LOCUS       sequence-271735-        5759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy number plasmid for expression of E. coli flagellin from its
            natural promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5759)
  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 5759)
  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_WT_low
FEATURES             Location/Qualifiers
     source          1..5759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1524..2055
                     /label=oriV
                     /note="incP origin of replication"
     CDS             2448..3596
                     /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(3659..3686)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(3702..3773)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3786..4595)
                     /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(4596..4687)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4754..4773
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 taccactagt gctccagtaa ctgcttttgg tgctaccacc acaaacaata ttaaacttac
       61 tggaattacc ctttctacgg aagcagccac tgatactggc ggaactaacc cagcttcaat
      121 tgagggtgtt tatactgata atggtaatga ttactatgcg aaaatcaccg gtggtgataa
      181 cgatgggaag tattacgcag taacagttgc taatgatggt acagtgacaa tggcgactgg
      241 agcaacggca aatgcaactg taactgatgc aaatactact aaagctacaa ctatcacttc
      301 aggcggtaca cctgttcaga ttgataatac tgcaggttcc gcaactgcca accttggtgc
      361 tgttagctta gtaaaactgc aggattccaa gggtaatgat accgatacat atgcgcttaa
      421 agatacaaat ggcaatcttt acgctgcgga tgtgaatgaa actactggtg ctgtttctgt
      481 taaaactatt acctatactg actcttccgg tgccgccagt tctccaaccg cggtcaaact
      541 gggcggagat gatggcaaaa cagaagtggt cgatattgat ggtaaaacat acgattctgc
      601 cgatttaaat ggcggtaatc tgcaaacagg tttgactgct ggtggtgagg ctctgactgc
      661 tgttgcaaat ggtaaaacca cggatccgct gaaagcgctg gacgatgcta tcgcatctgt
      721 agacaaattc cgttcttccc tcggtgcggt gcaaaaccgt ctggattccg cggttaccaa
      781 cctgaacaac accactacca acctgtctga agcgcagtcc cgtattcagg acgccgacta
      841 tgcgaccgaa gtgtccaata tgtcgaaagc gcagatcatc cagcaggccg gtaactccgt
      901 gttggcaaaa gctaaccagg taccgcagca ggttctgtct ctgctgcagg gttaagaatt
      961 ctgatcacac tggctacctt tgggtggcct ttttttatcg tcaaaagggc gacacaaaag
     1021 gtattctaaa tgcataataa atactgataa catcttatag tttgtattat attttgtatt
     1081 atcgttgaca tgtataattt tgatatcaaa aactgatttt ccctttatta ttttcgagat
     1141 ttattttctt aattctcttt aacaaactag aaatattgta tatacaaaaa atcataaata
     1201 atagatgaat agtttaatta taggtgttca tcaatcgaaa aagcaacgta tcttatttaa
     1261 agtgcgttgc ttttttctca tttataaggt taaataattc tcatatatca agcaaagtga
     1321 caggcgccct taaatattct gacaaatgct ctttccctaa actcccccca taaaaaaacc
     1381 cgccgaagcg ggtttttacg ttatttgcgg attaacgatt actcgttatc agaaccgccc
     1441 aggatgcctg gcagttccct actctcgccg ctgcgctcgg tcgttcggct gcgggacctc
     1501 agcgctagcg cagagccatg tagggccgcc ggcgttgtgg atacctcgcg gaaaacttgg
     1561 ccctcactga cagatgaggg gcggacgttg acacttgagg ggccgactca cccggcgcgg
     1621 cgttgacaga tgaggggcag gctcgatttc ggccggcgac gtggagctgg ccagcctcgc
     1681 aaatcggcga aaacgcctga ttttacgcga gtttcccaca gatgatgtgg acaagcctgg
     1741 ggataagtgc cctgcggtat tgacacttga ggggcgcgac tactgacaga tgaggggcgc
     1801 gatccttgac acttgagggg cagagtgctg acagatgagg ggcgcaccta ttgacatttg
     1861 aggggctgtc cacaggcaga aaatccagca tttgcaaggg tttccgcccg tttttcggcc
     1921 accgctaacc tgtcttttaa cctgctttta aaccaatatt tataaacctt gtttttaacc
     1981 agggctgcgc cctgtgcgcg tgaccgcgca cgccgaaggg gggtgccccc ccttctcgaa
     2041 ccctcccggc ccgctaacgc gggcctccca tccccccagg ggctgcgccc ctcggccgcg
     2101 aacggcctca ccccaaaaat ggcagccacg tagaaagcca gtccgcagaa acggtgctga
     2161 ccccggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga
     2221 aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca
     2281 gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa
     2341 gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc aagatcgacg
