5881 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     5941 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     6001 caagcggcca ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     6061 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     6121 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     6181 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     6241 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     6301 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     6361 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     6421 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     6481 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     6541 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     6601 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     6661 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     6721 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     6781 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     6841 agacagatcc attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa
     6901 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     6961 acaaactaaa gaattacaaa aacaaattac aaaattcaaa attttatcga tactagtatt
     7021 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     7081 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     7141 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     7201 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     7261 gtaggcgtgt acggtgggag gtttatataa gcagagctcg tttagtgaac cgtcagatcg
     7321 cctggagacg ccatccacgc tgttttgacc tccatagaag attctagaat ggcgtttctc
     7381 cgaagcatgt ggggcgtgct gagtgccctg ggaaggtctg gagcagagct gtgcaccggc
     7441 tgtggaagtc gactgcgctc ccccttcagt tttgtgtatt taccgaggtg gttttcatct
     7501 gtcttggcaa gttgtccaaa gaaacctgta agttcttacc ttcgattttc taaagaacaa
     7561 ctacccatat ttaaagctca gaacccagat gcaaaaacta cagaactaa
//
