LOCUS       sequence-409424-        8831 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 8831)
  AUTHORS   Tang GQ, Paratkar S, Patel SS
  TITLE     Fluorescence mapping of the open complex of yeast mitochondrial RNA 
            polymerase.
  JOURNAL   J Biol Chem. 2009 Feb 27;284(9):5514-22. doi: 
            10.1074/jbc.M807880200. Epub 2008 Dec 30.
  PUBMED    19116203
REFERENCE   2  (bases 1 to 8831)
  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.M807880200"; journalName: "J Biol Chem"; date: 
            "2009-02-27- 27"; volume: "284"; issue: "9"; pages: "5514-22"
COMMENT     Sequence Label: RPO41
FEATURES             Location/Qualifiers
     source          1..8831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     939..958
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(978..1055)
                     /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(1133..1155)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1328..1347
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      1385..1813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1472..1491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1682..1703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1890..1907)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2061..2649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2141..2160)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2820..3611,3612..3680))
                     /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     3443..3462
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3681..3772)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(3791..3818)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3910..3996)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4149..4166
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(8349..8369)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(8391..8408)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8430..8452)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8441..8457
                     /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(8726..8831,1..977))
                     /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 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
       61 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
      121 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
      181 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
      241 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
      301 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
      361 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
      421 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
      481 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
      541 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
      601 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
      661 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
      721 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
      781 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
      841 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
      901 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
      961 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     1021 ataccgcgaa aggttttgca ccattcgatg gtgtcaacgt aaatgcatgc cgcttcgcct
     1081 tcgcgcgcga attgatctgc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     1141 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     1201 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     1261 gtagcgatag cggagtgtac cagtcacgta gcgatatcgc tacgtgactg gtactggctt
     1321 aactatgcgg catcagagca gattgtactg agagtgcacc atagcagcct gaatggcgaa
     1381 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1441 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1501 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1561 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1621 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1681 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1741 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1801 tttaacgcga attttaacaa aattatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     1861 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1921 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1981 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2041 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2101 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2161 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2221 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2281 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2341 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2401 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2461 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     2521 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2581 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2641 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2701 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2761 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2821 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2881 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2941 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3001 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3061 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3121 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3181 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3241 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3301 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3361 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3421 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3481 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3541 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3601 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3661 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3721 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aaagagtttg
     3781 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat gcctggcagt
     3841 ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt tcaaatccgc
