LOCUS       sequence-329816-        5667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TFAM HMGA+linker mutant in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5667)
  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 5667)
  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: Human_TFAM_HMGA_linker_pET28
FEATURES             Location/Qualifiers
     source          1..5667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             209..226
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(279..297)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      293..340
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      377..832
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(464..483)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     674..695
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(924..1739)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(924..1739)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1647..1666
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1731..1750
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1861..2449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2350..2369
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2603..2620
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2635..2774
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2700..2719)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2860..2882
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2876..3067)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3876..4958)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4920..4939
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4959..5036)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5242..5261
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5345..5364
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5345..5363
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5364..5388
                     /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)."
     RBS             5403..5425
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5444..5461
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5471..5488
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5492..5524
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
ORIGIN
        1 gcgttggagg gaacttcctg attcaaagaa aaaaatatat caagatgctt atagggcgga
       61 gtggcaggta tataaagaag agataagcag atttaaagaa cagctaactc caagtcagat
      121 tatgtctttg gaaaaagaaa tcatggacaa acatttaaaa aggaaagcta tgacaaaaaa
      181 aaaagagtta acactgcttt aactcgagca ccaccaccac caccactgag atccggctgc
      241 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
      301 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
      361 cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
      421 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
      481 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
      541 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
      601 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
      661 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
      721 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
      781 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
      841 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      901 tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat
      961 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     1021 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     1081 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     1141 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
     1201 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     1261 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     1321 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     1381 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
     1441 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     1501 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     1561 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
     1621 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     1681 tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata
     1741 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc
     1801 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1861 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1921 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1981 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2041 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2101 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2161 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2221 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2281 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2341 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2401 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2461 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2521 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2581 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     2641 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     2701 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     2761 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2821 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     2881 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     2941 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3001 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     3061 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     3121 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     3181 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     3241 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     3301 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     3361 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     3421 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     3481 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg
     3541 ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga
     3601 aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg
     3661 cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta
     3721 cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc
     3781 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     3841 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3901 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3961 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     4021 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     4081 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     4141 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     4201 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     4261 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     4321 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     4381 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     4441 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     4501 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     4561 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     4621 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     4681 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     4741 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     4801 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     4861 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     4921 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     4981 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     5041 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     5101 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     5161 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     5221 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     5281 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
     5341 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
     5401 attttgttta actttaagaa ggagatatac catgggcagc agccatcatc atcatcatca
     5461 cagcagcggc ctggtgccgc gcggcagcca tatggctagc atgactggtg gacagcaaat
     5521 gggtcgcgga tccatgtcat ctgtcttggc aagttgtcca aagaaacctg taagttctta
     5581 ccttcgattt tctaaagaac aactacccat atttaaagct cagaacccag atgcaaaaac
     5641 tacagaacta attagaagaa ttgccca
//
