LOCUS       sequence-409473-        7946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RPO41 protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7946)
  AUTHORS   Paratkar S, Deshpande AP, Tang GQ, Patel SS
  TITLE     The N-terminal domain of the yeast mitochondrial RNA polymerase 
            regulates multiple steps of transcription.
  JOURNAL   J Biol Chem. 2011 May 6;286(18):16109-20. doi: 
            10.1074/jbc.M111.228023. Epub 2011 Mar 18.
  PUBMED    21454631
REFERENCE   2  (bases 1 to 7946)
  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: 
            "10.1074/jbc.M111.228023"; journalName: "J Biol Chem"; date: 
            "2011-05-6- 6"; volume: "286"; issue: "18"; pages: "16109-20"
COMMENT     Sequence Label: RPO41 deltaN296
FEATURES             Location/Qualifiers
     source          1..7946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(62..148)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     301..318
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(3616..3636)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(3658..3675)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3697..3719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3708..3724
                     /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)."
     CDS             complement(3993..5075)
                     /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     5037..5056
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5076..5153)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(5231..5253)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5426..5445
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      5483..5911
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5570..5589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5780..5801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5988..6005)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6159..6747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6239..6258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6918..7709,7710..7778))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7541..7560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7779..7870)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(7889..7916)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcgtcctgc ccgccaccct ccgggccgtt gcttcgcaac gttcaaatcc gctcccggcg
       61 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
      121 ctttcgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcatgggga
      181 gaccccacac taccatcggc gctacggcgt ttcacttctg agttcggcat ggggtcaggt
      241 gggaccaccg cgctactgcc gccaggcaaa ttctgtttta tcagaccgct tctgcgttct
      301 gatttaatct gtatcaggct gaaaatcttc tctcatccgc caaaacagcc aagcttggta
      361 ccgcatgcct cgagactgca ggctctagat tcgaaagcgg ccgcgactag tgagctctca
      421 cgagaaaaaa tattgactgt ttctcaatac tgtaacatca aaatcacctt taggtgggat
      481 ttcaggtaat ctcaacggta aaagaacact cataccgcta ttttcatctc caccgttttc
      541 tagttcaagt gcgtccaagt cagtgatgtc ttcaaataat gatacagttg taaccatctt
      601 ttcaccaaca tttctatctt ctattagagg gctgtttaat aattcttgcc tttttttctc
      661 aaaataaatt tcatctgcta atgtagtact tcttcctaat tttctggata aatctttacg
      721 gattctaata atcttttgag ccaaatccgt agatctcttc aactttccaa tcttgacgta
      781 attcttatac ctttggtcaa actcctcctt caatcttaaa accagatcta cctcatggag
      841 tttgataaac tgttccctta acaccacatt catagtgtct atatcagatg catgcgtcca
      901 gtaagaatca tgaactgatg cgaaatctaa tccttgcttt ccacattctg ctgcacttag
      961 caacatatgt gaagcatcca gcgaatggat gaagttcggt ggaagacctg ccttttgcct
     1021 tcttgcatta acagggttga cagcgaaagg atcgctaata aagacagttt gcaaattggt
     1081 ctcaacttgc tttttgcttt cttcacgata tggctgaaca attggtagtc ctagtggtgt
     1141 cgtccatatg acggatgaca taaagtcggg tttattaccg ttcttgaacg atttttcatc
     1201 aacatccaac ctgatagatt tggaaattct tttcgcactt tcacctaacc aatcctgaat
     1261 taaatgcgca ctatgaaata actcgcggat agcactaaaa acatgtttgg ttaggtattt
     1321 agaaaagtct aagctttctt tacgatcatc aaatattggg cttaattgtt tagcaatttg
     1381 aaaagtagcc ccgacgtagg taacaccata aacgtttgtc atgacggttt gcttgacaac
     1441 ctttctggta atcttatcct tcaaaatttt ggcgttttcg tctccttttt ctgctgcaat
     1501 ttctaatctt ttttgcacca accttgcaac atgagcataa acatcttgtg gtttatcgct
     1561 tggaaccaga ttcacttgag tagccccttc aacgtcacca cccaaggcag cataatgttg
     1621 caaaccattg caagtaccgt cttggtgaac cggttgatga gagataaact cctctggatt
     1681 atccattttc ataacttcat tcaattcaaa gcatgtggcc aatgcttgcc atggcttatc
     1741 agcagtagtc caccaacgat ctcccgttaa gggattttcc gcagaatcct tgatgtcctg
     1801 caaatgagac tcagtaaatg caacacgatc ctttaggggt aacttatcaa atccgaacag
     1861 atttgacaaa tgaatcttta accacttgag accagaaggt cctaattttt taccgtgcca
     1921 aaaaatcaaa aggccacgac tcatatcatt accaagatga ttgaaatgtg gagacaaagg
     1981 ataagcacgt cccctaaagt ccaaattatg tggaaagtac agcttttcac ccaaaaatgc
     2041 tcttgcaatt tccaatttgt agttcgtatc acatctattt gatctatctg aagagaattt
     2101 attagcaatt gttttgactt gaagtttcca ggctctcaaa atggaagggt cggagttctt
     2161 cggaggagct ggtggtaaaa ccatttcgtc ttgtgcacct ggaatatcta ggaaaccttc
     2221 acctttattc caaacttgtg aaaccacatc aaaaactttt ctgttgacag tccacggtgt
     2281 tctacccaga acattcaaac catcataaac gcggtctata tcaccattat cggaagcagc
     2341 tttcaaatat gctacttgtt cgggtgagtc ttttgttctt aaaagggtag attgtgtata
     2401 atgatatcca cctgatctcc aattaaccca gggtttgggt tccaccaaca ttggtaataa
     2461 ttgtggttgg acagaggcaa tcaatctttc accgttcaat tgacgaatca aagtcttgtg
     2521 aatcttcaag acacccaatt tggaaccatt gtggtactga taaccatgcg caaaagctgg
     2581 agcttcaccg tgaaccttag cttttgtcac agggtcaaca ccttgcacag ataccttagc
     2641 gacttgaatc aacattgaga ttaataccga accgattctg gccctaatac tctgaggcca
