LOCUS       sequence-329666-        7309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKate2 fused to TFAM HMGA mutant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7309)
  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 7309)
  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_HMGA_mKate2
FEATURES             Location/Qualifiers
     source          1..7309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(4..108)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     176..194
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(232..254)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     354..373
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     567..589
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     581..598
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     582..598
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        613..839
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             840..1020
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1067..1192
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1685..1918
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2103..2147
                     /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    2414..2530
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2620..2823
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2774..2794
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2820..2844
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3284..3982
                     /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    3995..4583
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4048..4068)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4466..4477)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4657..4890
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4962..5083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4999..5018)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5053..5072
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5123..5258
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5291..5307)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5291..5307)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5304..5326)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5315..5331
                     /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(5339..5345,5346..5363,5364..5369))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5384..5405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5522..5539)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5693..6281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6452..7243,7244..7309,1..3))
                     /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     7075..7094
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
       61 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
      121 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
      181 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
      241 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
      301 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
      361 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
      421 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
      481 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
      541 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
      601 caagctgacg cgtgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga
      661 gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa
      721 ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa
      781 ccactgaatt gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg
      841 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
      901 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
      961 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     1021 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     1081 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     1141 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     1201 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     1261 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     1321 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     1381 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     1441 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     1501 aaacaaaagt aagaccaccg cacagcaagc ggccactgat cttcagacct ggaggaggag
     1561 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1621 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1681 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1741 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1801 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1861 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1921 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1981 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     2041 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     2101 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     2161 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     2221 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     2281 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2341 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
     2401 ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     2461 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat
     2521 tcaaaatttt atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact
     2581 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     2641 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     2701 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     2761 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga
     2821 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat
     2881 agaagattct agaatggcgt ttctccgaag catgtggggc gtgctgagtg ccctgggaag
     2941 gtctggagca gagctgtgca ccggctgtgg aagtcgactg cgctccccct tcagttttgt
     3001 gtatttaccg aggtggtttt catctgtctt ggcaagttgt ccaaagaaac ctgtaagttc
     3061 ttaccttcga ttttctaaag aacaactacc catatttaaa gctcagaacc cagatgcaaa
     3121 aactacagaa ctaattagaa gaattgccca gcgttggagg gaacttcctg attcaaagaa
     3181 aaaaatatat caagatgctt atagggcgga gtggcaggta tataaagaag agataagcag
     3241 atttaaagaa cagctaactg gaggaagtgg tgctagcccc gggatggtga gcgagctgat
     3301 taaggagaac atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa
     3361 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggc
     3421 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg
     3481 cagcaaaacc ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc
     3541 cgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     3601 ccaggacacc agcctccggg acggctgcct catctacaac gtcaagatca gaggggtgaa
     3661 cttcccatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg cctccaccga
     3721 gaccctgtac cccgctgacg gcggcctgga aggcagagcc gacatggccc tgaagctcgt
     3781 gggcgggggc cacctgatct gcaacttgaa gaccacatac agatccaaga aacccgctaa
     3841 gaacctcaag atgcccggcg tctactatgt ggacagaaga ctggaaagaa tcaaggaggc
     3901 cgacaaagag acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc
     3961 tagcaaactg gggcacagat gagaattcgt cgacaatcaa cctctggatt acaaaatttg
     4021 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4081 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4141 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4201 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4261 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4321 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4381 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     4441 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4501 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4561 ctccctttgg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     4621 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     4681 gaagataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     4741 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     4801 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     4861 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     4921 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     4981 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5041 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     5101 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5161 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5221 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat
     5281 tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5341 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5401 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5461 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5521 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5581 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5641 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5701 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5761 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5821 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5881 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5941 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6001 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6061 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6121 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6181 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6241 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6301 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6361 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6421 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6481 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6541 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6601 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6661 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6721 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6781 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6841 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6901 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6961 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7021 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7081 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7141 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7201 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7261 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatact
//
