LOCUS       sequence-340774-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRMCD77 encoding Wzz mutant G305A/G311A.
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: pRMCD113
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             427..449
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1734..1752)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1748..1795
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1840..1858)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1926..2030
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2031..2099,2100..2891)
                     /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(2249..2268)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3062..3650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3551..3570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3804..3821
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3836..3975
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3901..3920)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4061..4083
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4077..4268)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
       61 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
      121 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
      181 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
      241 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
      301 aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgagatcgat
      361 ctcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt tccctctaga
      421 aataattttg tttaacttta agaaggagat atacatatga tgagagtaga aaataataat
      481 gtttctgggc aaaaccatga cccggaacag attgatttga ttgatttact agtgcagttg
      541 tggcgtggca agatgacaat tatcatttcc gtcattgtgg ctattgccct ggctattggt
      601 tatttggcag tagcgaagga gaaatggacg tcaacagcaa ttatcactca gcccgacgtg
      661 gggcaaattg ctggctataa caatgccatg aatgttatct atggtcaggc tgcaccgaaa
      721 gtatcggatt tgcaggagac gttaattggt cgcttcagtt ctgccttctc tgcattagca
      781 gaaacgctgg ataatcagga agagccagaa aaacttacca tcgaaccttc tgttaagaac
      841 cagcaattac cattgactgt ttcttatgtt gggcaaactg cagagggcgc acaaatgaag
      901 ttggcccaat acattcagca agttgatgat aaagtgaatc aagagctaga aaaggatctc
      961 aaggacaaca ttgctctggg acggaaaaac ttgcaggact ctttaagaac ccaggaagtg
     1021 gtcgcgcagg agcagaaaga tctgcgtatc cgtcagattc aggaagcgtt gcagtatgtg
     1081 aatcaggcgc aggtgacaaa gccacaggtt cagcagactg aagatgtgac gcaagatacg
     1141 ttgttccttc tagggagcga agcgctggag tcgatgatta agcatgaagc gactcgtccg
     1201 ttggtgttct caccaaacta ctatcagaca cgtcaaaacc tgttggatat tgaaaaatta
     1261 aagtttgatg atcttgatat tcatgcttac cgctatgtga tgaaaccgac gttacctatt
     1321 cgtcgcgata gtccgaaaaa ggcaatcacc ttgattctgg cagtgcttct ggccggcatg
     1381 gttggcgcgg cgattgtgtt ggggcgtaac gctctgcgta attacaacgc gaagtaatat
     1441 tattgtgcat ttaagagaaa cgggcagggt ggtgacacca tgcccgtttt ttttgccgga
     1501 tgcgatgctg gcgcatctta tccggcctac gtgtgttgag ataatgtgta ggcacgataa
     1561 gtttgcgcat cgagcaatgg ctccgggtgt gacaacaaca tcacacctgc tcggatccac
     1621 tagtaacggc cgccagtgtg ctggaattct gcagatatcc atcacactgg cggccgctcg
     1681 agcagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1741 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1801 aggaggaact atatccggat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     1861 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     1921 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1981 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2041 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2101 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2161 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2221 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     2281 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2341 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2401 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2461 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2521 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     2581 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2641 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2701 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2761 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2821 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2881 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2941 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3001 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3061 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3121 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3181 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3241 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3301 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3361 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3421 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3481 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3541 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3601 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3661 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3721 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3781 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3841 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     3901 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3961 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4021 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     4081 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     4141 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     4201 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     4261 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     4321 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     4381 cgttgtgagg gtaaacaact ggcg
//
