LOCUS       sequence-421636-        8534 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 8534)
  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 8534)
  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 deltaN100
FEATURES             Location/Qualifiers
     source          1..8534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     448..467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     701..718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(795..1223)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(905..926)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1117..1136
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(1261..1280)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1453..1475
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        1553..1630
                     /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."
     CDS             1631..2713
                     /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     complement(1650..1669)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    2982..2998
                     /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)."
     primer_bind     2987..3009
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             3031..3048
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3070..3090
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(6976..6993)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      7146..7232
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7324..7351
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        7370..7461
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7462..7530,7531..8322)
                     /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     complement(7680..7699)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8493..8534,1..547)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
       61 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
      121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      601 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      661 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      721 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      781 cataattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
      841 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
      901 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
      961 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     1021 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     1081 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     1141 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     1201 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgctatggt gcactctcag
     1261 tacaatctgc tctgatgccg catagttaag ccagtaccag tcacgtagcg atatcgctac
     1321 gtgactggta cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     1381 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     1441 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     1501 cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca tgcatttacg ttgacaccat
     1561 cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     1621 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     1681 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     1741 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     1801 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     1861 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     1921 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     1981 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     2041 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     2101 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     2161 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     2221 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     2281 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     2341 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     2401 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     2461 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     2521 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     2581 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcacccaata cgcaaaccgc
     2641 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     2701 aagcgggcag tgagcgcaac gcaattaatg taagttagcg cgaattgatc tggtttgaca
     2761 gcttatcatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg
     2821 tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc
     2881 gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga
     2941 gctgttgaca attaatcatc cggtccgtat aatctgtgga attgtgagcg gataacaatt
     3001 tcacacagga aacagaccat gtcgtactac catcaccatc accatcacga ttacgatatc
     3061 ccaacgaccg aaaacctgta ttttcagggc gccatgggat ccggaattca aaggccttca
     3121 gcagtaacct cgatgacaag aactagagac gttatgcaat tatggtctct gttagaagca
     3181 tgcttgcaat cgaatctaat gaaaagagcg ttctctatat tggagtcact gtacctggta
     3241 cctgaacata agcagcgatt tatcgaggat tataacatgt acttgaattc attttccaaa
     3301 aatgacccta atttccctat tttaaaaatg aacgaaaaat taacaaatga tttggagaca
     3361 agttttaaag acgttaatta taatgacaaa actttggcaa taatgattca tcacgctttg
     3421 aacttccatt caacgacaag ctcgatgctt ttgaagccta taatctctgc ctacttgaaa
     3481 atgagcgtga atggtattag ggaaattttc tcctgcctag atattttgac catttcagat
     3541 ctgaatattt taatgaatga cttaaaggtt ataacaccat cgcaactgcc aaattctgta
     3601 agaccaatat tagaatcact tacgctatct cctacacctg taaataatat tgaaaatgag
     3661 gagggtttga acaaagtgga agctgagaac gattcaaaat tgcataaagc atcaaatgca
     3721 agctctgatt ctataaaaaa accttccttg gatcctcttc gggaagtttc attccacggt
