LOCUS       sequence-271728-        4209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Medium copy number plasmid for expression of E. coli flagellin from 
            a rhamnose-inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4209)
  AUTHORS   Thomson NM, Pallen MJ
  TITLE     Restoration of wild-type motility to flagellin-knockout Escherichia 
            coli by varying promoter, copy number and induction strength in 
            plasmid-based expression of flagellin.
  JOURNAL   Curr Res Biotechnol. 2020 Nov;2:45-52. doi: 
            10.1016/j.crbiot.2020.03.001.
  PUBMED    33381753
REFERENCE   2  (bases 1 to 4209)
  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"; journalName: "Curr Res 
            Biotechnol."; date: "2020-11"; volume: "2:45-52. doi"; pages: " 
            10.1016/j.crbiot.2020.03.001"
COMMENT     Sequence Label: pJ211_rha_WT_med
FEATURES             Location/Qualifiers
     source          1..4209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        155..273
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     primer_bind     complement(2473..2490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2607..3152)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2687..2706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3172..3981)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(3982..4073)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4140..4159
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cagaagtgaa acgccgtagc gccgatggta gtgtggggac tccccatgcg agagtaggga
       61 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgcccgggc
      121 taattagggg gtgtcgccct tcgctgaagt cgaccaccac aattcagcaa attgtgaaca
      181 tcatcacgtt catctttccc tggttgccaa tggcccattt tcctgtcagt aacgagaagg
      241 tcgcgtattc aggcgctttt tagactggtc gtaatgaaat tcagcaggat cacaccatgg
      301 cacaagtcat taataccaac agcctctcgc tgatcactca aaataatatc aacaagaacc
      361 agtctgcgct gtcgagttct atcgagcgtc tgtcttctgg cttgcgtatt aacagcgcga
      421 aggatgacgc agcgggtcag gcgattgcta accgtttcac ctctaacatt aaaggcctga
      481 ctcaggcggc ccgtaacgcc aacgacggta tctccgttgc gcagaccacc gaaggcgcgc
      541 tgtccgaaat caacaacaac ttacagcgtg tgcgtgaact gacggtacag gccactaccg
      601 gtactaactc tgagtctgat ctgtcttcta tccaggacga aattaaatcc cgtctggatg
      661 aaattgaccg cgtatctggt cagacccagt tcaacggcgt gaacgtgctg gcaaaaaatg
      721 gctccatgaa aatccaggtt ggcgcaaatg ataaccagac tatcactatc gatctgaagc
      781 agattgatgc taaaactctt ggccttgatg gttttagcgt taaaaataac gatacagtta
      841 ccactagtgc tccagtaact gcttttggtg ctaccaccac aaacaatatt aaacttactg
      901 gaattaccct ttctacggaa gcagccactg atactggcgg aactaaccca gcttcaattg
      961 agggtgttta tactgataat ggtaatgatt actatgcgaa aatcaccggt ggtgataacg
     1021 atgggaagta ttacgcagta acagttgcta atgatggtac agtgacaatg gcgactggag
     1081 caacggcaaa tgcaactgta actgatgcaa atactactaa agctacaact atcacttcag
     1141 gcggtacacc tgttcagatt gataatactg caggttccgc aactgccaac cttggtgctg
     1201 ttagcttagt aaaactgcag gattccaagg gtaatgatac cgatacatat gcgcttaaag
     1261 atacaaatgg caatctttac gctgcggatg tgaatgaaac tactggtgct gtttctgtta
     1321 aaactattac ctatactgac tcttccggtg ccgccagttc tccaaccgcg gtcaaactgg
     1381 gcggagatga tggcaaaaca gaagtggtcg atattgatgg taaaacatac gattctgccg
     1441 atttaaatgg cggtaatctg caaacaggtt tgactgctgg tggtgaggct ctgactgctg
     1501 ttgcaaatgg taaaaccacg gatccgctga aagcgctgga cgatgctatc gcatctgtag
     1561 acaaattccg ttcttccctc ggtgcggtgc aaaaccgtct ggattccgcg gttaccaacc
     1621 tgaacaacac cactaccaac ctgtctgaag cgcagtcccg tattcaggac gccgactatg
     1681 cgaccgaagt gtccaatatg tcgaaagcgc agatcatcca gcaggccggt aactccgtgt
     1741 tggcaaaagc taaccaggta ccgcagcagg ttctgtctct gctgcagggt taagaattct
     1801 gatcacactg gctacctttg ggtggccttt ttttatcgtc aaaagggcga cacaaaaggt
     1861 attctaaatg cataataaat actgataaca tcttatagtt tgtattatat tttgtattat
     1921 cgttgacatg tataattttg atatcaaaaa ctgattttcc ctttattatt ttcgagattt
     1981 attttcttaa ttctctttaa caaactagaa atattgtata tacaaaaaat cataaataat
     2041 agatgaatag tttaattata ggtgttcatc aatcgaaaaa gcaacgtatc ttatttaaag
     2101 tgcgttgctt ttttctcatt tataaggtta aataattctc atatatcaag caaagtgaca
     2161 ggcgccctta aatattctga caaatgctct ttccctaaac tccccccata aaaaaacccg
     2221 ccgaagcggg tttttacgtt atttgcggat taacgattac tcgttatcag aaccgcccag
     2281 gatgcctggc agttccctac tctcgccgct gcgctcggtc gttcggctgc gggacctcag
     2341 cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc
     2401 ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat
     2461 atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga
     2521 aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga
     2581 agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac
     2641 gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg
     2701 tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc
     2761 attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc
     2821 agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt
     2881 atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc
     2941 agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa
     3001 ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt
     3061 tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc
     3121 aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaag cttagaaaaa
     3181 ctcatcgagc atcaaatgaa attgcaattt attcatatca ggattatcaa taccatattt
     3241 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     3301 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     3361 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     3421 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     3481 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     3541 gaggcgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg agtgcaaccg
     3601 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     3661 tacctggaac gctgtttttc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     3721 acggataaaa tgcttgatgg tcggaagtgg cataaattcc gtcagccagt ttagtctgac
     3781 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     3841 cgcatcgggc ttcccataca agcgatagat tgtcgcacct gattgcccga cattatcgcg
     3901 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgacgt
     3961 ttcccgttga atatggctca tattcttcct ttttcaatat tattgaagca tttatcaggg
     4021 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4081 cagtgttaca accaattaac caattctgaa cattatcgcg agcccattta tacctgaata
     4141 tggctcataa caccccttgt ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     4201 tgccgaact
//
