LOCUS       sequence-409480-        7694 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 7694)
  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 7694)
  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 deltaN380
FEATURES             Location/Qualifiers
     source          1..7694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     935..954
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(974..1051)
                     /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(1129..1151)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1324..1343
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      1381..1809
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1468..1487)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1678..1699
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1886..1903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2057..2645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2137..2156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2816..3607,3608..3676))
                     /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     3439..3458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3677..3768)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(3787..3814)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3906..3992)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4145..4162
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(7208..7228)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(7250..7267)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7289..7311)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7300..7316
                     /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(join(7585..7694,1..973))
                     /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 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
       61 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
      121 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
      181 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
      241 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
      301 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
      361 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
      421 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
      481 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
      541 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
      601 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
      661 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
      721 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
      781 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
      841 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
      901 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
      961 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     1021 cgcgaaaggt tttgcaccat tcgatggtgt caacgtaaat gcatgccgct tcgccttcgc
     1081 gcgcgaattg atctgctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1141 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1201 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     1261 cgatagcgga gtgtaccagt cacgtagcga tatcgctacg tgactggtac tggcttaact
     1321 atgcggcatc agagcagatt gtactgagag tgcaccatag cagcctgaat ggcgaatggg
     1381 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     1441 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     1501 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     1561 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     1621 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     1681 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     1741 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     1801 acgcgaattt taacaaaatt atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     1861 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1921 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1981 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2041 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2101 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2161 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2221 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2281 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2341 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2401 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2461 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     2521 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2581 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2641 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2701 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2761 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2821 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2881 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2941 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3001 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3061 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3121 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3181 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3241 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3301 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3361 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3421 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3481 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3541 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3601 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3661 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3721 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaaag agtttgtaga
