LOCUS       sequence-340779-        4404 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pRMCD77 encoding Wzz mutant K31A.
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 Apr 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: pRMCD119
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     212..229
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    244..383
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(309..328)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     469..491
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(485..676)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1189..1208
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1189..1207
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1239..1261
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2546..2564)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2560..2607
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2652..2670)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2738..2842
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2843..2911,2912..3703)
                     /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(3061..3080)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3874..4404,1..58)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4363..4382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
       61 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      121 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      181 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      241 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
      301 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
      361 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
      421 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
      481 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
      541 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
      601 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
      661 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
      721 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
      781 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
      841 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
      901 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
      961 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     1021 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     1081 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     1141 gtggccagga cccaacgctg cccgagatcg atctcgatcc cgcgaaatta atacgactca
     1201 ctatagggag accacaacgg tttccctcta gaaataattt tgtttaactt taagaaggag
     1261 atatacatat gatgagagta gaaaataata atgtttctgg gcaaaaccat gacccggaac
     1321 agattgattt gattgattta ctagtgcagt tgtggcgtgg cgcgatgaca attatcattt
     1381 ccgtcattgt ggctattgcc ctggctattg gttatttggc agtagcgaag gagaaatgga
     1441 cgtcaacagc aattatcact cagcccgacg tggggcaaat tgctggctat aacaatgcca
     1501 tgaatgttat ctatggtcag gctgcaccga aagtatcgga tttgcaggag acgttaattg
     1561 gtcgcttcag ttctgccttc tctgtattag cagaaacgct ggataatcag gaagagccag
     1621 aaaaacttac catcgaacct tctgttaaga accagcaatt accattgact gtttcttatg
     1681 ttgggcaaac tgcagagggc gcacaaatga agttggccca atacattcag caagttgatg
     1741 ataaagtgaa tcaagagcta gaaaaggatc tcaaggacaa cattgctctg ggacggaaaa
     1801 acttgcagga ctctttaaga acccaggaag tggtcgcgca ggagcagaaa gatctgtgta
     1861 tccgtcagat tcaggaagcg ttgcagtatg cgaatcaggc gcaggtgaca aagccacagg
     1921 ttcagcagac tgaagatgtg acgcaagata cgttgttcct tctagggagc gaagcgctgg
     1981 agtcgatgat taagcatgaa gcgactcgtc cgttggtgtt ctcaccaaac tactatcaga
     2041 cacgtcaaaa cctgttggat attgaaaaat taaagtttga tgatcttgat attcatgctt
     2101 accgctatgt gatgaaaccg acgttaccta ttcgtcgcga tagtccgaaa aaggcaatca
     2161 ccttgattct ggcagtgctt ctgggcggca tggttggcgc ggggattgtg ttggggcgta
     2221 acgctctgcg taattacaac gcgaagtaat attattgtgc atttaagaga aacgggcagg
     2281 gtggtgacac catgcccgtt ttttttgccg gatgcgatgc tggcgcatct tatccggcct
     2341 acgtgtgttg ggataatgtg taggcacgat aagtttgcgc atcgggcaat ggctccgggt
     2401 gtgacaacaa catcacacct gctcggatcc actagtaacg gccgccagtg tgctggaatt
     2461 ctgcagatat ccatcacact ggcggccgct cgagcagatc cggctgctaa caaagcccga
     2521 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     2581 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg ataattcttg
     2641 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     2701 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2761 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2821 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2881 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2941 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3001 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3061 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     3121 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3181 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3241 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3301 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3361 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     3421 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3481 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3541 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3601 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3661 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3721 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3781 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3841 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3901 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3961 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4021 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4081 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4141 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4201 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4261 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4321 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4381 ttatagtcct gtcgggtttc gcca
//