     3901 tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     3961 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
     4021 catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgg
     4081 ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc agaccgcttc
     4141 tgcgttctga tttaatctgt atcaggctga aaatcttctc tcatccgcca aaacagccaa
     4201 gcttggtacc gcatgcctcg agactgcagg ctctagattc gaaagcggcc gcgactagtg
     4261 agctctcacg agaaaaaata ttgactgttt ctcaatactg taacatcaaa atcaccttta
     4321 ggtgggattt caggtaatct caacggtaaa agaacactca taccgctatt ttcatctcca
     4381 ccgttttcta gttcaagtgc gtccaagtca gtgatgtctt caaataatga tacagttgta
     4441 accatctttt caccaacatt tctatcttct attagagggc tgtttaataa ttcttgcctt
     4501 tttttctcaa aataaatttc atctgctaat gtagtacttc ttcctaattt tctggataaa
     4561 tctttacgga ttctaataat cttttgagcc aaatccgtag atctcttcaa ctttccaatc
     4621 ttgacgtaat tcttatacct ttggtcaaac tcctccttca atcttaaaac cagatctacc
     4681 tcatggagtt tgataaactg ttcccttaac accacattca tagtgtctat atcagatgca
     4741 tgcgtccagt aagaatcatg aactgatgcg aaatctaatc cttgctttcc acattctgct
     4801 gcacttagca acatatgtga agcatccagc gaatggatga agttcggtgg aagacctgcc
     4861 ttttgccttc ttgcattaac agggttgaca gcgaaaggat cgctaataaa gacagtttgc
     4921 aaattggtct caacttgctt tttgctttct tcacgatatg gctgaacaat tggtagtcct
     4981 agtggtgtcg tccatatgac ggatgacata aagtcgggtt tattaccgtt cttgaacgat
     5041 ttttcatcaa catccaacct gatagatttg gaaattcttt tcgcactttc acctaaccaa
     5101 tcctgaatta aatgcgcact atgaaataac tcgcggatag cactaaaaac atgtttggtt
     5161 aggtatttag aaaagtctaa gctttcttta cgatcatcaa atattgggct taattgttta
     5221 gcaatttgaa aagtagcccc gacgtaggta acaccataaa cgtttgtcat gacggtttgc
     5281 ttgacaacct ttctggtaat cttatccttc aaaattttgg cgttttcgtc tcctttttct
     5341 gctgcaattt ctaatctttt ttgcaccaac cttgcaacat gagcataaac atcttgtggt
     5401 ttatcgcttg gaaccagatt cacttgagta gccccttcaa cgtcaccacc caaggcagca
     5461 taatgttgca aaccattgca agtaccgtct tggtgaaccg gttgatgaga gataaactcc
     5521 tctggattat ccattttcat aacttcattc aattcaaagc atgtggccaa tgcttgccat
     5581 ggcttatcag cagtagtcca ccaacgatct cccgttaagg gattttccgc agaatccttg
     5641 atgtcctgca aatgagactc agtaaatgca acacgatcct ttaggggtaa cttatcaaat
     5701 ccgaacagat ttgacaaatg aatctttaac cacttgagac cagaaggtcc taatttttta
     5761 ccgtgccaaa aaatcaaaag gccacgactc atatcattac caagatgatt gaaatgtgga
     5821 gacaaaggat aagcacgtcc cctaaagtcc aaattatgtg gaaagtacag cttttcaccc
     5881 aaaaatgctc ttgcaatttc caatttgtag ttcgtatcac atctatttga tctatctgaa
     5941 gagaatttat tagcaattgt tttgacttga agtttccagg ctctcaaaat ggaagggtcg
     6001 gagttcttcg gaggagctgg tggtaaaacc atttcgtctt gtgcacctgg aatatctagg
     6061 aaaccttcac ctttattcca aacttgtgaa accacatcaa aaacttttct gttgacagtc
     6121 cacggtgttc tacccagaac attcaaacca tcataaacgc ggtctatatc accattatcg
     6181 gaagcagctt tcaaatatgc tacttgttcg ggtgagtctt ttgttcttaa aagggtagat
     6241 tgtgtataat gatatccacc tgatctccaa ttaacccagg gtttgggttc caccaacatt
     6301 ggtaataatt gtggttggac agaggcaatc aatctttcac cgttcaattg acgaatcaaa
     6361 gtcttgtgaa tcttcaagac acccaatttg gaaccattgt ggtactgata accatgcgca
     6421 aaagctggag cttcaccgtg aaccttagct tttgtcacag ggtcaacacc ttgcacagat
     6481 accttagcga cttgaatcaa cattgagatt aataccgaac cgattctggc cctaatactc
     6541 tgaggccaca aaattttcga ctgctcaatt tgagatgatc tgaatacaga tttagcattt
     6601 tgtaccagtt ttttgaattc tggtgatttc ttgttaacat ctctgaacgc ttgagattcg
     6661 cttttcaaaa cctgctcgga tctaaattcc atttcaatag ctttacccac tgatataacc
     6721 gctcttgcgg tcctcatacc ttcaataaca ccacccgtag aatttaactt taatagttcc
     6781 aaaatggtga tgacacacat tttaccaggg tcaatcaagg ttaagtatgg tccatatcca
     6841 agacggttcg tgtcaacttt attcaagcct ttcttatctg atagcttttc tgacaataat
     6901 gacctacaat gattaatttc ttccttcact agaggtaaca tttcattata ccatttccac
     6961 aacttaacgt ttaaattctt ttcaattgaa atgtcacctc tagcttttgc ctcttcaaaa
     7021 tcgtgcttcc aacgctctct tgcggcttcc gttgcacggt tttcaagtac cttttgtcta
     7081 tcttggttaa atatatttaa tgcgctttcg aatgccttct tttcctctaa tgttggcagt
     7141 gaattgtaaa tttcaaaaaa atttatagaa ttattattat cgtcgttttt cgtaggtttc
     7201 attttcaaaa cattatcatt ggcatcaaat ttgaatttag aaatttgctc tttctgttcc
     7261 ggcgttaatg ataatcccag caaagtatgc cggattactc tcattccaat agtgtctaca
     7321 gctataagct tttcagcgtc ttttgacaag acttctgttg aaccgtggaa tgaaacttcc
     7381 cgaagaggat ccaaggaagg tttttttata gaatcagagc ttgcatttga tgctttatgc
     7441 aattttgaat cgttctcagc ttccactttg ttcaaaccct cctcattttc aatattattt
     7501 acaggtgtag gagatagcgt aagtgattct aatattggtc ttacagaatt tggcagttgc
     7561 gatggtgtta taacctttaa gtcattcatt aaaatattca gatctgaaat ggtcaaaata
     7621 tctaggcagg agaaaatttc cctaatacca ttcacgctca ttttcaagta ggcagagatt
     7681 ataggcttca aaagcatcga gcttgtcgtt gaatggaagt tcaaagcgtg atgaatcatt
     7741 attgccaaag ttttgtcatt ataattaacg tctttaaaac ttgtctccaa atcatttgtt
     7801 aatttttcgt tcatttttaa aatagggaaa ttagggtcat ttttggaaaa tgaattcaag
     7861 tacatgttat aatcctcgat aaatcgctgc ttatgttcag gtaccaggta cagtgactcc
     7921 aatatagaga acgctctttt cattagattc gattgcaagc atgcttctaa cagagaccat
     7981 aattgcataa cgtctctagt tcttgtcatc gaggttactg ctgaggcatg cgatttattg
     8041 aaagggtcaa actctaattc tgaaggattt ttccattggt aggatatgct gctcttggcg
     8101 tctttcctat ttttgttgaa caaagggaaa tcatttgaac ttattatacc tgatgttgta
     8161 gatgaggttg gggacgagcc tgttacaagg gggtcttcag atattaatat tgtacttgtt
     8221 gagtccggag atggtttaaa cagcttagat ccctttgaag atattaaccg tctttgtaac
     8281 agggatgttt tcacgagcga tttataggcc ggtctcagca taggcctttg aattccggat
     8341 cccatggcgc cctgaaaata caggttttcg gtcgttggga tatcgtaatc gtgatggtga
     8401 tggtgatggt agtacgacat ggtctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     8461 cacagattat acggaccgga tgattaattg tcaacagctc atttcagaat atttgccaga
     8521 accgttatga tgtcggcgca aaaaacatta tccagaacgg gagtgcgcct tgagcgacac
     8581 gaattatgca gtgatttacg acctgcacag ccataccaca gcttccgatg gctgcctgac
     8641 gccagaagca ttggtgcacc gtgcagtcga tgataagctg tcaaaccaga tcaattcgcg
     8701 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     8761 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggtgcca
     8821 gggtggtttt t
//
