| 质粒名称: | pJAK112 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 167280 |
| 存储实验室: | Robert Fagan |
|
载体骨架基本信息 |
|
| 载体名称: | pMTLSC-7215 |
| 载体出品公司: | Nigel P Minton |
| 完整质粒大小: | 9144 bp |
| 载体类型: | E. coli - C. difficile shuttle vector |
| 载体抗性: | Chloramphenicol, 25 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
文献引用方法 |
|
| 材料和方法部分: | pJAK112 was a gift from Robert Fagan (Addgene plasmid # 167280 ; http://n2t.net/addgene:167280 ; RRID:Addgene_167280) |
| 参考文献部分: | An RNA-centric global view of Clostridioides difficile reveals broad activity of Hfq in a clinically important gram-positive bacterium. Fuchs , Manuela, Lamm-Schmidt, Vanessa, Sulzer , Johannes, Ponath, Falk, Jenniches , Laura, Kirk, Joseph A, Fagan, Robert P, Barquist, Lars, Vogel, Jörg, Faber, Franziska 5. Proc Natl Acad Sci USA, 2021, Jun 22;118(25) . 10.1073/pnas.2103579118 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pJAK112 9144 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION CodA-based mutagenesis vector for C. difficile.
ACCESSION .
VERSION .
KEYWORDS pJAK112
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9144)
TITLE An RNA-centric global view of Clostridioides difficile reveals broad
activity of Hfq in a clinically important Gram-positive bacterium
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9144)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9144
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 108..124
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 108..124
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
protein_bind 299..323
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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)."
RBS 342..350
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 366..1646
/codon_start=1
/gene="codA"
/product="E. coli cytosine deaminase"
/label=codA
/note="confers sensitivity to 5-fluorocytosine"
/translation="SNNALQTIINARLPGEEGLWQIHLQDGKISAIDAQSGVMPITENS
LDAEQGLVIPPFVEPHIHLDTTQTAGQPNWNQSGTLFEGIERWAERKALLTHDDVKQRA
WQTLKWQIANGIQHVRTHVDVSDATLTALKAMLEVKQEVAPWIDLQIVAFPQEGILSYP
NGEALLEEALRLGADVVGAIPHFEFTREYGVESLHKTFALAQKYDRLIDVHCDEIDDEQ
SRFVETVAALAHHEGMGARVTASHTTAMHSYNGAYTSRLFRLLKMSGINFVANPLVNIH
LQGRFDTYPKRRGITRVKEMLESGINVCFGHDDVFDPWYPLGTANMLQVLHMGLHVCQL
MGYGQINDGLNLITHHSARTLNLQDYGIAAGNSANLIILPAENGFDALRRQVPVRYSVR
GGKVIASTQPAQTTVYLEQPEAIDYKR"
primer_bind complement(1746..1763)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1746..1762)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1755..1777)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 7633..8221
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8122..8141
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8375..8392
/label=L4440
/note="L4440 vector, forward primer"
oriT 8542..8651
/note="incP origin of transfer"
CDS 8684..9055
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
ORIGIN
1 tggtgaagta catcaccgac gagcaaggca agaccgatcg ggccccctgc aggataaaaa
61 aattgtagat aaattttata aaatagtttt atctacaatt tttttatcag gaaacagcta
121 tgaccgcggc cgctgtatcc atatgaccat gattacgaat tccccggatc gagatagtat
181 atgatgcata ttctttaaat atagataaag ttatagaagc aatagaagat ttaggattta
241 ctgtaatata aattacactt ttaaaaagtt taaaaacatg atacaataag ttatggttgg
301 aattgttatc cgctcacaat tccaacttat gattaaaatt ttaaggaggt gtatttcata
361 atgtgtcgaa taacgcttta caaacaatta ttaacgcccg gttaccaggc gaagaggggc
421 tgtggcagat tcatctgcag gacggaaaaa tcagcgccat tgatgcgcaa tccggcgtga
481 tgcccataac tgaaaacagc ctggatgccg aacaaggttt agttataccg ccgtttgtgg
541 agccacatat tcacctggac accacgcaaa ccgccggaca accgaactgg aatcagtccg