     3781 tcaacagaag tcttgtcaaa agacgctgaa aagcttatag ctgtagacac tattggaatg
     3841 agagtaatcc ggcatacttt gctgggatta tcattaacgc cggaacagaa agagcaaatt
     3901 tctaaattca aatttgatgc caatgataat gttttgaaaa tgaaacctac gaaaaacgac
     3961 gataataata attctataaa tttttttgaa atttacaatt cactgccaac attagaggaa
     4021 aagaaggcat tcgaaagcgc attaaatata tttaaccaag atagacaaaa ggtacttgaa
     4081 aaccgtgcaa cggaagccgc aagagagcgt tggaagcacg attttgaaga ggcaaaagct
     4141 agaggtgaca tttcaattga aaagaattta aacgttaagt tgtggaaatg gtataatgaa
     4201 atgttacctc tagtgaagga agaaattaat cattgtaggt cattattgtc agaaaagcta
     4261 tcagataaga aaggcttgaa taaagttgac acgaaccgtc ttggatatgg accatactta
     4321 accttgattg accctggtaa aatgtgtgtc atcaccattt tggaactatt aaagttaaat
     4381 tctacgggtg gtgttattga aggtatgagg accgcaagag cggttatatc agtgggtaaa
     4441 gctattgaaa tggaatttag atccgagcag gttttgaaaa gcgaatctca agcgttcaga
     4501 gatgttaaca agaaatcacc agaattcaaa aaactggtac aaaatgctaa atctgtattc
     4561 agatcatctc aaattgagca gtcgaaaatt ttgtggcctc agagtattag ggccagaatc
     4621 ggttcggtat taatctcaat gttgattcaa gtcgctaagg tatctgtgca aggtgttgac
     4681 cctgtgacaa aagctaaggt tcacggtgaa gctccagctt ttgcgcatgg ttatcagtac
     4741 cacaatggtt ccaaattggg tgtcttgaag attcacaaga ctttgattcg tcaattgaac
     4801 ggtgaaagat tgattgcctc tgtccaacca caattattac caatgttggt ggaacccaaa
     4861 ccctgggtta attggagatc aggtggatat cattatacac aatctaccct tttaagaaca
     4921 aaagactcac ccgaacaagt agcatatttg aaagctgctt ccgataatgg tgatatagac
     4981 cgcgtttatg atggtttgaa tgttctgggt agaacaccgt ggactgtcaa cagaaaagtt
     5041 tttgatgtgg tttcacaagt ttggaataaa ggtgaaggtt tcctagatat tccaggtgca
     5101 caagacgaaa tggttttacc accagctcct ccgaagaact ccgacccttc cattttgaga
     5161 gcctggaaac ttcaagtcaa aacaattgct aataaattct cttcagatag atcaaataga
     5221 tgtgatacga actacaaatt ggaaattgca agagcatttt tgggtgaaaa gctgtacttt
     5281 ccacataatt tggactttag gggacgtgct tatcctttgt ctccacattt caatcatctt
     5341 ggtaatgata tgagtcgtgg ccttttgatt ttttggcacg gtaaaaaatt aggaccttct
     5401 ggtctcaagt ggttaaagat tcatttgtca aatctgttcg gatttgataa gttaccccta
     5461 aaggatcgtg ttgcatttac tgagtctcat ttgcaggaca tcaaggattc tgcggaaaat
     5521 cccttaacgg gagatcgttg gtggactact gctgataagc catggcaagc attggccaca
     5581 tgctttgaat tgaatgaagt tatgaaaatg gataatccag aggagtttat ctctcatcaa
     5641 ccggttcacc aagacggtac ttgcaatggt ttgcaacatt atgctgcctt gggtggtgac
     5701 gttgaagggg ctactcaagt gaatctggtt ccaagcgata aaccacaaga tgtttatgct
     5761 catgttgcaa ggttggtgca aaaaagatta gaaattgcag cagaaaaagg agacgaaaac
     5821 gccaaaattt tgaaggataa gattaccaga aaggttgtca agcaaaccgt catgacaaac
     5881 gtttatggtg ttacctacgt cggggctact tttcaaattg ctaaacaatt aagcccaata
     5941 tttgatgatc gtaaagaaag cttagacttt tctaaatacc taaccaaaca tgtttttagt
     6001 gctatccgcg agttatttca tagtgcgcat ttaattcagg attggttagg tgaaagtgcg
     6061 aaaagaattt ccaaatctat caggttggat gttgatgaaa aatcgttcaa gaacggtaat
     6121 aaacccgact ttatgtcatc cgtcatatgg acgacaccac taggactacc aattgttcag
     6181 ccatatcgtg aagaaagcaa aaagcaagtt gagaccaatt tgcaaactgt ctttattagc
     6241 gatcctttcg ctgtcaaccc tgttaatgca agaaggcaaa aggcaggtct tccaccgaac
     6301 ttcatccatt cgctggatgc ttcacatatg ttgctaagtg cagcagaatg tggaaagcaa
     6361 ggattagatt tcgcatcagt tcatgattct tactggacgc atgcatctga tatagacact
     6421 atgaatgtgg tgttaaggga acagtttatc aaactccatg aggtagatct ggttttaaga
     6481 ttgaaggagg agtttgacca aaggtataag aattacgtca agattggaaa gttgaagaga
     6541 tctacggatt tggctcaaaa gattattaga atccgtaaag atttatccag aaaattagga
     6601 agaagtacta cattagcaga tgaaatttat tttgagaaaa aaaggcaaga attattaaac
     6661 agccctctaa tagaagatag aaatgttggt gaaaagatgg ttacaactgt atcattattt
     6721 gaagacatca ctgacttgga cgcacttgaa ctagaaaacg gtggagatga aaatagcggt
     6781 atgagtgttc ttttaccgtt gagattacct gaaatcccac ctaaaggtga ttttgatgtt
     6841 acagtattga gaaacagtca atattttttc tcgtgagagc tcactagtcg cggccgcttt
     6901 cgaatctaga gcctgcagtc tcgaggcatg cggtaccaag cttggctgtt ttggcggatg
     6961 agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca
     7021 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt
     7081 gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca
     7141 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     7201 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
     7261 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta
     7321 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt
     7381 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     7441 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     7501 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     7561 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     7621 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     7681 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     7741 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     7801 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     7861 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7921 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     7981 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     8041 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     8101 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     8161 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     8221 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     8281 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     8341 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     8401 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     8461 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     8521 tgctgcttgc aaac
//
