pNZ8148乳酸菌分泌质粒

基本信息

启动子:	PnisA
复制子:	repA，repC
终止子:	T
质粒分类:	乳酸乳球菌表达质粒
质粒大小:	3167bp
原核抗性:	氯霉素Chl
筛选标记:	氯霉素Chl
克隆菌株:	NZ9000乳酸乳球菌
培养条件:	M17+0.5%葡萄糖/30度密封静置液体培养
表达宿主:	NZ9000乳酸乳球菌
培养条件:	M17+0.5%葡萄糖/30度密封静置液体培养
5'测序引物:	根据序列设计

质粒简介

          The pNZ8148 vector contains an origin of replication (ORI), the gene for the resistance to chloramphenicol, two genes for the replication proteins repA and repC, the nisin-inducible promoter (P nisA), and the transcription terminator (T). The gene, tagged with a Strep-tag II (STREP) followed by a stop codon (*), can be inserted between the necessary NcoI site and another endonuclease site from the multicloning site (MCS) such as PstI, SphI, KpnI, SpeI, XbaI, SacI and HindIII. The replicons of the vectors pNZ8008, pNZ8148, pNZ8149 and pNZ8150 are identical and came originally from the Lactococcus lactis plasmid pSH71. However, this replicon has a broad host range. Plasmids with this replicon can replicate in many Gram-positive bacteria, such as Lactobacillus plantarum and Streptococcus thermophilus. pNZ8148 – In this vector the nisA promoter is followed by an NcoI site for translational fusions at the ATG. It contains a terminator after the MCS. Sequence adaptation for cloning in NcoI can result in a change in the second amino acid of a protein (Mierau and Kleerebezem, 2005).

