LOCUS       sequence-340785-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRMCD77 encoding Wzz mutant I35C/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: pRMCD122
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     195..214
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        195..213
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             245..267
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1552..1570)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1566..1613
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1658..1676)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1744..1848
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1849..1917,1918..2709)
                     /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(2067..2086)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2880..3468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3369..3388
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3622..3639
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3654..3793
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3719..3738)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3879..3901
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3895..4086)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
       61 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
      121 cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatcgatct
      181 cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc cctctagaaa
      241 taattttgtt taactttaag aaggagatat acatatgatg agagtagaaa ataataatgt
      301 ttctgggcaa aaccatgacc cggaacagat tgatttgatt gatttactag tgcagttgtg
      361 gcgtggcaag acgacaattt gcatttccgt cattgcggct attgccctgg ctattggtta
      421 tttggcagta gcgaaggaga aatggacgtc aacagcaatt atcactcagc ccgacgtggg
      481 gcaaattgct ggctataaca atgccatgaa tgttatctat ggtcaggctg caccgaaagt
      541 atcggatttg caggagacgt taattggtcg cttcagttct gccttctctg cattagcaga
      601 aacgctggat aatcaggaag agccagaaaa acttaccatc gaaccttctg ttaagaacca
      661 gcaattacca ttgactgttt cttatgttgg gcaaactgca gagggcgcac aaatgaagtt
      721 ggcccaatac attcagcaag ttgatgataa agtgaatcaa gagctagaaa aggatctcaa
      781 ggacaacatt gctctgggac ggaaaaactt gcaggactct ttaagaaccc aggaagtggt
      841 cgcgcaggag cagatagatc tgcgtatccg tcagattcag gaagcgttgc agtatgcgaa
      901 tcaggcgcag gtgacaaagc cacaggttca gcagactgaa gatgtgacgc aagatacgtt
      961 gttccttcta gggagcgaag cgctggagtc gatgattaag catgaagcga ctcgtccgtt
     1021 ggtgttctca ccaaactact atcagacacg tcaaaacctg ttggatattg aaaaattaaa
     1081 gtttgatgat cttgatattc atgcttaccg ctatgtgatg aaaccgacgt tacctattcg
     1141 tcgcgatagt ccgaaaaagg caatcacctt gattctggca gtgcttctgg gcggcatggt
     1201 tggcgcgggg attgtgttgg ggcgtaacgc tctgcgtaat tacaacgcga agtaatatta
     1261 ttgtgcattt aagagaaacg ggcagggtgg tgacaccatg cccgtttttt ttgccggatg
     1321 cgatgctggc gcatcttatc cggcctacgt gtgttgagat aatgtgtagg cacgataagt
     1381 ttgcgcatcg ggcaatggct ccgggtgtga caacaacatc acacctgctc ggatccacta
     1441 gtaacggccg ccagtgtgct ggaattctgc agatatccat cacactggcg gccgctcgag
     1501 cagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     1561 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     1621 gaggaactat atccggataa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     1681 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     1741 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1801 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     1861 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     1921 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1981 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2041 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     2101 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2161 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2221 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2281 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2341 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     2401 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2461 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2521 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     2581 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     2641 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     2701 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     2761 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2821 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2881 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2941 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3001 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3061 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3121 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3181 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3241 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3301 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3361 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3421 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3481 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3541 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3601 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     3661 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     3721 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     3781 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3841 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3901 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     3961 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     4021 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     4081 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     4141 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     4201 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     4261 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     4321 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     4381 cgaaacacgg aaaccgaaga ccat
//
