LOCUS       sequence-340756-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRMCD77 encoding Wzz mutant K267N.
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: pRMCD108
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     149..168
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        149..167
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             199..221
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1506..1524)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1520..1567
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1612..1630)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1698..1802
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1803..1871,1872..2663)
                     /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(2021..2040)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2834..3422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3323..3342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3576..3593
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3608..3747
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3673..3692)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3833..3855
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3849..4040)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
       61 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg
      121 cccgagatcg atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg
      181 tttccctcta gaaataattt tgtttaactt taagaaggag atatacatat gatgagagta
      241 gaaaataata atgtttctgg gcaaaaccat gacccggaac agattgattt gattgattta
      301 ctagtgcagt tgtggcgtgg caagatgaca attatcattt ccgtcattgt ggctattgcc
      361 ctggctattg gttatttggc agtagcgaag gagaaatgga cgtcaacagc aattatcact
      421 cagcccgacg tggggcaaat tgctggctat aacaatgcca tgaatgttat ctatggtcag
      481 gctgcaccga aagtatcgga tttgcaggag acgttaattg gtcgcttcag ttctgccttc
      541 tctgcattag cagaaacgct ggataatcag gaagagccag aaaaacttac catcgaacct
      601 tctgttaaga accagcaatt accattgact gtttcttatg ttgggcaaac tgcagagggc
      661 gcacaaatga agttggccca atacattcag caagttgatg ataaagtgaa tcaagagcta
      721 gaaaaggatc tcaaggacaa cattgctctg ggacggaaaa acttgcagga ctctttaaga
      781 acccaggaag tggtcgcgca ggagcagaaa gatctgcgta tccgtcagat tcaggaagcg
      841 ttgcagtatg cgaatcaggc gcaggtgaca aagccacagg ttcagcagac tgaagatgtg
      901 acgcaagata cgttgttcct tctagggagc gaagcgctgg agtcgatgat taagcatgaa
      961 gcgactcgtc cgttggtgtt ctcaccaaac tactatcaga cacgtcaaaa tctgttggat
     1021 attgaaaact taaagtttga tgatcttgat attcatgctt accgctatgt gatgaaaccg
     1081 acgttaccta ttcgtcgcga tagtccgaaa aaggcaatca ccttgattct ggcagtgctt
     1141 ctgggcggca tggttggcgc ggggattgtg ttggggcgta acgctctgcg taattacaac
     1201 gcgaagtaat attattgtgc atttaagaga aacgggcagg gtggtgacac catgcccgtt
     1261 ttttttgccg gatgcgatgc tggcgcatct tatccggcct acgtgtgttg agataatgtg
     1321 taggcacgat aagtttgcgc atcgggcaat ggctccgggt gtgacaacaa catcacacct
     1381 gctcggatcc actagtaacg gccgccagtg tgctggaatt ctgcagatat ccatcacact
     1441 ggcggccgct cgagcagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1501 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1561 ttttttgctg aaaggaggaa ctatatccgg ataattcttg aagacgaaag ggcctcgtga
     1621 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     1681 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1741 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1801 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1861 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1921 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1981 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2041 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2101 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2161 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2221 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2281 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2341 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2401 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2461 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2521 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2581 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2641 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2701 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2761 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2821 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2881 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2941 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3001 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3061 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3121 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3181 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3241 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3301 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3361 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3421 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3481 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3541 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3601 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     3661 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     3721 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     3781 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3841 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     3901 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     3961 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     4021 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     4081 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     4141 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     4201 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     4261 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     4321 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     4381 gacgttttgc agcagcagtc gctt
//