质粒图谱

质粒序列

LOCUS       Exported                3167 bp ds-DNA     circular SYN 29-AUG-2016
FEATURES             Location/Qualifiers
     source          1..3167
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    5..188
                     /note="Pnis"
     misc_feature    393..445
                     /note="Terminator"
     misc_feature    787..996
                     /note="repC"
     misc_feature    1264..1962
                     /note="repA"
     misc_feature    2421..3071
                     /note="Cm"
ORIGIN
        1 agatctagtc ttataactat actgacaata gaaacattaa caaatctaaa acagtcttaa
       61 ttctatcttg agaaagtatt ggtaataata ttattgtcga taacgcgagc ataataaacg
      121 gctctgatta aattctgaag tttgttagat acaatgattt cgttcgaagg aactacaaaa
      181 taaattataa ggaggcactc accatgggta ctgcaggcat gcggtaccac tagttctaga
      241 gagctcaagc tttctttgaa ccaaaattag aaaaccaagg cttgaaacgt tcaattgaaa
      301 tggcaattaa acaaattaca gcacgtgttg ctttgattga tagccaaaaa gcagcagttg
      361 ataaagcaat tactgatatt gctgaaaaat tgtaatttat aaataaaaat caccttttag
      421 aggtggtttt tttatttata aattattcgt ttgatttcgc tttcgataga acaatcaaat
      481 cgtttctgag acgttttagc gtttatttcg tttagttatc ggcataatcg ttaaaacagg
      541 cgttatcgta gcgtaaaagc ccttgagcgt agcgtggctt tgcagcgaag atgttgtctg
      601 ttagattatg aaagccgatg actgaatgaa ataataagcg cagcgtcctt ctatttcggt
      661 tggaggaggc tcaagggagt ttgagggaat gaaattccct catgggtttg attttaaaaa
      721 ttgcttgcaa ttttgccgag cggtagcgct ggaaaatttt tgaaaaaaat ttggaatttg
      781 gaaaaaaatg gggggaaagg aagcgaattt tgcttccgta ctacgacccc ccattaagtg
      841 ccgagtgcca atttttgtgc caaaaacgct ctatcccaac tggctcaagg gtttgagggg
      901 tttttcaatc gccaacgaat cgccaacgtt ttcgccaacg ttttttataa atctatattt
      961 aagtagcttt atttttgttt ttatgattac aaagtgatac actaatttta taaaattatt
     1021 tgattggagt tttttaaatg gtgatttcag aatcgaaaaa aagagttatg atttctctga
     1081 caaaagagca agataaaaaa ttaacagata tggcgaaaca aaaagatttt tcaaaatctg
     1141 cggttgcggc gttagctata gaagaatatg caagaaagga atcagaacaa aaaaaataag
     1201 cgaaagctcg cgtttttaga aggatacgag ttttcgctac ttgtttttga taaggtaatt
     1261 atatcatggc tattaaaaat actaaagcta gaaattttgg atttttatta tatcctgact
     1321 caattcctaa tgattggaaa gaaaaattag agagtttggg cgtatctatg gctgtcagtc
     1381 ctttacacga tatggacgaa aaaaaagata aagatacatg gaatagtagt gatgttatac
     1441 gaaatggaaa gcactataaa aaaccacact atcacgttat atatattgca cgaaatcctg
     1501 taacaataga aagcgttagg aacaagatta agcgaaaatt ggggaatagt tcagttgctc
     1561 atgttgagat acttgattat atcaaaggtt catatgaata tttgactcat gaatcaaagg
     1621 acgctattgc taagaataaa catatatacg acaaaaaaga tattttgaac attaatgatt
     1681 ttgatattga ccgctatata acacttgatg aaagccaaaa aagagaattg aagaatttac
     1741 ttttagatat agtggatgac tataatttgg taaatacaaa agatttaatg gcttttattc
     1801 gccttagggg agcggagttt ggaattttaa atacgaatga tgtaaaagat attgtttcaa
     1861 caaactctag cgcctttaga ttatggtttg agggcaatta tcagtgtgga tatagagcaa
     1921 gttatgcaaa ggttcttgat gctgaaacgg gggaaataaa atgacaaaca aagaaaaaga
     1981 gttatttgct gaaaatgagg aattaaaaaa agaaattaag gacttaaaag agcgtattga
     2041 aagatacaga gaaatggaag ttgaattaag tacaacaata gatttattga gaggagggat
     2101 tattgaataa ataaaagccc ccctgacgaa agtcgacggc aatagttacc cttattatca
     2161 agataagaaa gaaaaggatt tttcgctacg ctcaaatcct ttaaaaaaac acaaaagacc
     2221 acatttttta atgtggtctt ttattcttca actaaagcac ccattagttc aacaaacgaa
     2281 aattggataa agtgggatat ttttaaaata tatatttatg ttacagtaat attgactttt
     2341 aaaaaaggat tgattctaat gaagaaagca gacaagtaag cctcctaaat tcactttaga
     2401 taaaaattta ggaggcatat caaatgaact ttaataaaat tgatttagac aattggaaga
     2461 gaaaagagat atttaatcat tatttgaacc aacaaacgac ttttagtata accacagaaa
     2521 ttgatattag tgttttatac cgaaacataa aacaagaagg atataaattt taccctgcat
     2581 ttattttctt agtgacaagg gtgataaact caaatacagc ttttagaact ggttacaata
     2641 gcgacggaga gttaggttat tgggataagt tagagccact ttatacaatt tttgatggtg
     2701 tatctaaaac attctctggt atttggactc ctgtaaagaa tgacttcaaa gagttttatg
     2761 atttatacct ttctgatgta gagaaatata atggttcggg gaaattgttt cccaaaacac
     2821 ctatacctga aaatgctttt tctctttcta ttattccttg gacttcattt actgggttta
     2881 acttaaatat caataataat agtaattacc ttctacccat tattacagca ggaaaattca
     2941 ttaataaagg taattcaata tatttaccgc tatctttaca ggtacatcat tctgtttgtg
     3001 atggttatca tgctggattg tttatgaact ctattcagga attgtcagat aggcctaatg
     3061 actggctttt ataatatgag ataatgccga ctgtactttt tacagtcggt tttctaatgt
     3121 cactaacctg ccccgttagt tgaagaaggt ttttatatta cagctcc
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