LOCUS       sequence-409419-        7605 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 7605)
  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 7605)
  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 TFB2MdeltaC3
FEATURES             Location/Qualifiers
     source          1..7605
                     /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             2983..3000
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3053..3071)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3067..3114
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3151..3606
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3238..3257)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3448..3469
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3698..4513)
                     /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(3698..4513)
                     /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     4421..4440
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4505..4524
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4635..5223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5124..5143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5377..5394
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5409..5548
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5474..5493)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5634..5656
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5650..5841)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6650..7605,1..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 gagcgttcca aagattgtgc ttataaatgg
     2941 ctgtatgatg aaaccctgta gaagcttgcg gccgcactag agcaccacca ccaccaccac
     3001 tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     3061 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     3121 ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     3181 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3241 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3301 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3361 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3421 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3481 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3541 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     3601 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3661 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
     3721 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     3781 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     3841 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     3901 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     3961 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     4021 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     4081 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     4141 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     4201 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     4261 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     4321 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     4381 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     4441 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     4501 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     4561 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4621 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaac
     4681 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4741 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     4801 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4861 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4921 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4981 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5041 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5101 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5161 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5221 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5281 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5341 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5401 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     5461 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     5521 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     5581 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5641 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     5701 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     5761 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     5821 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     5881 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     5941 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     6001 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     6061 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     6121 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     6181 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     6241 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     6301 cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
     6361 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
     6421 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
     6481 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
     6541 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     6601 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     6661 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6721 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     6781 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     6841 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     6901 cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg
     6961 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     7021 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     7081 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     7141 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     7201 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     7261 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     7321 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     7381 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     7441 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     7501 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     7561 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccg
//
