LOCUS       sequence-340757-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRMCD77 encoding Wzz mutant P283A.
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: pRMCD109
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     104..123
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        104..122
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             154..176
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1461..1479)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1475..1522
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1567..1585)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1653..1757
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1758..1826,1827..2618)
                     /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(1976..1995)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2789..3377
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3278..3297
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3531..3548
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3563..3702
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3628..3647)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3788..3810
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3804..3995)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
       61 caggacccaa cgctgcccga gatcgatctc gatcccgcga aattaatacg actcactata
      121 gggagaccac aacggtttcc ctctagaaat aattttgttt aactttaaga aggagatata
      181 catatgatga gagtagaaaa taataatgtt tctgggcaaa accatgaccc ggaacagatt
      241 gatttgattg atttactagt gcagttgtgg cgtggcaaga tgacaattat catttccgtc
      301 attgtggcta ttgccctggc tattggttat ttggcagtag cgaaggagaa atggacgtca
      361 acagcaatta tcactcagcc cgacgtgggg caaattgctg gctataacaa tgccatgaat
      421 gttatctatg gtcaggctgc accgaaagta tcggatttgc aggagacgtt aattggtcgc
      481 ttcagttctg ccttctctgc attagcagaa acgctggata atcaggaaga gccagaaaaa
      541 cttaccatcg aaccttctgt taagaaccag caattaccat tgactgtttc ttatgttggg
      601 caaactgccg agggcgcaca aatgaagttg gcccaataca ttcagcaagt tgatgataaa
      661 gtgaatcaag agctagaaaa ggatctcaag gacaacattg ctctgggacg gaaaaacttg
      721 caggactctt taagaaccca ggaagtggtc gcgcaggagc agaaagatct gcgtatccgt
      781 cagattcagg aagcgttgca gtatgcgaat caggcgcagg tgacaaagcc acaggttcag
      841 cagactgaag atgtgacgca agatacgttg ttccttctag ggagcgaagc gctggagtcg
      901 atgattaagc atgaagcgac tcgtccgttg gtgttctcac caaactacta tcagacacgt
      961 caaaacctgt tggatattga aaaattaaag tttgatgatc ttgatattca tgcttaccgc
     1021 tatgtgatga aagcgacgtt acctattcgt cgcgatagtc cgaaaaaggc aatcaccttg
     1081 attctggcag tgcttctggg cggcatggtt ggcgcgggga ttgtgttggg gcgtaacgct
     1141 ctgcgtaatt acaacgcgaa gtaatattat tgtgcattta agagaaacgg gcagggtggt
     1201 gacaccatgc ccgttttttt tgccggatgc gatgctggcg catcttatcc ggcctacgtg
     1261 tgttgagata atgtgtaggc acgataagtt tgcgcatcgg gcaatggctc cgggtgtgac
     1321 aacaacatca cacctgctcg gatccactag taacggccgc cagtgtgctg gaattctgca
     1381 gatatccatc acactggcgg ccgctcgagc agatccggct gctaacaaag cccgaaagga
     1441 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1501 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggataat tcttgaagac
     1561 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     1621 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1681 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1741 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1801 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1861 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1921 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1981 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     2041 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2101 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2161 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2221 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2281 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2341 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2401 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2461 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2521 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2581 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2641 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2701 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2761 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2821 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2881 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     2941 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3001 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3061 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3121 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3181 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3241 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3301 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3361 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3421 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3481 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3541 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3601 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3661 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     3721 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3781 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     3841 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     3901 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     3961 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     4021 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     4081 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     4141 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     4201 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     4261 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     4321 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     4381 gattcattct gctaaccagt aagg
//
