LOCUS       sequence-329806-        7399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKate2 fused to TFAM HMGA+linker mutant in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7399)
  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 7399)
  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_linker_mKate2
FEATURES             Location/Qualifiers
     source          1..7399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(623..647)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(644..847)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(673..693)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    937..1053
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1320..1364)
                     /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    1549..1782
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2275..2400
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2447..2627
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2628..2854)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(2869..2886)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2869..2885)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2878..2900)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3094..3113)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3213..3235
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3273..3291)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3359..3463
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3464..3532,3533..4324)
                     /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     complement(3682..3701)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4495..5083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5237..5254
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5371..5392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5407..5412,5413..5430,5431..5437)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5445..5461
                     /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)."
     primer_bind     5450..5472
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5469..5485
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5469..5485
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      5518..5653
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5693..5814
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5704..5723)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5758..5777
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             5886..6119
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    6193..6781
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6299..6310
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6708..6728
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(6794..7399,1..93))
                     /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"
ORIGIN
        1 gccctcggat gtgcacttga agtggtggtt gttcacggtg ccctccatgt acagcttcat
       61 gtgcatgttc tccttaatca gctcgctcac catcccgggg ctagcaccac ttcctccaag
      121 cagtgttaac tctttttttt ttgtcatagc tttccttttt aaatgtttgt ccatgatttc
      181 tttttccaaa gacataatct gacttggagt tagctgttct ttaaatctgc ttatctcttc
      241 tttatatacc tgccactccg ccctataagc atcttgatat atttttttct ttgaatcagg
      301 aagttccctc caacgctggg caattcttct aattagttct gtagtttttg catctgggtt
      361 ctgagcttta aatatgggta gttgttcttt agaaaatcga aggtaagaac ttacaggttt
      421 ctttggacaa cttgccaaga cagatgaaaa ccacctcggt aaatacacaa aactgaaggg
      481 ggagcgcagt cgacttccac agccggtgca cagctctgct ccagaccttc ccagggcact
      541 cagcacgccc cacatgcttc ggagaaacgc cattctagaa tcttctatgg aggtcaaaac
      601 agcgtggatg gcgtctccag gcgatctgac ggttcactaa acgagctctg cttatataaa
      661 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
      721 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
      781 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
      841 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
      901 ataaggtcat gtactgggca taatactagt atcgataaaa ttttgaattt tgtaatttgt
      961 ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct ttcccctgca
     1021 ctgtaccccc caatcccccc ttttctttta aaagttaacc gataccgtcg agatccgttc
     1081 actaatcgaa tggatctgtc tctgtctctc tctccacctt cttcttctat tccttcgggc
     1141 ctgtcgggtc ccctcggggt tgggaggtgg gtctgaaacg ataatggtga atatccctgc
     1201 ctaactctat tcactataga aagtacagca aaaactattc ttaaacctac caagcctcct
     1261 actatcatta tgaataattt tatataccac agccaatttg ttatgttaaa ccaattccac
     1321 aaacttgccc atttatctaa ttccaataat tcttgttcat tcttttcttg ctggttttgc
     1381 gattcttcaa ttaaggagtg tattaagctt gtgtaattgt taatttctct gtcccactcc
     1441 atccaggtcg tgtgattcca aatctgttcc agagatttat tactccaact agcattccaa
     1501 ggcacagcag tggtgcaaat gagttttcca gagcaacccc aaatccccag gagctgttga
     1561 tcctttaggt atctttccac agccaggatt cttgcctgga gctgcttgat gccccagact
     1621 gtgagttgca acagatgctg ttgcgcctca atagccctca gcaaattgtt ctgctgctgc
     1681 actataccag acaataattg tctggcctgt accgtcagcg tcattgacgc tgcgcccata
     1741 gtgcttcctg ctgctcccaa gaacccaagg aacaaagctc ctattcccac tgctcttttt
     1801 tctctctgca ccactcttct ctttgccttg gtgggtgcta ctcctaatgg ttcaattttt
     1861 actactttat atttatataa ttcacttctc caattgtccc tcatatctcc tcctccaggt
     1921 ctgaagatca gtggccgctt gctgtgcggt ggtcttactt ttgttttgct cttcctctat
     1981 cttgtctaaa gcttccttgg tgtcttttat ctctatcctt tgatgcacac aatagagggt
     2041 tgctactgta ttatataatg atctaagttc ttctgatcct gtctgaaggg atggttgtag
     2101 ctgtcccagt atttgtctac agccttctga tgtttctaac aggccaggat taactgcgaa
     2161 tcgttctagc tccctgcttg cccatactat atgttttaat ttatattttt tctttccccc
