LOCUS       sequence-263505-        4309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Medium 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 4309)
  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 4309)
  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_med
FEATURES             Location/Qualifiers
     source          1..4309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(145..162)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(279..824)
                     /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(359..378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(846..1655)
                     /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(1656..1747)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1814..1833
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gcgggacctc agcgctagcg gagtgtatac tggcttacta tgttggcact gatgagggtg
       61 tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat
      121 gtgatacagg atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact
      181 gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat
      241 acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct
      301 gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa
      361 agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg
      421 tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag
      481 ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga
      541 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc
      601 accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg
      661 gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc
      721 ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt
      781 cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta
      841 agcttttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
      901 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
      961 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     1021 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     1081 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     1141 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     1201 gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     1261 atcgagtgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     1321 ggatattctt ctaatacctg gaacgctgtt tttccgggga tcgcagtggt gagtaaccat
     1381 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gtggcataaa ttccgtcagc
     1441 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     1501 agaaacaact ctggcgcatc gggcttccca tacaagcgat agattgtcgc acctgattgc
     1561 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     1621 cgcggcctcg acgtttcccg ttgaatatgg ctcatattct tcctttttca atattattga
     1681 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     1741 aaacaaatag gggtcagtgt tacaaccaat taaccaattc tgaacattat cgcgagccca
     1801 tttatacctg aatatggctc ataacacccc ttgtttgcct ggcggcagta gcgcggtggt
     1861 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     1921 gactccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     1981 aagactgggc ctttcgcccg ggctaattag ggggtgtcgc ccttcgctga agtcgacgta
     2041 aaacgaatac cggggttatc ggtctgaatt gcgcaaagtt tacgtttaat tgtttttttt
     2101 aatagcggga ataaggggca gagaaaagag tatttcggcg actaacaaaa aatggctgtt
     2161 tttgaaaaaa attctaaagg ttgttttacg acagacgata acagggttga cggcgattga
     2221 gccgacgggt ggaaacccaa tacgtaatca acgacttgca atataggata acgaaccatg
     2281 gcacaagtca ttaataccaa cagcctctcg ctgatcactc aaaataatat caacaagaac
     2341 cagtctgcgc tgtcgagttc tatcgagcgt ctgtcttctg gcttgcgtat taacagcgcg
     2401 aaggatgacg cagcgggtca ggcgattgct aaccgtttca cctctaacat taaaggcctg
     2461 actcaggcgg cccgtaacgc caacgacggt atctccgttg cgcagaccac cgaaggcgcg
     2521 ctgtccgaaa tcaacaacaa cttacagcgt gtgcgtgaac tgacggtaca ggccactacc
     2581 ggtactaact ctgagtctga tctgtcttct atccaggacg aaattaaatc ccgtctggat
     2641 gaaattgacc gcgtatctgg tcagacccag ttcaacggcg tgaacgtgct ggcaaaaaat
     2701 ggctccatga aaatccaggt tggcgcaaat gataaccaga ctatcactat cgatctgaag
     2761 cagattgatg ctaaaactct tggccttgat ggttttagcg ttaaaaataa cgatacagtt
     2821 accactagtg ctccagtaac tgcttttggt gctaccacca caaacaatat taaacttact
     2881 ggaattaccc tttctacgga agcagccact gatactggcg gaactaaccc agcttcaatt
     2941 gagggtgttt atactgataa tggtaatgat tactatgcga aaatcaccgg tggtgataac
     3001 gatgggaagt attacgcagt aacagttgct aatgatggta cagtgacaat ggcgactgga
     3061 gcaacggcaa atgcaactgt aactgatgca aatactacta aagctacaac tatcacttca
     3121 ggcggtacac ctgttcagat tgataatact gcaggttccg caactgccaa ccttggtgct
     3181 gttagcttag taaaactgca ggattccaag ggtaatgata ccgatacata tgcgcttaaa
     3241 gatacaaatg gcaatcttta cgctgcggat gtgaatgaaa ctactggtgc tgtttctgtt
     3301 aaaactatta cctatactga ctcttccggt gccgccagtt ctccaaccgc ggtcaaactg
     3361 ggcggagatg atggcaaaac agaagtggtc gatattgatg gtaaaacata cgattctgcc
     3421 gatttaaatg gcggtaatct gcaaacaggt ttgactgctg gtggtgaggc tctgactgct
     3481 gttgcaaatg gtaaaaccac ggatccgctg aaagcgctgg acgatgctat cgcatctgta
     3541 gacaaattcc gttcttccct cggtgcggtg caaaaccgtc tggattccgc ggttaccaac
     3601 ctgaacaaca ccactaccaa cctgtctgaa gcgcagtccc gtattcagga cgccgactat
     3661 gcgaccgaag tgtccaatat gtcgaaagcg cagatcatcc agcaggccgg taactccgtg
     3721 ttggcaaaag ctaaccaggt accgcagcag gttctgtctc tgctgcaggg ttaagaattc
     3781 tgatcacact ggctaccttt gggtggcctt tttttatcgt caaaagggcg acacaaaagg
     3841 tattctaaat gcataataaa tactgataac atcttatagt ttgtattata ttttgtatta
     3901 tcgttgacat gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt
     3961 tattttctta attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa
     4021 tagatgaata gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa
     4081 gtgcgttgct tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac
     4141 aggcgccctt aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc
     4201 gccgaagcgg gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca
     4261 ggatgcctgg cagttcccta ctctcgccgc tgcgctcggt cgttcggct
//
