LOCUS       sequence-340766-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRMCD77 encoding Wzz mutant G305A/G309A.
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: pRMCD112
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     75..92
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    107..246
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(172..191)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     332..354
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(348..539)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1052..1071
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1052..1070
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1102..1124
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2409..2427)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2423..2470
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2515..2533)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2601..2705
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2706..2774,2775..3566)
                     /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(2924..2943)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3737..4325
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4226..4245
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
       61 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
      121 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
      181 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
      241 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
      301 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
      361 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
      421 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
      481 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
      541 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
      601 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
      661 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
      721 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
      781 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
      841 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
      901 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
      961 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg
     1021 ctgcccgaga tcgatctcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa
     1081 cggtttccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgatgaga
     1141 gtagaaaata ataatgtttc tgggcaaaac catgacccgg aacagattga tttgattgat
     1201 ttactagtgc agttgtggcg tggcaagatg acaattatca tttccgtcat tgtggctatt
     1261 gccctggcta ttggttattt ggcagtagcg aaggagaaat ggacgtcaac agcaattatc
     1321 actcagcccg acgtggggca aattgctggc tataacaatg ccatgaatgt tatctatggt
     1381 caggctgcac cgaaagtatc ggatttgcag gagacgttaa ttggtcgctt cagttctgcc
     1441 ttctctgcat tagcagaaac gctggataat caggaagagc cagaaaaact taccatcgaa
     1501 ccttctgtta agaaccagca attaccattg actgtttctt atgttgggca aactgcagag
     1561 ggcgcacaaa tgaagttggc ccaatacatt cagcaagttg atgataaagt gaatcaagag
     1621 ctagaaaagg atctcaagga caacattgct ctgggacgga aaaacttgca ggactcttta
     1681 agaacccagg aagtggtcgc gcaggagcag aaagatctgc gtatccgtca gattcaggaa
     1741 gcgttgcagt atgcgaatca ggcgcaggtg acaaagccac aggttcagca gactgaagat
     1801 gtgacgcaag atacgttgtt ccttctaggg agcgaagcgc tggagtcgat gattaagcat
     1861 gaagcgactc gtccgttggt gttctcacca aactactatc agacacgtca aaacctgttg
     1921 gatattgaaa aattaaagtt tgatgatctt gatattcatg cttaccgcta tgtgatgaaa
     1981 ccgacgttac ctattcgtcg cgatagtccg aaaaaggcaa tcaccttgat tctggcagtg
     2041 cttctggccg gcatggttgc cgcggggatt gtgttggggc gtaacgctct gcgtaattac
     2101 aacgcgaagt aatattattg tgcatttaag agaaacgggc agggtggtga caccatgccc
     2161 gttttttttg ccggatgcga tgctggcgca tcttatccgg cctacgtgtg ttgagataat
     2221 gtgtaggcac gataagtttg cgcatcgggc aatggctccg ggtgtgacaa caacatcaca
     2281 cctgctcgga tccactagta acggccgcca gtgtgctgga attctgcaga tatccatcac
     2341 actggcggcc gctcgagcag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     2401 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     2461 gggttttttg ctgaaaggag gaactatatc cggataattc ttgaagacga aagggcctcg
     2521 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     2581 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2641 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2701 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2761 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2821 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2881 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2941 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3001 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3061 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3121 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3181 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3241 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3301 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3361 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3421 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3481 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3541 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3601 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3661 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3721 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3781 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3841 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3901 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3961 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4021 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4081 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4141 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4201 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4261 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4321 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4381 acatgttctt tcctgcgtta tccc
//