601 gcacgctgtt tgaaggcatt gaacgctggg ccgagcgcaa agcgttatta acccatgacg
661 atgtgaaaca acgcgcatgg caaacgctga aatggcagat tgccaacggc attcagcatg
721 tgcgtaccca tgtcgatgtt tcggatgcaa cgctaactgc gctgaaagca atgctggaag
781 tgaagcagga agtcgcgccg tggattgatc tgcaaatcgt cgccttccct caggaaggga
841 ttttgtcgta tcccaacggt gaagcgttgc tggaagaggc gttacgctta ggggcagatg
901 tagtgggggc gattccgcat tttgaattta cccgtgaata cggcgtggag tcgctgcata
961 aaaccttcgc cctggcgcaa aaatacgacc gtctcatcga cgttcactgt gatgagatcg
1021 atgacgagca gtcgcgcttt gtcgaaaccg ttgctgccct ggcgcaccat gaaggcatgg
1081 gcgcacgagt caccgccagc cacaccacgg caatgcactc ctataacggg gcgtatacct
1141 cacgcctgtt ccgcttgctg aaaatgtccg gtattaactt tgtcgccaac ccgctggtca
1201 atattcatct gcaaggacgt ttcgatacgt atccaaaacg tcgcggcatc acgcgcgtta
1261 aagagatgct ggagtccggc attaacgtct gctttggtca cgatgatgtc ttcgatccgt
1321 ggtatccgct gggaacggcg aatatgctgc aagtgctgca tatggggctg catgtttgcc
1381 agttgatggg ctacgggcag attaacgatg gcctgaattt aatcacccac cacagcgcaa
1441 ggacgttgaa tttgcaggat tacggcattg ccgccggaaa cagcgccaac ctgattatcc
1501 tgccggctga aaatgggttt gatgcgctgc gccgtcaggt tccggtacgt tattcggtac
1561 gtggcggcaa ggtgattgcc agcacacaac cggcacaaac caccgtatat ctggagcagc
1621 cagaagccat cgattacaaa cgttgaacga ctgggttaca gcgagcttag tttaaagggc
1681 gaattcctag agtcgacgtc acgcgtccat ggagatctcg aggcctgcag acatgcaagc
1741 ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
1801 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
1861 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctagca taaaaataag
1921 aagcctgcat ttgcaggctt cttattttta tggcgcgccg ttctgaatcc ttagctaatg
1981 gttcaacagg taactatgac gaagatagca ccctggataa gtctgtaatg gattctaagg
2041 catttaatga agacgtgtat ataaaatgtg ctaatgaaaa agaaaatgcg ttaaaagagc
2101 ctaaaatgag ttcaaatggt tttgaaattg attggtagtt taatttaata tattttttct
2161 attggctatc tcgataccta tagaatcttc tgttcacttt tgtttttgaa atataaaaag
2221 gggcttttta gccccttttt tttaaaactc cggaggagtt tcttcattct tgatactata
2281 cgtaactatt tcgatttgac ttcattgtca attaagctag taaaatcaat ggttaaaaaa
2341 caaaaaactt gcatttttct acctagtaat ttataatttt aagtgtcgag tttaaaagta
2401 taatttacca ggaaaggagc aagtttttta ataaggaaaa atttttcctt ttaaaattct
2461 atttcgttat atgactaatt ataatcaaaa aaatgaaaat aaacaagagg taaaaactgc
2521 tttagagaaa tgtactgata aaaaaagaaa aaatcctaga tttacgtcat acatagcacc
2581 tttaactact aagaaaaata ttgaaaggac ttccacttgt ggagattatt tgtttatgtt
2641 gagtgatgca gacttagaac attttaaatt acataaaggt aatttttgcg gtaatagatt
2701 ttgtccaatg tgtagttggc gacttgcttg taaggatagt ttagaaatat ctattcttat
2761 ggagcattta agaaaagaag aaaataaaga gtttatattt ttaactctta caactccaaa
2821 tgtaaaaagt tatgatctta attattctat taaacaatat aataaatctt ttaaaaaatt
2881 aatggagcgt aaggaagtta aggatataac taaaggttat ataagaaaat tagaagtaac
2941 ttaccaaaag gaaaaataca taacaaagga tttatggaaa ataaaaaaag attattatca
3001 aaaaaaagga cttgaaattg gtgatttaga acctaatttt gatacttata atcctcattt
3061 tcatgtagtt attgcagtta ataaaagtta ttttacagat aaaaattatt atataaatcg
3121 agaaagatgg ttggaattat ggaagtttgc tactaaggat gattctataa ctcaagttga
3181 tgttagaaaa gcaaaaatta atgattataa agaggtttac gaacttgcga aatattcagc
3241 taaagacact gattatttaa tatcgaggcc agtatttgaa attttttata aagcattaaa
3301 aggcaagcag gtattagttt ttagtggatt ttttaaagat gcacacaaat tgtacaagca
3361 aggaaaactt gatgtttata aaaagaaaga tgaaattaaa tatgtctata tagtttatta
3421 taattggtgc aaaaaacaat atgaaaaaac tagaataagg gaacttacgg aagatgaaaa
3481 agaagaatta aatcaagatt taatagatgg aatagaaata gattaaagtg taactatact
3541 ttatatatat atgattaaaa aaataaaaaa caacagccta ttaggttgtt gttttttatt
3601 ttctttatta atttttttaa tttttagttt ttagttcttt tttaaaataa gtttcagcct
