LOCUS       sequence-329823-        5709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TFAM dHMGA mutant in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5709)
  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 5709)
  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_dHMGA_pET28
FEATURES             Location/Qualifiers
     source          1..5709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     963..982
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1002..1079)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1285..1304
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1388..1407
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1388..1406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1407..1431
                     /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             1446..1468
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1487..1504
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1514..1531
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1535..1567
                     /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"
     CDS             1961..1978
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2031..2049)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2045..2092
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2129..2584
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2216..2235)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2426..2447
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2676..3491)
                     /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(2676..3491)
                     /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     3399..3418
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3483..3502
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3613..4201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4102..4121
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4355..4372
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4387..4526
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4452..4471)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4612..4634
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4628..4819)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5628..5709,1..1001))
                     /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"
ORIGIN
        1 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
       61 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
      121 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
      181 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
      241 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
      301 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
      361 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
      421 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
      481 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
      541 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
      601 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
      661 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
      721 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
      781 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
      841 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
      901 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
      961 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     1021 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     1081 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     1141 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     1201 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     1261 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     1321 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     1381 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
     1441 ataattttgt ttaactttaa gaaggagata taccatgggc agcagccatc atcatcatca
     1501 tcacagcagc ggcctggtgc cgcgcggcag ccatatggct agcatgactg gtggacagca
     1561 aatgggtcgc ggatccatgc caagtcagat tatgtctttg gaaaaagaaa tcatggacaa
     1621 acatttaaaa aggaaagcta tgacaaaaaa aaaagagtta acactgcttg gaaaaccaaa
     1681 aagacctcgt tcagcttata acgtttatgt agctgaaaga ttccaagaag ctaagggtga
     1741 ttcaccgcag gaaaagctga agactgtaaa ggaaaactgg aaaaatctgt ctgactctga
     1801 aaaggaatta tatattcagc atgctaaaga ggacgaaact cgttatcata atgaaatgaa
     1861 gtcttgggaa gaacaaatga ttgaagttgg acgaaaggat cttctacgtc gcacaataaa
     1921 gaaacaacga aaatatggtg ctgaggagtg ttaactcgag caccaccacc accaccactg
     1981 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     2041 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     2101 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     2161 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2221 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2281 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2341 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2401 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2461 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2521 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     2581 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2641 tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa
     2701 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     2761 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     2821 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     2881 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     2941 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     3001 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     3061 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     3121 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     3181 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     3241 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     3301 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     3361 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     3421 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     3481 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     3541 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3601 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3661 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3721 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3781 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3841 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3901 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3961 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4021 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4081 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4141 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4201 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4261 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4321 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     4381 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     4441 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     4501 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     4561 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4621 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     4681 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     4741 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     4801 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     4861 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     4921 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     4981 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     5041 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     5101 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     5161 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     5221 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     5281 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     5341 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     5401 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     5461 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     5521 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     5581 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     5641 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5701 gcggtttgc
//
