LOCUS       sequence-271732-        5661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low 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 5661)
  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 5661)
  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_low
FEATURES             Location/Qualifiers
     source          1..5661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        172..290
                     /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      2379..2910
                     /label=oriV
                     /note="incP origin of replication"
     CDS             3303..4451
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     terminator      complement(4514..4541)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(4557..4628)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4641..5450)
                     /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(5451..5542)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5609..5628
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc
       61 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
      121 gggcctttcg cccgggctaa ttagggggtg tcgcccttcg ctgaagtcga ccaccacaat
      181 tcagcaaatt gtgaacatca tcacgttcat ctttccctgg ttgccaatgg cccattttcc
      241 tgtcagtaac gagaaggtcg cgtattcagg cgctttttag actggtcgta atgaaattca
      301 gcaggatcac accatggcac aagtcattaa taccaacagc ctctcgctga tcactcaaaa
      361 taatatcaac aagaaccagt ctgcgctgtc gagttctatc gagcgtctgt cttctggctt
      421 gcgtattaac agcgcgaagg atgacgcagc gggtcaggcg attgctaacc gtttcacctc
      481 taacattaaa ggcctgactc aggcggcccg taacgccaac gacggtatct ccgttgcgca
      541 gaccaccgaa ggcgcgctgt ccgaaatcaa caacaactta cagcgtgtgc gtgaactgac
      601 ggtacaggcc actaccggta ctaactctga gtctgatctg tcttctatcc aggacgaaat
      661 taaatcccgt ctggatgaaa ttgaccgcgt atctggtcag acccagttca acggcgtgaa
      721 cgtgctggca aaaaatggct ccatgaaaat ccaggttggc gcaaatgata accagactat
      781 cactatcgat ctgaagcaga ttgatgctaa aactcttggc cttgatggtt ttagcgttaa
      841 aaataacgat acagttacca ctagtgctcc agtaactgct tttggtgcta ccaccacaaa
      901 caatattaaa cttactggaa ttaccctttc tacggaagca gccactgata ctggcggaac
      961 taacccagct tcaattgagg gtgtttatac tgataatggt aatgattact atgcgaaaat
     1021 caccggtggt gataacgatg ggaagtatta cgcagtaaca gttgctaatg atggtacagt
     1081 gacaatggcg actggagcaa cggcaaatgc aactgtaact gatgcaaata ctactaaagc
     1141 tacaactatc acttcaggcg gtacacctgt tcagattgat aatactgcag gttccgcaac
     1201 tgccaacctt ggtgctgtta gcttagtaaa actgcaggat tccaagggta atgataccga
     1261 tacatatgcg cttaaagata caaatggcaa tctttacgct gcggatgtga atgaaactac
     1321 tggtgctgtt tctgttaaaa ctattaccta tactgactct tccggtgccg ccagttctcc
     1381 aaccgcggtc aaactgggcg gagatgatgg caaaacagaa gtggtcgata ttgatggtaa
     1441 aacatacgat tctgccgatt taaatggcgg taatctgcaa acaggtttga ctgctggtgg
     1501 tgaggctctg actgctgttg caaatggtaa aaccacggat ccgctgaaag cgctggacga
     1561 tgctatcgca tctgtagaca aattccgttc ttccctcggt gcggtgcaaa accgtctgga
     1621 ttccgcggtt accaacctga acaacaccac taccaacctg tctgaagcgc agtcccgtat
     1681 tcaggacgcc gactatgcga ccgaagtgtc caatatgtcg aaagcgcaga tcatccagca
     1741 ggccggtaac tccgtgttgg caaaagctaa ccaggtaccg cagcaggttc tgtctctgct
     1801 gcagggttaa gaattctgat cacactggct acctttgggt ggcctttttt tatcgtcaaa
     1861 agggcgacac aaaaggtatt ctaaatgcat aataaatact gataacatct tatagtttgt
     1921 attatatttt gtattatcgt tgacatgtat aattttgata tcaaaaactg attttccctt
     1981 tattattttc gagatttatt ttcttaattc tctttaacaa actagaaata ttgtatatac
     2041 aaaaaatcat aaataataga tgaatagttt aattataggt gttcatcaat cgaaaaagca
     2101 acgtatctta tttaaagtgc gttgcttttt tctcatttat aaggttaaat aattctcata
     2161 tatcaagcaa agtgacaggc gcccttaaat attctgacaa atgctctttc cctaaactcc
     2221 ccccataaaa aaacccgccg aagcgggttt ttacgttatt tgcggattaa cgattactcg
     2281 ttatcagaac cgcccaggat gcctggcagt tccctactct cgccgctgcg ctcggtcgtt
     2341 cggctgcggg acctcagcgc tagcgcagag ccatgtaggg ccgccggcgt tgtggatacc
     2401 tcgcggaaaa cttggccctc actgacagat gaggggcgga cgttgacact tgaggggccg
     2461 actcacccgg cgcggcgttg acagatgagg ggcaggctcg atttcggccg gcgacgtgga
     2521 gctggccagc ctcgcaaatc ggcgaaaacg cctgatttta cgcgagtttc ccacagatga
     2581 tgtggacaag cctggggata agtgccctgc ggtattgaca cttgaggggc gcgactactg
     2641 acagatgagg ggcgcgatcc ttgacacttg aggggcagag tgctgacaga tgaggggcgc
     2701 acctattgac atttgagggg ctgtccacag gcagaaaatc cagcatttgc aagggtttcc
     2761 gcccgttttt cggccaccgc taacctgtct tttaacctgc ttttaaacca atatttataa
     2821 accttgtttt taaccagggc tgcgccctgt gcgcgtgacc gcgcacgccg aaggggggtg
     2881 cccccccttc tcgaaccctc ccggcccgct aacgcgggcc tcccatcccc ccaggggctg
     2941 cgcccctcgg ccgcgaacgg cctcacccca aaaatggcag ccacgtagaa agccagtccg
     3001 cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac
     3061 gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg
     3121 gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt
     3181 gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg
     3241 ggatcaagat cgacggatcg atccggggaa ttaattccgg ggcaatcccg caaggagggt
     3301 gaatgaatcg gacgtttgac cggaaggcat acaggcaaga actgatcgac gcggggtttt
     3361 ccgccgagga tgccgaaacc atcgcaagcc gcaccgtcat gcgtgcgccc cgcgaaacct
     3421 tccagtccgt cggctcgata gtccagcaag ctacggccaa gatcgagcgc gacagcgtgc
     3481 aactggctcc ccctgccctg cccgcgccat cggccgccgt ggagcgttcg cgtcgtctcg
     3541 aacaggaggc ggcaggtttg gcgaagtcga tgaccatcga cacgcgagga actatgacga
     3601 ccaagaagcg aaaaaccgcc ggcgaggacc tggcaaaaca ggtcagcgaa gccaagcagg
     3661 ccgcgttgct gaaacacacg aagcagcaga tcaaggaaat gcagctttcc ttgttcgata
     3721 ttgcgccgtg gccggacacg atgcgagcga tgccaaacga cacggcccgc tctgccctgt
     3781 tcaccacgcg caacaagaaa atcccgcgcg aggcgctgca aaacaaggtc attttccacg
     3841 tcaacaagga cgtgaagatc acctacaccg gcgtcgagct gcgggccgac gatgacgaac
     3901 tggtgtggca gcaggtgttg gagtacgcga agcgcacccc tatcggcgag ccgatcacct
     3961 tcacgttcta cgagctttgc caggacctgg gctggtcgat caatggccgg tattacacga
     4021 aggccgagga atgcctgtcg cgcctacagg cgacggcgat gggcttcacg tccgaccgcg
     4081 ttgggcacct ggaatcggtg tcgctgctgc accgcttccg cgtcctggac cgtggcaaga
     4141 aaacgtcccg ttgccaggtc ctgatcgacg aggaaatcgt cgtgctgttt gctggcgacc
     4201 actacacgaa attcatatgg gagaagtacc gcaagctgtc gccgacggcc cgacggatgt
     4261 tcgactattt cagctcgcac cgggagccgt acccgctcaa gctggaaacc ttccgcctca
     4321 tgtgcggatc ggattccacc cgcgtgaaga agtggcgcga gcaggtcggc gaagcctgcg
     4381 aagagttgcg aggcagcggc ctggtggaac acgcctgggt caatgatgac ctggtgcatt
     4441 gcaaacgcta gggccttgtg gggtcagttc cggctggggg ttcagcagcc acctgcatcg
     4501 ccctaggtat aaacgcagaa aggcccaccc gaaggtgagc cagtgtgact ctagtagaga
     4561 gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt
     4621 tttatttgat gcctggaagc ttagaaaaac tcatcgagca tcaaatgaaa ttgcaattta
     4681 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     4741 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     4801 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     4861 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     4921 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     4981 ccgttattca ttcgtgattg cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga
     5041 caattacaaa caggaatcga gtgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     5101 ttttcacctg aatcaggata ttcttctaat acctggaacg ctgtttttcc ggggatcgca
     5161 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagtggc
     5221 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     5281 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa gcgatagatt
     5341 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     5401 atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat attcttcctt
     5461 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5521 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac
     5581 attatcgcga gcccatttat acctgaatat ggctcataac accccttgtt tgcctggcgg
     5641 cagtagcgcg gtggtcccac c
//
