LOCUS       sequence-271730-        3978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High 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 3978)
  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 3978)
  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_hi
FEATURES             Location/Qualifiers
     source          1..3978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        105..223
                     /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)"
     rep_origin      complement(2298..2886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2378..2397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2891..3700)
                     /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(3701..3792)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3859..3878
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
       61 tcgcccgggc taattagggg gtgtcgccct tcgctgaagt cgaccaccac aattcagcaa
      121 attgtgaaca tcatcacgtt catctttccc tggttgccaa tggcccattt tcctgtcagt
      181 aacgagaagg tcgcgtattc aggcgctttt tagactggtc gtaatgaaat tcagcaggat
      241 cacaccatgg cacaagtcat taataccaac agcctctcgc tgatcactca aaataatatc
      301 aacaagaacc agtctgcgct gtcgagttct atcgagcgtc tgtcttctgg cttgcgtatt
      361 aacagcgcga aggatgacgc agcgggtcag gcgattgcta accgtttcac ctctaacatt
      421 aaaggcctga ctcaggcggc ccgtaacgcc aacgacggta tctccgttgc gcagaccacc
      481 gaaggcgcgc tgtccgaaat caacaacaac ttacagcgtg tgcgtgaact gacggtacag
      541 gccactaccg gtactaactc tgagtctgat ctgtcttcta tccaggacga aattaaatcc
      601 cgtctggatg aaattgaccg cgtatctggt cagacccagt tcaacggcgt gaacgtgctg
      661 gcaaaaaatg gctccatgaa aatccaggtt ggcgcaaatg ataaccagac tatcactatc
      721 gatctgaagc agattgatgc taaaactctt ggccttgatg gttttagcgt taaaaataac
      781 gatacagtta ccactagtgc tccagtaact gcttttggtg ctaccaccac aaacaatatt
      841 aaacttactg gaattaccct ttctacggaa gcagccactg atactggcgg aactaaccca
      901 gcttcaattg agggtgttta tactgataat ggtaatgatt actatgcgaa aatcaccggt
      961 ggtgataacg atgggaagta ttacgcagta acagttgcta atgatggtac agtgacaatg
     1021 gcgactggag caacggcaaa tgcaactgta actgatgcaa atactactaa agctacaact
     1081 atcacttcag gcggtacacc tgttcagatt gataatactg caggttccgc aactgccaac
     1141 cttggtgctg ttagcttagt aaaactgcag gattccaagg gtaatgatac cgatacatat
     1201 gcgcttaaag atacaaatgg caatctttac gctgcggatg tgaatgaaac tactggtgct
     1261 gtttctgtta aaactattac ctatactgac tcttccggtg ccgccagttc tccaaccgcg
     1321 gtcaaactgg gcggagatga tggcaaaaca gaagtggtcg atattgatgg taaaacatac
     1381 gattctgccg atttaaatgg cggtaatctg caaacaggtt tgactgctgg tggtgaggct
     1441 ctgactgctg ttgcaaatgg taaaaccacg gatccgctga aagcgctgga cgatgctatc
     1501 gcatctgtag acaaattccg ttcttccctc ggtgcggtgc aaaaccgtct ggattccgcg
     1561 gttaccaacc tgaacaacac cactaccaac ctgtctgaag cgcagtcccg tattcaggac
     1621 gccgactatg cgaccgaagt gtccaatatg tcgaaagcgc agatcatcca gcaggccggt
     1681 aactccgtgt tggcaaaagc taaccaggta ccgcagcagg ttctgtctct gctgcagggt
     1741 taagaattct gatcacactg gctacctttg ggtggccttt ttttatcgtc aaaagggcga
     1801 cacaaaaggt attctaaatg cataataaat actgataaca tcttatagtt tgtattatat
     1861 tttgtattat cgttgacatg tataattttg atatcaaaaa ctgattttcc ctttattatt
     1921 ttcgagattt attttcttaa ttctctttaa caaactagaa atattgtata tacaaaaaat
     1981 cataaataat agatgaatag tttaattata ggtgttcatc aatcgaaaaa gcaacgtatc
     2041 ttatttaaag tgcgttgctt ttttctcatt tataaggtta aataattctc atatatcaag
     2101 caaagtgaca ggcgccctta aatattctga caaatgctct ttccctaaac tccccccata
     2161 aaaaaacccg ccgaagcggg tttttacgtt atttgcggat taacgattac tcgttatcag
     2221 aaccgcccag gatgcctggc agttccctac tctcgccgct gcgctcggtc gttcggctgc
     2281 gggacctcag cgctagcttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2341 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2401 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2461 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2521 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2581 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2641 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2701 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2761 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2821 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2881 tctcaagctt ttagaaaaac tcatcgagca tcaaatgaaa ttgcaattta ttcatatcag
     2941 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     3001 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     3061 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     3121 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     3181 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     3241 ttcgtgattg cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga caattacaaa
     3301 caggaatcga gtgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     3361 aatcaggata ttcttctaat acctggaacg ctgtttttcc ggggatcgca gtggtgagta
     3421 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagtggc ataaattccg
     3481 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     3541 gtttcagaaa caactctggc gcatcgggct tcccatacaa gcgatagatt gtcgcacctg
     3601 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     3661 ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat attcttcctt tttcaatatt
     3721 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3781 aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac attatcgcga
     3841 gcccatttat acctgaatat ggctcataac accccttgtt tgcctggcgg cagtagcgcg
     3901 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt
     3961 gtggggactc cccatgcg
//
