LOCUS       sequence-340786-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRMCD77 encoding Wzz mutant G305A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4404)
  AUTHORS   Daniels C, Morona R
  TITLE     Analysis of Shigella flexneri wzz (Rol) function by mutagenesis and 
            cross-linking: wzz is able to oligomerize.
  JOURNAL   Mol Microbiol. 1999 Oct;34(1):181-94. doi: 
            10.1046/j.1365-2958.1999.01591.x.
  PUBMED    10540296
REFERENCE   2  (bases 1 to 4404)
  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: 
            "10.1046/j.1365-2958.1999.01591"; journalName: "Mol Microbiol"; 
            date: "1999-10"; volume: "34"; issue: "1"; pages: "181-94"
COMMENT     Sequence Label: pRMCD125
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     427..446
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        427..445
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             477..499
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1784..1802)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1798..1845
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1890..1908)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1976..2080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2081..2149,2150..2941)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2299..2318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3112..3700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3601..3620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3854..3871
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3886..4025
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3951..3970)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4111..4133
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4127..4318)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
       61 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
      121 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
      181 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
      241 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
      301 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
      361 gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgagatcgat ctcgatcccg
      421 cgaaattaat acgactcact atagggagac cacaacggtt tccctctaga aataattttg
      481 tttaacttta agaaggagat atacatatga tgagagtaga aaataataat gtttctgggc
      541 aaaaccatga cccggaacag attgatttga ttgatttact agtgcagttg tggcgtggca
      601 agatgacaat tatcatttcc gtcattgtgg ctattgccct ggctattggt tatttggcag
      661 tagcgaagga gaaatggacg tcaacagcaa ttatcactca gcccgacgtg gggcaaattg
      721 ctggctataa caatgccatg aatgttatct atggtcaggc tgcaccgaaa gtatcggatt
      781 tgcaggagac gttaattggt cgcttcagtt ctgccttctc tgcattagca gaaacgctgg
      841 ataatcagga agagccagaa aaacttacca tcgaaccttc tgttaagaac cagcaattac
      901 cattgactgt ttcttatgtt gggcaaactg cagagggcgc acaaatgaag ttggcccaat
      961 acattcagca agttgatgat aaagtgaatc aagagctaga aaaggatctc aaggacaaca
     1021 ttgctctggg acggaaaaac ttgcaggact ctttaagaac ccaggaagtg gtcgcgcagg
     1081 agcagaaaga tctgcgtatc cgtcagattc aggaagcgtt gcagtatgcg aatcaggcgc
     1141 aggtgacaaa gccacaggtt cagcagactg aagatgtgac gcaagatacg ttgttccttc
     1201 tagggagcga agcgctggag tcgatgatta agcatgaggc gactcgtccg ttggtgttct
     1261 caccaaacta ctatcagaca cgtcaaaacc tgttggatat tgaaaaatta aagtttgatg
     1321 atcttgatat tcatgcttac cgctatgtga tgaaaccgac gttacctatt cgtcgcgata
     1381 gtccgaaaaa ggcaatcacc ttgattctgg cagtgcttct ggccggcatg gttggcgcgg
     1441 ggattgtgtt ggggcgtaac gctctgcgta attacaacgc gaagtaatat tattgtgcat
     1501 ttaagagaaa cgggcagggt ggtgacacca tgcccgtttt ttttgccgga tgcgatgctg
     1561 gcgcatctta tccggcctac gtgtgttgag ataatgtgta ggcacgataa gtttgcgcat
     1621 cgggcaatgg ctccgggtgt gacaacaaca tcacacctgc tcggatccac tagtaacggc
     1681 cgccagtgtg ctggaattct gcagatatcc atcacactgg cggccgctcg agcagatccg
     1741 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     1801 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     1861 atatccggat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     1921 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     1981 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2041 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2101 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2161 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2221 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2281 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     2341 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     2401 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     2461 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     2521 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     2581 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     2641 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     2701 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     2761 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     2821 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     2881 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     2941 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3001 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3061 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3121 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3181 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3241 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3301 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3361 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3421 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3481 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3541 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3601 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3661 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3721 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3781 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3841 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     3901 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     3961 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     4021 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4081 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4141 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     4201 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     4261 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     4321 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     4381 ggatgctcac gatacgggtt actg
//
