LOCUS       sequence-340788-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRMCD77 encoding Wzz mutant M32T.
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: pRMCD127
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     399..418
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        399..417
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             449..471
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1756..1774)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1770..1817
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1862..1880)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1948..2052
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2053..2121,2122..2913)
                     /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(2271..2290)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3084..3672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3573..3592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3826..3843
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3858..3997
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3923..3942)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4083..4105
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4099..4290)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
       61 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
      121 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
      181 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
      241 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
      301 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga
      361 cccaacgctg cccgagatcg atctcgatcc cgcgaaatta atacgactca ctatagggag
      421 accacaacgg tttccctcta gaaataattt tgtttaactt taagaaggag atatacatat
      481 gatgagagta gaaaataata atgtttctgg gcaaaaccat gacccggaac agattgattt
      541 gattgattta ctagtgcagt tgtggcgtgg caagacgaca attatcattt ccgtcattgt
      601 ggctattgcc ctggctattg gttatttggc agtagcgaag gagaaatgga cgtcaacagc
      661 aattatcact cagcccgacg tggggcaaat tgctggctat aacaatgcca tgaatgttat
      721 ctatggtcag gctgcaccga aagtatcgga tttgcaggag acgttaattg gtcgcttcag
      781 ttctgccttc tctgcattag cagaaacgct ggataatcag gaagagccag aaaaacttac
      841 catcgaacct tctgttaaga accagcaatt accattgact gtttcttatg ttgggcaaac
      901 tgcagagggc gcacaaatga agttggccca atacattcag caagttgatg ataaagtgaa
      961 tcaagagcta gaaaaggatc tcaaggacaa cattgctctg ggacggaaaa acttgcagga
     1021 ctctttaaga acccaggaag tggtcgcgca ggagcagaaa gatctgcgta tccgtcagat
     1081 tcaggaagcg ttgcagtatg cgaatcaggc gcaggtgaca aagccacagg ttcagcagac
     1141 tgaagatgtg acgcaagata cgttgttcct tctagggagc gaagcgctgg agtcgatgat
     1201 taagcatgaa gcgactcgtc cgttggtgtt ctcaccaaac tactatcaga cacgtcaaaa
     1261 cctgttggat attgaaaaat taaagtttga tgatcttgat attcatgctt accactatgt
     1321 gatgaaaccg acgttaccta ttcgtcgcga tagtccgaaa aaggcaatca ccttgattct
     1381 ggcagtgctt ctgggcggca tggttggcgc ggggattgtg ttggggcgta acgctctgcg
     1441 taattacaac gcgaagtaat attattgtgc atttaagaga aacgggcagg gtggtgacac
     1501 catgcccgtt ttttttgccg gatgcgatgc tggcgcatct tatccggcct acgtgtgttg
     1561 agataatgtg taggcacgat aagtttgcgc atcgggcaat ggctccgggt gtgacaacaa
     1621 catcacacct gctcggatcc actagtaacg gccgccagtg tgctggaatt ctgcagatat
     1681 ccatcacact ggcggccgct cgagcagatc cggctgctaa caaagcccga aaggaagctg
     1741 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1801 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg ataattcttg aagacgaaag
     1861 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     1921 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1981 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2041 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2101 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2161 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2221 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2281 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2341 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2401 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2461 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2521 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2581 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2641 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     2701 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2761 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2821 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2881 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     2941 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3001 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3061 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3121 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3181 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3241 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3301 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3361 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3421 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3481 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3541 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3601 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3661 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3721 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3781 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3841 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3901 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     3961 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     4021 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4081 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4141 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4201 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4261 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     4321 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     4381 tgaacatgcc cggttactgg aacg
//
