LOCUS       sequence-340781-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRMCD77 encoding Wzz mutant I35C.
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: pRMCD121
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     140..159
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        140..158
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             190..212
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1497..1515)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1511..1558
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1603..1621)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1689..1793
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1794..1862,1863..2654)
                     /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(2012..2031)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2825..3413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3314..3333
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3567..3584
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3599..3738
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3664..3683)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3824..3846
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3840..4031)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
       61 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatc
      121 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
      181 agaaataatt ttgtttaact ttaagaagga gatatacata tgatgagagt agaaaataat
      241 aatgtttctg ggcaaaacca tgacccggaa cagattgatt tgattgattt actagtgcag
      301 ttgtggcgtg gcaagatgac aatttgcatt tccgtcattg tggctattgc cctggctatt
      361 ggttatttgg cagtagcgaa ggagaaatgg acgtcaacag caattatcac tcagcccgac
      421 gtggggcaaa ttgctggcta taacaatgcc atgaatgtta tctatggtca ggctgcaccg
      481 aaagtatcgg atttgcagga gacgttaatt ggtcgcttca gttctgcctt ctctgcatta
      541 gcagaaacgc tggataatca ggaagagcca gaaaaactta ccatcgaacc ttctgttaag
      601 aaccagcaat taccattgac tgtttcttat gttgggcaaa ctgcagaggg cgcacaaatg
      661 aagttggccc aatacattca gcaagttgat gataaagtga atcaagagct agaaaaggat
      721 ctcaaggaca acattgctct gggacggaaa aacttgcagg actctttaag aacccaggaa
      781 gtggtcgcgc aggagcagaa agatctgcgt atccgtcaga ttcaggaagc gttgcagtat
      841 gcgaatcagg cgcaggtgac aaagccacag gttcagcaga ctgaagatgt gacgcaagat
      901 acgttgttcc ttctagggag cgaagcgctg gagtcgatga ttaagcatga agcgactcgt
      961 ccgttggtgt tctcaccaaa ctactatcag acacgtcaaa acctgttgga tattgaaaaa
     1021 ttaaagtttg atgatcttga cattcatgct taccgctatg tgatgaaacc gacgttacct
     1081 attcgtcgcg atagtccgaa aaaggcaatc accttgattc tggcagtgct tctgggcggc
     1141 atggttggcg cggggattgt gttggggcgt aacgctctgc gtaattacaa cgcgaagtaa
     1201 tattattgtg catttaagag aaacgggcag ggtggtgaca ccatgcccgt tttttttgcc
     1261 ggatgcgatg ctggcgcatc ttatccggcc tacgtgcgtt gagataatgt gtaggcacga
     1321 taagtttgcg catcgggcaa tggctccggg tgtgacaaca acatcacacc tgctcggatc
     1381 cactagtaac ggccgccagt gtgctggaat tctgcagata tccatcacac tggcggccgc
     1441 tcgagcagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     1501 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     1561 gaaaggagga actatatccg gataattctt gaagacgaaa gggcctcgtg atacgcctat
     1621 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     1681 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1741 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1801 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1861 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1921 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1981 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     2041 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2101 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2161 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2221 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2281 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     2341 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2401 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2461 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     2521 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2581 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2641 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2701 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2761 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2821 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2881 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2941 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     3001 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3061 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3121 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3181 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3241 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3301 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3361 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3421 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3481 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3541 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3601 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     3661 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     3721 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3781 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3841 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     3901 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     3961 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     4021 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     4081 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     4141 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     4201 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     4261 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     4321 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     4381 cagcagcagt cgcttcacgt tcgc
//
