LOCUS       sequence-340777-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRMCD77 encoding Wzz mutant P292A.
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: pRMCD116
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     242..259
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    274..413
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(339..358)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     499..521
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(515..706)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1219..1238
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1219..1237
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1269..1291
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2576..2594)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2590..2637
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2682..2700)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2768..2872
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2873..2941,2942..3733)
                     /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(3091..3110)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3904..4404,1..88)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     join(4393..4404,1..8)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
       61 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      121 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      181 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      241 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
      301 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
      361 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
      421 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
      481 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
      541 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
      601 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
      661 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
      721 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
      781 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
      841 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
      901 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
      961 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     1021 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     1081 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     1141 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatcg
     1201 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
     1261 gaaataattt tgtttaactt taagaaggag atatacatat gatgagagta gaaaataata
     1321 atgtttctgg gctaaaccat gacccggaac agattgattt gattgattta ctagtgcagt
     1381 tgtggcgtgg caagatgaca attatcattt ccgtcattgt ggctattgcc ctggctattg
     1441 gttatttggc agtagcgaag gagaaatgga cgtcaacagc aattatcact cagcccgacg
     1501 tggggcaaat tgctggctat aacaatgcca tgaatgttat ctatggtcag gctgcaccga
     1561 aagtatcgga tttgcaggag acgttaattg gtcgcttcag ttctgccttc tctgcattag
     1621 cagaaacgct ggataatcag gaagagccag aaaaacttac catcgaacct tctgttaaga
     1681 accagcaatt accattgact gtttcttatg ttgggcaaac tgcagagggc gcacaaatga
     1741 agttggccca atacattcag caagttgatg ataaagtgaa tcaagagcta gaaaaggatc
     1801 tcaaggacaa cattgctctg ggacggaaaa acttgcagga ctctttaaga acccaggaag
     1861 tggtcgcgca ggagcagaaa gatctgcgta tccgtcagat tcaggaagcg ttgcagtatg
     1921 cgaatcaggc gcaggtgaca aagccacagg ttcagcagac tgaagatgtg acgcaagata
     1981 cgttgttcct tctagggagc gaagcgctgg agtcgatgat taagcatgaa gcgactcgtc
     2041 cgttggtgtt ctcaccaaac tactatcaga cacgtcaaaa cctgttggat attgaaaaat
     2101 taaagtttga tgatcttgat attcatgctt accgctatgt gatgaaaccg acgttaccta
     2161 ttcgtcgcga tagtgcgaaa aaggcaacca ccttgattct ggcagtgctt ctgggcggca
     2221 tggttggcgc ggggattgtg ttggggcgta acgctctgcg taattacaac gcgaagtaat
     2281 attattgtgc atttaagaga aacgggcagg gtggtgacac catgcccgtt ttttttgccg
     2341 gatgcgatgc tggcgcatct tatccggcct acgtgtgttg agataatgtg taggcacgat
     2401 aagtttgcgc atcgggcaat ggctccgggt gtgacaacaa catcacacct gctcggatcc
     2461 actagtaacg gccgccagtg tgctggaatt ctgcagatat ccatcacact ggcggccgct
     2521 cgagcagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2581 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     2641 aaaggaggaa ctatatccgg ataattcttg aagacgaaag ggcctcgtga tacgcctatt
     2701 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     2761 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2821 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2881 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2941 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3001 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3061 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     3121 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3181 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3241 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3301 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3361 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     3421 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3481 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3541 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3601 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3661 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3721 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3781 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3841 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3901 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3961 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4021 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     4081 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4141 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4201 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4261 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4321 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4381 ggagcttcca gggggaaacg cctg
//
