LOCUS       sequence-329882-        7354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKate2 fused to TFAM HMGB+C-tail mutant in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7354)
  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 7354)
  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_HMGB_ctail_mKate2_CLEAVAGE
FEATURES             Location/Qualifiers
     source          1..7354
                     /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             2490..3188
                     /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    3201..3789
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3254..3274)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3672..3683)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3863..4096
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4168..4289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4205..4224)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4259..4278
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4329..4464
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4497..4513)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4497..4513)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4510..4532)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4521..4537
                     /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(4545..4551,4552..4569,4570..4575))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4590..4611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4728..4745)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4899..5487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5658..6449,6450..6518))
                     /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     6281..6300
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6519..6623)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6691..6709
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6747..6769)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6869..6888
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7082..7104
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7096..7113
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7097..7113
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7128..7354
                     /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 tcaggaaaac caaaaagacc tcgttcagct tataacgttt
     2221 atgtagctga aagattccaa gaagctaagg gtgattcacc gcaggaaaag ctgaagactg
     2281 taaaggaaaa ctggaaaaat ctgtctgact ctgaaaagga attatatatt cagcatgcta
     2341 aagaggacga aactcgttat cataatgaaa tgaagtcttg ggaagaacaa atgattgaag
     2401 ttggacgaaa ggatcttcta cgtcgcacaa taaagaaaca acgaaaatat ggtgctgagg
     2461 agtgtggagg aagtggtgct agccccggga tggtgagcga gctgattaag gagaacatgc
     2521 acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc acatccgagg
     2581 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggcggtc gagggcggcc
     2641 ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagc aaaaccttca
     2701 tcaaccacac ccagggcatc cccgacttct ttaagcagtc cttccccgag ggcttcacat
     2761 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
     2821 tccgggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc ccatccaacg
     2881 gccctgtgat gcagaagaaa acactcggct gggaggcctc caccgagacc ctgtaccccg
     2941 ctgacggcgg cctggaaggc agagccgaca tggccctgaa gctcgtgggc gggggccacc
     3001 tgatctgcaa cttgaagacc acatacagat ccaagaaacc cgctaagaac ctcaagatgc
     3061 ccggcgtcta ctatgtggac agaagactgg aaagaatcaa ggaggccgac aaagagacct
     3121 acgtcgagca gcacgaggtg gctgtggcca gatactgcga cctccctagc aaactggggc
     3181 acagatgaga attcgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     3241 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3301 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3361 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3421 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3481 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3541 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     3601 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3661 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3721 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3781 cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     3841 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag ataagatctg
     3901 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     3961 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     4021 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     4081 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     4141 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     4201 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4261 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     4321 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4381 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4441 agtgaggagg cttttttgga ggcctagact tttgcagaga ccaaattcgt aatcatgtca
     4501 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4561 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4621 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4681 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4741 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4801 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4861 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4921 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4981 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5041 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5101 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5161 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5221 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5281 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5341 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5401 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5461 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5521 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5581 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5641 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5701 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5761 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5821 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5881 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5941 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6001 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6061 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6121 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6181 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6241 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6301 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6361 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6421 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6481 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6541 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6601 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6661 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     6721 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     6781 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     6841 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     6901 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     6961 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     7021 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     7081 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgacgcgtgt agtcttatgc
     7141 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
     7201 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
     7261 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
     7321 tattgtattt aagtgcctag ctcgatacat aaac
//