     2401 gatcgatccg gggaattaat tccggggcaa tcccgcaagg agggtgaatg aatcggacgt
     2461 ttgaccggaa ggcatacagg caagaactga tcgacgcggg gttttccgcc gaggatgccg
     2521 aaaccatcgc aagccgcacc gtcatgcgtg cgccccgcga aaccttccag tccgtcggct
     2581 cgatagtcca gcaagctacg gccaagatcg agcgcgacag cgtgcaactg gctccccctg
     2641 ccctgcccgc gccatcggcc gccgtggagc gttcgcgtcg tctcgaacag gaggcggcag
     2701 gtttggcgaa gtcgatgacc atcgacacgc gaggaactat gacgaccaag aagcgaaaaa
     2761 ccgccggcga ggacctggca aaacaggtca gcgaagccaa gcaggccgcg ttgctgaaac
     2821 acacgaagca gcagatcaag gaaatgcagc tttccttgtt cgatattgcg ccgtggccgg
     2881 acacgatgcg agcgatgcca aacgacacgg cccgctctgc cctgttcacc acgcgcaaca
     2941 agaaaatccc gcgcgaggcg ctgcaaaaca aggtcatttt ccacgtcaac aaggacgtga
     3001 agatcaccta caccggcgtc gagctgcggg ccgacgatga cgaactggtg tggcagcagg
     3061 tgttggagta cgcgaagcgc acccctatcg gcgagccgat caccttcacg ttctacgagc
     3121 tttgccagga cctgggctgg tcgatcaatg gccggtatta cacgaaggcc gaggaatgcc
     3181 tgtcgcgcct acaggcgacg gcgatgggct tcacgtccga ccgcgttggg cacctggaat
     3241 cggtgtcgct gctgcaccgc ttccgcgtcc tggaccgtgg caagaaaacg tcccgttgcc
     3301 aggtcctgat cgacgaggaa atcgtcgtgc tgtttgctgg cgaccactac acgaaattca
     3361 tatgggagaa gtaccgcaag ctgtcgccga cggcccgacg gatgttcgac tatttcagct
     3421 cgcaccggga gccgtacccg ctcaagctgg aaaccttccg cctcatgtgc ggatcggatt
     3481 ccacccgcgt gaagaagtgg cgcgagcagg tcggcgaagc ctgcgaagag ttgcgaggca
     3541 gcggcctggt ggaacacgcc tgggtcaatg atgacctggt gcattgcaaa cgctagggcc
     3601 ttgtggggtc agttccggct gggggttcag cagccacctg catcgcccta ggtataaacg
     3661 cagaaaggcc cacccgaagg tgagccagtg tgactctagt agagagcgtt caccgacaaa
     3721 caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg
     3781 gaagcttaga aaaactcatc gagcatcaaa tgaaattgca atttattcat atcaggatta
     3841 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     3901 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     3961 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     4021 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     4081 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     4141 gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     4201 atcgagtgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     4261 ggatattctt ctaatacctg gaacgctgtt tttccgggga tcgcagtggt gagtaaccat
     4321 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gtggcataaa ttccgtcagc
     4381 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     4441 agaaacaact ctggcgcatc gggcttccca tacaagcgat agattgtcgc acctgattgc
     4501 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     4561 cgcggcctcg acgtttcccg ttgaatatgg ctcatattct tcctttttca atattattga
     4621 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4681 aaacaaatag gggtcagtgt tacaaccaat taaccaattc tgaacattat cgcgagccca
     4741 tttatacctg aatatggctc ataacacccc ttgtttgcct ggcggcagta gcgcggtggt
     4801 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     4861 gactccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     4921 aagactgggc ctttcgcccg ggctaattag ggggtgtcgc ccttcgctga agtcgacgta
     4981 aaacgaatac cggggttatc ggtctgaatt gcgcaaagtt tacgtttaat tgtttttttt
     5041 aatagcggga ataaggggca gagaaaagag tatttcggcg actaacaaaa aatggctgtt
     5101 tttgaaaaaa attctaaagg ttgttttacg acagacgata acagggttga cggcgattga
     5161 gccgacgggt ggaaacccaa tacgtaatca acgacttgca atataggata acgaaccatg
     5221 gcacaagtca ttaataccaa cagcctctcg ctgatcactc aaaataatat caacaagaac
     5281 cagtctgcgc tgtcgagttc tatcgagcgt ctgtcttctg gcttgcgtat taacagcgcg
     5341 aaggatgacg cagcgggtca ggcgattgct aaccgtttca cctctaacat taaaggcctg
     5401 actcaggcgg cccgtaacgc caacgacggt atctccgttg cgcagaccac cgaaggcgcg
     5461 ctgtccgaaa tcaacaacaa cttacagcgt gtgcgtgaac tgacggtaca ggccactacc
     5521 ggtactaact ctgagtctga tctgtcttct atccaggacg aaattaaatc ccgtctggat
     5581 gaaattgacc gcgtatctgg tcagacccag ttcaacggcg tgaacgtgct ggcaaaaaat
     5641 ggctccatga aaatccaggt tggcgcaaat gataaccaga ctatcactat cgatctgaag
     5701 cagattgatg ctaaaactct tggccttgat ggttttagcg ttaaaaataa cgatacagt
//