     2221 tggccttaac cgaatttttt cccatcgcga tctaattctc ccccgcttaa tactgacgct
     2281 ctcgcaccca tctctctcct tctagcctcc gctagtcaaa atttttggcg tactcaccag
     2341 tcgccgcccc tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc gtcgagagag
     2401 ctcctctggt ttccctttcg ctttcaagtc cctgttcggg cgccactgct agagattttc
     2461 cacactgact aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca
     2521 cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt
     2581 tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccgtt tatgtatcga
     2641 gctaggcact taaatacaat atctctgcaa tgcggcaatt cagtggttcg tccaatccat
     2701 gtcagacccg tctgttgcct tcctaataag gcacgatcgt accaccttac ttccaccaat
     2761 cggcatgcac ggtgcttttt ctctccttgt aaggcatgtt gctaactcat cgttaccatg
     2821 ttgcaagact acaagagtat tgcataagac tacacgcgtc agcttggcac tggccgtcgt
     2881 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     2941 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     3001 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     3061 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3121 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3181 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3241 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3301 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3361 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3421 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3481 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3541 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3601 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3661 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3721 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3781 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3841 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3901 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3961 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4021 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4081 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4141 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4201 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4261 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4321 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4381 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4441 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4501 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4561 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4621 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4681 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4741 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4801 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4861 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4921 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5041 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5101 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5161 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5221 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     5281 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     5341 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     5401 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     5461 atttcacaca ggaaacagct atgacatgat tacgaatttg gtctctgcaa aagtctaggc
     5521 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     5581 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     5641 ttaggggcgg gatgggcgga gttaggggcg ggatagctag agccagacat gataagatac
     5701 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     5761 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttcctctc
     5821 actctctgat attcatttct ttgcaagtta taaatactga ataataagat gacatgaact
     5881 actactgcta gagattttcc acactgacta aaagggtctg agggatctct agttaccaga
     5941 gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt
     6001 aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag
     6061 agacccagta caagcaaaaa gcagatctta tcttcgttgg gagtgaatta gcccttccag
     6121 tccccccttt tcttttaaaa agtggctaag atctacagct gccttgtaag tcattggtct
     6181 taaaggtacc aggcggggag gcggcccaaa gggagatccg actcgtctga gggcgaaggc
     6241 gaagacgcgg aagaggccgc agagccggca gcaggccgcg ggaaggaagg tccgctggat
     6301 tgagggccga agggacgtag cagaaggacg tcccgcgcag aatccaggtg gcaacacagg
     6361 cgagcagcca aggaaaggac gatgatttcc ccgacaacac cacggaattg tcagtgccca
     6421 acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa
     6481 tagggagggg gaaagcgaaa gtcccggaaa ggagctgaca ggtggtggca atgccccaac
     6541 cagtgggggt tgcgtcagca aacacagtgc acaccacgcc acgttgcctg acaacgggcc
     6601 acaactcctc ataaagagac agcaaccagg atttatacaa ggaggagaaa atgaaagcca
     6661 tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag
     6721 gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat
     6781 tgtcgacgaa ttctcatctg tgccccagtt tgctagggag gtcgcagtat ctggccacag
     6841 ccacctcgtg ctgctcgacg taggtctctt tgtcggcctc cttgattctt tccagtcttc
     6901 tgtccacata gtagacgccg ggcatcttga ggttcttagc gggtttcttg gatctgtatg
     6961 tggtcttcaa gttgcagatc aggtggcccc cgcccacgag cttcagggcc atgtcggctc
     7021 tgccttccag gccgccgtca gcggggtaca gggtctcggt ggaggcctcc cagccgagtg
     7081 ttttcttctg catcacaggg ccgttggatg ggaagttcac ccctctgatc ttgacgttgt
     7141 agatgaggca gccgtcccgg aggctggtgt cctgggtagc ggtcagcacg cccccgtctt
     7201 cgtatgtggt gactctctcc catgtgaagc cctcggggaa ggactgctta aagaagtcgg
     7261 ggatgccctg ggtgtggttg atgaaggttt tgctgccgta catgaagctg gtagccagga
     7321 tgtcgaaggc gaaggggaga gggccgccct cgaccgcctt gattctcatg gtctgggtgc
     7381 cctcgtaggg cttgccttc
//