     3781 aacgcaaaaa ggccatccgt caggatggcc ttctgcttaa tttgatgcct ggcagtttat
     3841 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttcg caacgttcaa atccgctccc
     3901 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     3961 cagtctttcg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcatg
     4021 gggagacccc acactaccat cggcgctacg gcgtttcact tctgagttcg gcatggggtc
     4081 aggtgggacc accgcgctac tgccgccagg caaattctgt tttatcagac cgcttctgcg
     4141 ttctgattta atctgtatca ggctgaaaat cttctctcat ccgccaaaac agccaagctt
     4201 ggtaccgcat gcctcgagac tgcaggctct agattcgaaa gcggccgcga ctagtgagct
     4261 ctcacgagaa aaaatattga ctgtttctca atactgtaac atcaaaatca cctttaggtg
     4321 ggatttcagg taatctcaac ggtaaaagaa cactcatacc gctattttca tctccaccgt
     4381 tttctagttc aagtgcgtcc aagtcagtga tgtcttcaaa taatgataca gttgtaacca
     4441 tcttttcacc aacatttcta tcttctatta gagggctgtt taataattct tgcctttttt
     4501 tctcaaaata aatttcatct gctaatgtag tacttcttcc taattttctg gataaatctt
     4561 tacggattct aataatcttt tgagccaaat ccgtagatct cttcaacttt ccaatcttga
     4621 cgtaattctt atacctttgg tcaaactcct ccttcaatct taaaaccaga tctacctcat
     4681 ggagtttgat aaactgttcc cttaacacca cattcatagt gtctatatca gatgcatgcg
     4741 tccagtaaga atcatgaact gatgcgaaat ctaatccttg ctttccacat tctgctgcac
     4801 ttagcaacat atgtgaagca tccagcgaat ggatgaagtt cggtggaaga cctgcctttt
     4861 gccttcttgc attaacaggg ttgacagcga aaggatcgct aataaagaca gtttgcaaat
     4921 tggtctcaac ttgctttttg ctttcttcac gatatggctg aacaattggt agtcctagtg
     4981 gtgtcgtcca tatgacggat gacataaagt cgggtttatt accgttcttg aacgattttt
     5041 catcaacatc caacctgata gatttggaaa ttcttttcgc actttcacct aaccaatcct
     5101 gaattaaatg cgcactatga aataactcgc ggatagcact aaaaacatgt ttggttaggt
     5161 atttagaaaa gtctaagctt tctttacgat catcaaatat tgggcttaat tgtttagcaa
     5221 tttgaaaagt agccccgacg taggtaacac cataaacgtt tgtcatgacg gtttgcttga
     5281 caacctttct ggtaatctta tccttcaaaa ttttggcgtt ttcgtctcct ttttctgctg
     5341 caatttctaa tcttttttgc accaaccttg caacatgagc ataaacatct tgtggtttat
     5401 cgcttggaac cagattcact tgagtagccc cttcaacgtc accacccaag gcagcataat
     5461 gttgcaaacc attgcaagta ccgtcttggt gaaccggttg atgagagata aactcctctg
     5521 gattatccat tttcataact tcattcaatt caaagcatgt ggccaatgct tgccatggct
     5581 tatcagcagt agtccaccaa cgatctcccg ttaagggatt ttccgcagaa tccttgatgt
     5641 cctgcaaatg agactcagta aatgcaacac gatcctttag gggtaactta tcaaatccga
     5701 acagatttga caaatgaatc tttaaccact tgagaccaga aggtcctaat tttttaccgt
     5761 gccaaaaaat caaaaggcca cgactcatat cattaccaag atgattgaaa tgtggagaca
     5821 aaggataagc acgtccccta aagtccaaat tatgtggaaa gtacagcttt tcacccaaaa
     5881 atgctcttgc aatttccaat ttgtagttcg tatcacatct atttgatcta tctgaagaga
     5941 atttattagc aattgttttg acttgaagtt tccaggctct caaaatggaa gggtcggagt
     6001 tcttcggagg agctggtggt aaaaccattt cgtcttgtgc acctggaata tctaggaaac
     6061 cttcaccttt attccaaact tgtgaaacca catcaaaaac ttttctgttg acagtccacg
     6121 gtgttctacc cagaacattc aaaccatcat aaacgcggtc tatatcacca ttatcggaag
     6181 cagctttcaa atatgctact tgttcgggtg agtcttttgt tcttaaaagg gtagattgtg
     6241 tataatgata tccacctgat ctccaattaa cccagggttt gggttccacc aacattggta
     6301 ataattgtgg ttggacagag gcaatcaatc tttcaccgtt caattgacga atcaaagtct
     6361 tgtgaatctt caagacaccc aatttggaac cattgtggta ctgataacca tgcgcaaaag
     6421 ctggagcttc accgtgaacc ttagcttttg tcacagggtc aacaccttgc acagatacct
     6481 tagcgacttg aatcaacatt gagattaata ccgaaccgat tctggcccta atactctgag
     6541 gccacaaaat tttcgactgc tcaatttgag atgatctgaa tacagattta gcattttgta
     6601 ccagtttttt gaattctggt gatttcttgt taacatctct gaacgcttga gattcgcttt
     6661 tcaaaacctg ctcggatcta aattccattt caatagcttt acccactgat ataaccgctc
     6721 ttgcggtcct cataccttca ataacaccac ccgtagaatt taactttaat agttccaaaa
     6781 tggtgatgac acacatttta ccagggtcaa tcaaggttaa gtatggtcca tatccaagac
     6841 ggttcgtgtc aactttattc aagcctttct tatctgatag cttttctgac aataatgacc
     6901 tacaatgatt aatttcttcc ttcactagag gtaacatttc attataccat ttccacaact
     6961 taacgtttaa attcttttca attgaaatgt cacctctagc ttttgcctct tcaaaatcgt
     7021 gcttccaacg ctctcttgcg gcttccgttg cacggttttc aagtaccttt tgtctatctt
     7081 ggttaaatat atttaatgcg ctttcgaatg ccttcttttc ctctaatgtt ggcagtgaat
     7141 tgtaaatttc aaaaaaattt atagaattat tattatcgtc aggcctttga attccggatc
     7201 ccatggcgcc ctgaaaatac aggttttcgg tcgttgggat atcgtaatcg tgatggtgat
     7261 ggtgatggta gtacgacatg gtctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7321 acagattata cggaccggat gattaattgt caacagctca tttcagaata tttgccagaa
     7381 ccgttatgat gtcggcgcaa aaaacattat ccagaacggg agtgcgcctt gagcgacacg
     7441 aattatgcag tgatttacga cctgcacagc cataccacag cttccgatgg ctgcctgacg
     7501 ccagaagcat tggtgcaccg tgcagtcgat gataagctgt caaaccagat caattcgcgc
     7561 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     7621 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggtgccag
     7681 ggtggttttt cttt
//
