LOCUS       sequence-409418-        7563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TFB2M protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7563)
  AUTHORS   Basu U, Lee SW, Deshpande A, Shen J, Sohn BK, Cho H, Kim H, Patel SS
  TITLE     The C-terminal tail of the yeast mitochondrial transcription factor 
            Mtf1 coordinates template strand alignment, DNA scrunching and 
            timely transition into elongation.
  JOURNAL   Nucleic Acids Res. 2020 Mar 18;48(5):2604-2620. doi: 
            10.1093/nar/gkaa040.
  PUBMED    31980825
REFERENCE   2  (bases 1 to 7563)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2020-03-18- 18"; volume: "48"; issue: 
            "5"; pages: "2604-2620"
COMMENT     Sequence Label: mt deltaN20 TFB2MdeltaC17
FEATURES             Location/Qualifiers
     source          1..7563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     89..108
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(128..205)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     411..430
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     514..533
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        514..532
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    533..557
                     /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             572..594
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             613..630
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             640..657
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             661..1761
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1735..1758
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1813..1833
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2941..2958
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3011..3029)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3025..3072
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3109..3564
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3196..3215)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3406..3427
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3656..4471)
                     /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(3656..4471)
                     /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     4379..4398
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4463..4482
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4593..5181
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5082..5101
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5335..5352
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5367..5506
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5432..5451)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5592..5614
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5608..5799)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6608..7563,1..127))
                     /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 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
       61 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
      121 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
      181 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
      241 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
      301 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
      361 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
      421 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
      481 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
      541 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
      601 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccac
      661 atgaaaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt
      721 ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat
      781 ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt
      841 atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc
      901 accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac
      961 aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa
     1021 gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg
     1081 aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg
     1141 ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa
     1201 gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt
     1261 aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa
     1321 ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa
     1381 gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt
     1441 ggtgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc
     1501 ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg
     1561 ggtgccgtag cgctgaagtc ttacgaggaa gagttggtga aagatccgcg gattgccgcc
     1621 actatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc
     1681 tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa
     1741 gccctgaaag acgcgcagac taattcgagc tcgaacaaca acaacaataa caataacaac
     1801 aacctcgggg atgaaaactt atatttccaa ggtgctagcg gtcgcttttg cattttaggg
     1861 tctgaagcgg cgacgcgaaa gcatttgccg gcgaggaacc actgtgggct ctctgactcc
     1921 tctccgcagc tgtggcccga accggatttc aggaatccgc caaggaaggc gtctaaggcc
     1981 agcttagact ttaagcgtta cgtaaccgat cggagattgg ctgagaccct ggcgcaaatc
     2041 tatttgggaa aaccaagtag acctccacac ctactgctgg agtgcaatcc aggtcctgga
     2101 atcctgactc aggcattact tgaagctggt gccaaagtgg ttgcgctcga aagtgacaaa
     2161 acttttattc cacatttgga gtccttagga aaaaatctgg atggaaaact acgagtgatt
     2221 cactgtgact tctttaaact agatcctaga agtggtggag taataaaacc acctgctatg
     2281 tcttctcgag ggctctttaa gaatttggga atagaagcag ttccttggac agcagacatc
     2341 cctttaaaag tagttggaat gttcccaagt agaggtgaga aaagggcact ttggaaactc
     2401 gcatatgact tgtattcctg tacttctata tataaatttg gacgaataga agtaaatatg
     2461 tttattggtg aaaaagaatt ccagaaacta atggcagatc ctggaaatcc agacttgtat
     2521 catgtattaa gtgttatctg gcaattagct tgtgagatta aggttctgca catggagcct
     2581 tggtcatcat ttgatatata cacccggaaa gggccgctgg aaaacccaaa gcgtagggaa
     2641 ttattagacc aattacaaca aaagctgtat cttattcaaa tgattcctcg tcaaaattta
     2701 tttaccaaga acttaacacc tatgaactat aatatatttt ttcacttgtt aaagcactgt
     2761 tttgggaggc gcagcgccac tgtaatagac cacttacgtt cattgactcc acttgatgcg
     2821 agagatatat tgatgcaaat aggaaaacag gaggatgaga aagtagttaa catgcaccct
     2881 caagacttca aaacactttt tgaaactata gagcgttaga agcttgcggc cgcactcgag
     2941 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     3001 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     3061 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     3121 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3181 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3241 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3301 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3361 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3421 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3481 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3541 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3601 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga
     3661 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     3721 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     3781 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     3841 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     3901 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     3961 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     4021 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     4081 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     4141 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     4201 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     4261 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     4321 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     4381 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag
     4441 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     4501 cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4561 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4621 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4681 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     4741 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4801 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4861 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4921 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4981 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5041 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5101 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5161 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5221 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5281 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5341 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     5401 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5461 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     5521 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5581 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5641 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     5701 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     5761 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     5821 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     5881 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     5941 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     6001 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     6061 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     6121 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     6181 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     6241 caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc
     6301 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     6361 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     6421 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     6481 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     6541 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     6601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     6721 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     6781 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     6841 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     6901 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     6961 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     7021 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     7081 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     7141 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     7201 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     7261 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     7321 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     7381 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     7441 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     7501 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     7561 ccg
//