     2701 caaaattttc gactgctcaa tttgagatga tctgaataca gatttagcat tttgtaccag
     2761 ttttttgaat tctggtgatt tcttgttaac atctctgaac gcttgagatt cgcttttcaa
     2821 aacctgctcg gatctaaatt ccatttcaat agctttaccc actgatataa ccgctcttgc
     2881 ggtcctcata ccttcaataa caccacccgt agaatttaac tttaatagtt ccaaaatggt
     2941 gatgacacac attttaccag ggtcaatcaa ggttaagtat ggtccatatc caagacggtt
     3001 cgtgtcaact ttattcaagc ctttcttatc tgatagcttt tctgacaata atgacctaca
     3061 atgattaatt tcttccttca ctagaggtaa catttcatta taccatttcc acaacttaac
     3121 gtttaaattc ttttcaattg aaatgtcacc tctagctttt gcctcttcaa aatcgtgctt
     3181 ccaacgctct cttgcggctt ccgttgcacg gttttcaagt accttttgtc tatcttggtt
     3241 aaatatattt aatgcgcttt cgaatgcctt cttttcctct aatgttggca gtgaattgta
     3301 aatttcaaaa aaatttatag aattattatt atcgtcgttt ttcgtaggtt tcattttcaa
     3361 aacattatca ttggcatcaa atttgaattt agaaatttgc tctttctgtt ccggcgttaa
     3421 tgataatccc agcaaagtat gccggattac tctcattcca atagtgtcta cagctataag
     3481 cttttcagcg tcttttgaca agacttctgt tgaaccgtgg aatgaaactt cccgaagagg
     3541 atccaaggaa ggttttttta tagaatcaga gcttgcattt gatgctttag gcctttgaat
     3601 tccggatccc atggcgccct gaaaatacag gttttcggtc gttgggatat cgtaatcgtg
     3661 atggtgatgg tgatggtagt acgacatggt ctgtttcctg tgtgaaattg ttatccgctc
     3721 acaattccac agattatacg gaccggatga ttaattgtca acagctcatt tcagaatatt
     3781 tgccagaacc gttatgatgt cggcgcaaaa aacattatcc agaacgggag tgcgccttga
     3841 gcgacacgaa ttatgcagtg atttacgacc tgcacagcca taccacagct tccgatggct
     3901 gcctgacgcc agaagcattg gtgcaccgtg cagtcgatga taagctgtca aaccagatca
     3961 attcgcgcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4021 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4081 ggtgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     4141 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     4201 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     4261 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     4321 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     4381 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     4441 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     4501 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     4561 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     4621 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     4681 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     4741 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     4801 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     4861 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     4921 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     4981 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     5041 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     5101 tccgggcgct atcatgccat accgcgaaag gttttgcacc attcgatggt gtcaacgtaa
     5161 atgcatgccg cttcgccttc gcgcgcgaat tgatctgctg cctcgcgcgt ttcggtgatg
     5221 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     5281 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
     5341 cagccatgac ccagtcacgt agcgatagcg gagtgtacca gtcacgtagc gatatcgcta
     5401 cgtgactggt actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     5461 agcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5521 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5581 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5641 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     5701 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5761 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5821 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5881 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa ttatgcggtg tgaaataccg
     5941 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     6001 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6061 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6121 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6181 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6241 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6301 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6361 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6421 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6481 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6541 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6601 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6661 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6721 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6781 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6841 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6901 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6961 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7021 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7081 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7141 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7201 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7261 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7321 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7381 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7441 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7501 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     7561 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7621 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7681 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7741 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7801 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7861 aataaacaaa agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     7921 aatttgatgc ctggcagttt atggcg
//
