pKK223-3

pKK223-3

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pKK223-3,Plasmid pKK223-3,pKK223-3 vector

pKK223-3 Plasmid Information

Bacterial Resistance:Ampicillin
Growth Strain:DH5

pKK223-3 Plasmid Description

pKK223-3 (1) contains the strong tac promoter upstream from the multiple cloning site and the strong rrnB ribosomal terminator downstream for control of protein expression (2-4). Induction: tac promoter is inducible with 1-5 mM IPTG. Promoter is very strong; even uninduced cells may show a low level of expression. Expression: Genes containing a ribosome-binding site and ATG can be inserted into any unique site in the MCS for expression. The ribosomebinding site on the plasmid can be utilised to express inserts if the ATG start codon of the insert is within 5-13 bp from the provided ribosomebindingsite (5, 6). Transcription terminators: rrnB transcription terminators stabilize the plasmid by inhibiting read-through transcription initiated from the tac promoter in the parent plasmid. Host(s): lac Iq strains; E. coli JM105 is recommended. Selectable marker(s): Plasmid confers resistance to 100 μg/ml ampicillin. Amplification: Recommended. Control Regions: Expression control region: tac promoter: -10: 50-44; -35: 72-67; Ribosome binding site: 11. MCS: 4552-4587. rrnB operon region: 5S rRNA region: 4471-4352; rrnB T1 terminator: 4348-4305; rrnB T2 terminator: 4173-4146. b-lactamase gene region: Promoter: -10: 4082-4077; -35: 4105-4100; Start codon (ATG): 4035; Stop codon (TAA): 3178. Plasmid replication region: Site of replication initiation: 2417-2415; Region necessary for replication: 3111-2415.

pKK223-3 Plasmid Sequence
LOCUS       Exported                4584 bp ds-DNA     circular SYN 18-涔濇湀-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4584)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 2016-09-18 from SnapGene Viewer 2.8.1
            
FEATURES             Location/Qualifiers
     source          1..4584
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    20..36
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     misc_feature    complement(44..72)
                     /note="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             1796..1987
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2089..2229
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2415..3003)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        complement(4034..4125)
                     /gene="bla"
                     /note="AmpR promoter"
     terminator      4144..4171
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      4263..4349
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacattata cgagccgatg
       61 attaattgtc aacagctcat ttcagaatat ttgccagaac cgttatgatg tcggcgcaaa
      121 aaacattatc cagaacggga gtgcgccttg agcgacacga attatgcagt gatttacgac
      181 ctgcacagcc ataccacagc ttccgatggc tgcctgacgc cagaagcatt ggtgcaccgt
      241 gcagtcgata agcccggatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
      301 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
      361 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
      421 gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc
      481 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgaccga
      541 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
      601 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
      661 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc
      721 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
      781 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg
      841 tcttgctggc gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt
      901 ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc
      961 atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcactggac
     1021 cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga
     1081 ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc
     1141 gggccacctc gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag
     1201 aattggagcc aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa
     1261 catatccatc gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg
     1321 gtcctggcca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg
     1381 gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg
     1441 ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc
     1501 gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc
     1561 aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg
     1621 accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca
     1681 acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg
     1741 tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac
     1801 caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct
     1861 tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca
     1921 cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa
     1981 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     2041 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     2101 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     2161 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     2221 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2281 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2341 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2401 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2461 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2521 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2581 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     2641 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2701 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2761 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc