LOCUS       sequence-330358-        7444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKate2 fused to TFAM dHMGA mutant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7444)
  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 7444)
  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_dHMGA_mKate2_CLEAVAGE
FEATURES             Location/Qualifiers
     source          1..7444
                     /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             2580..3278
                     /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    3291..3879
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3344..3364)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3762..3773)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3953..4186
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4258..4379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4295..4314)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4349..4368
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4419..4554
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4587..4603)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4587..4603)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4600..4622)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4611..4627
                     /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(4635..4641,4642..4659,4660..4665))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4680..4701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4818..4835)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4989..5577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5748..6539,6540..6608))
                     /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     6371..6390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6609..6713)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6781..6799
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6837..6859)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6959..6978
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7172..7194
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7186..7203
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7187..7203
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7218..7444
                     /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 tcaccaagtc agattatgtc tttggaaaaa gaaatcatgg
     2221 acaaacattt aaaaaggaaa gctatgacaa aaaaaaaaga gttaacactg cttggaaaac
     2281 caaaaagacc tcgttcagct tataacgttt atgtagctga aagattccaa gaagctaagg
     2341 gtgattcacc gcaggaaaag ctgaagactg taaaggaaaa ctggaaaaat ctgtctgact
     2401 ctgaaaagga attatatatt cagcatgcta aagaggacga aactcgttat cataatgaaa
     2461 tgaagtcttg ggaagaacaa atgattgaag ttggacgaaa ggatcttcta cgtcgcacaa
     2521 taaagaaaca acgaaaatat ggtgctgagg agtgtggagg aagtggtgct agccccggga
     2581 tggtgagcga gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtga
     2641 acaaccacca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga
     2701 ccatgagaat caaggcggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta
     2761 ccagcttcat gtacggcagc aaaaccttca tcaaccacac ccagggcatc cccgacttct
     2821 ttaagcagtc cttccccgag ggcttcacat gggagagagt caccacatac gaagacgggg
     2881 gcgtgctgac cgctacccag gacaccagcc tccgggacgg ctgcctcatc tacaacgtca
     2941 agatcagagg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa acactcggct
     3001 gggaggcctc caccgagacc ctgtaccccg ctgacggcgg cctggaaggc agagccgaca
     3061 tggccctgaa gctcgtgggc gggggccacc tgatctgcaa cttgaagacc acatacagat
     3121 ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac agaagactgg
     3181 aaagaatcaa ggaggccgac aaagagacct acgtcgagca gcacgaggtg gctgtggcca
     3241 gatactgcga cctccctagc aaactggggc acagatgaga attcgtcgac aatcaacctc
     3301 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3361 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3421 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3481 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3541 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     3601 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3661 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     3721 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     3781 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     3841 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg
     3901 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     3961 ctaattcact cccaacgaag ataagatctg ctttttgctt gtactgggtc tctctggtta
     4021 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     4081 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     4141 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     4201 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac
     4261 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4321 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4381 catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4441 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4501 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact
     4561 tttgcagaga ccaaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg
     4621 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4681 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4741 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4801 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4861 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4921 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4981 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5041 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5101 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5161 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5221 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5281 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5341 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5401 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5461 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5521 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5581 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5641 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5701 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5761 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5821 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5881 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5941 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6001 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6061 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6121 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6181 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6241 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6301 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6361 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6421 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6481 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6541 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6601 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6661 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6721 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     6781 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     6841 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     6901 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     6961 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     7021 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     7081 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     7141 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     7201 agtgccaagc tgacgcgtgt agtcttatgc aatactcttg tagtcttgca acatggtaac
     7261 gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga
     7321 agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg
     7381 acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat
     7441 aaac
//
