LOCUS       sequence-254586-        4217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli rpoz gene cloned between NcoI and HindIII sites of the first
            MCS of this Duet plasmid. Omega protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4217)
  AUTHORS   Murakami KS
  TITLE     X-ray Crystal Structure of Escherichia coli RNA Polymerase sigma70 
            Holoenzyme.
  JOURNAL   J Biol Chem. 2013 Mar 29;288(13):9126-34. doi: 
            10.1074/jbc.M112.430900. Epub 2013 Feb 6.
  PUBMED    23389035
REFERENCE   2  (bases 1 to 4217)
  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.M112.430900"; journalName: "J Biol Chem"; date: 
            "2013-03-29- 29"; volume: "288"; issue: "13"; pages: "9126-34"
COMMENT     Sequence Label: pACYCDuet-1_Ec_rpoZ
FEATURES             Location/Qualifiers
     source          1..4217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             88..363
                     /codon_start=1
                     /gene="E. coli rpoZ"
                     /product="DNA-directed RNA polymerase subunit omega"
                     /label=RNAP omega
                     /note="promotes RNA polymerase assembly"
                     /translation="MARVTVQDAVEKIGNRFDLVLVAARRARQMQVGGKDPLVPEENDK
                     TTVIALREIEEGLINNQILDVRERQEQQEQEAAELQAVTAIAEGRR"
     promoter        438..456
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    457..481
                     /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             590..634
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(672..690)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      686..733
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(954..1613)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1528..1547
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1614..1716)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2108..2125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2242..2787)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2322..2341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3012..4094)
                     /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     4056..4075
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4095..4172)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg gcacgcgtaa ctgttcagga cgctgtagag
      121 aaaattggta accgttttga cctggtactg gtcgccgcgc gtcgcgctcg tcagatgcag
      181 gtaggcggaa aggatccgct ggtaccggaa gaaaacgata aaaccactgt aatcgcgctg
      241 cgcgaaatcg aagaaggtct gatcaacaac cagatcctcg acgttcgcga acgccaggaa
      301 cagcaagagc aggaagccgc tgaattacaa gccgttaccg ctattgctga aggtcgtcgt
      361 taataaaagc ttgcggccgc ataatgctta agtcgaacag aaagtaatcg tattgtacac
      421 ggccgcataa tcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
      481 cccatcttag tatattagtt aagtataaga aggagatata catatggcag atctcaattg
      541 gatatcggcc ggccacgcga tcgctgacgt cggtaccctc gagtctggta aagaaaccgc
      601 tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt aattaaccta
      661 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
      721 gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac tgcttccggt
      781 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
      841 accgacgacc gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt
      901 attcaggcgt agcaaccagg cgtttaaggg caccaataac tgccttaaaa aaattacgcc
      961 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     1021 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     1081 gtataatatt tgcccatagt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     1141 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
     1201 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     1261 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     1321 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     1381 ttcattgcca tacggaactc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     1441 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     1501 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     1561 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     1621 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     1681 ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg
     1741 gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct
     1801 tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt
     1861 tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca gctgtccctc
     1921 ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac atcagcgcta
     1981 gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga agtgcttcat
     2041 gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt
     2101 ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg
     2161 cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga
     2221 gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat
     2281 ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2341 ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
     2401 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     2461 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     2521 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     2581 ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa
     2641 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     2701 gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag
     2761 attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc
     2821 tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat
     2881 aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     2941 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     3001 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3061 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     3121 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     3181 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     3241 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca
     3301 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     3361 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     3421 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     3481 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     3541 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     3601 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     3661 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     3721 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     3781 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     3841 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     3901 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     3961 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     4021 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     4081 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     4141 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga
     4201 ctcctgcatt aggaaat
//