3661 ctttttcaat attttttaaa gaaggagtat ttgcatgaat tgcctttttt ctaacagact
3721 taggaaatat tttaacagta tcttcttgcg ccggtgattt tggaacttca taacttacta
3781 atttataatt attattttct tttttaattg taacagttgc aaaagaagct gaacctgttc
3841 cttcaactag tttatcatct tcaatataat attcttgacc tatatagtat aaatatattt
3901 ttattatatt tttacttttt tctgaatcta ttattttata atcataaaaa gttttaccac
3961 caaaagaagg ttgtactcct tctggtccaa catatttttt tactatatta tctaaataat
4021 ttttgggaac tggtgttgta atttgattaa tcgaacaacc agttatactt aaaggaatta
4081 taactataaa aatatatagg attatctttt taaatttcat tattggcctc ctttttatta
4141 aatttatgtt accataaaaa ggacataacg ggaatatgta gaatattttt aatgtagaca
4201 aaattttaca taaatataaa gaaaggaagt gtttgtttaa attttatagc aaactatcaa
4261 aaattagggg gataaaaatt tatgaaaaaa aggttttcga tgttattttt atgtttaact
4321 ttaatagttt gtggtttatt tacaaattcg gccggccagt gggcaagttg aaaaattcac
4381 aaaaatgtgg tataatatct ttgttcatta gagcgataaa cttgaatttg agagggaact
4441 tagatggtat ttgaaaaaat tgataaaaat agttggaaca gaaaagagta ttttgaccac
4501 tactttgcaa gtgtaccttg tacctacagc atgaccgtta aagtggatat cacacaaata
4561 aaggaaaagg gaatgaaact atatcctgca atgctttatt atattgcaat gattgtaaac
4621 cgccattcag agtttaggac ggcaatcaat caagatggtg aattggggat atatgatgag
4681 atgataccaa gctatacaat atttcacaat gatactgaaa cattttccag cctttggact
4741 gagtgtaagt ctgactttaa atcattttta gcagattatg aaagtgatac gcaacggtat
4801 ggaaacaatc atagaatgga aggaaagcca aatgctccgg aaaacatttt taatgtatct
4861 atgataccgt ggtcaacctt cgatggcttt aatctgaatt tgcagaaagg atatgattat
4921 ttgattccta tttttactat ggggaaatat tataaagaag ataacaaaat tatacttcct
4981 ttggcaattc aagttcatca cgcagtatgt gacggatttc acatttgccg ttttgtaaac
5041 gaattgcagg aattgataaa tagttaactt caggtttgtc tgtaactaaa aacaagtatt
5101 taagcaaaaa catcgtagaa atacggtgtt ttttgttacc ctagagctct atacaggtga
5161 agcagatgct atgagtgcgt catcagtagc atcaagagat ggatcaccta taattttaac
5221 taatggtaaa aatgttccat ttgagaaaaa agaaggtgtg caatgttatg ctcttggttc
5281 agaggagata ataagtaatg atttagttaa aaaaactaat gcagttagat tagctggaga
5341 agatagattt gagactaata aaaaagttat aaaacatttt tatagttcgg caaaagaatt
5401 ttatctttct aaaggttatc aattagtaga tgcagttgct ggttcttcta tagcaaaaaa
5461 tgcacctatt gtattagttg acggaaatag tgataagtct gtacttagaa gtgctgataa
5521 gattacagca ctaggaggaa tagatgaaaa gacactagaa caatgtttaa gtgcatcatc
5581 actggatgca agtgcaccaa caataactgt tggaaattta aatatttatc aaggtgataa
5641 gtttgatata agtaaactga atatcgtagc aaaggatagc aatggtaacg atttgacacc
5701 agagcttata ggaaatattg atactgataa ggtgggtaaa tacaaagtta ctataaaagc
5761 tactgatatt ggtggaaaga ctacatcaat aagtgtagaa gtaaatgtac ttgaatataa
5821 gactaatgat atgaatagta gtgagtttaa aagaatggtt tctagtgaaa tgtatagttt
5881 ggtaaattca tatagaaaag aaaaaggtaa agagccatta caagtttcta aaaacttaga
5941 aggactttct aattcctggt ctaagtacat ggcagataaa ggtcagtttt cacatgtaat
6001 aaatggtaag aaatctgttg aagtatttac tggatatggc ttgagaagtg aagaaaatat
6061 agcttttgta ccacttgtta caaaaagtac ttatacaaca aaggatgcta gagaagtagc
6121 aaatgtaata tttacagttt ggaaaaagtc agataaatat aatgaaaata tgttaaatag
6181 tgattttgta tatactgggt ttggattata tatattgcct aatggagaag tgtatgctac
6241 acaagaattt ttaaataagt aatttaaata agtaaaatca aattttttaa gttattaaat
6301 attggagttt aagggtataa aaactcatat gcttgggtat aaatataata atataaaaag
6361 gcttcttata tgagaagtct ttttatatta tgttttcgaa tacaaaaaaa gagactaaat
6421 ttagtctctt ttttatgtga gaaatactta aatatataaa gatgtaattt atttaagtat
6481 gattattata ctatacaata aaataattgt ctactattct ataattttaa ggtataataa
6541 taattgtatg ataaaagagg aggtcaaaaa atatgtcagt tttagatgca atcttagata
6601 aagcggatga acaagaaatt aaaaaattaa atttaatagt agataaaata gaatctttag
6661 aaaaagatat ggaaatcaaa gagaatgaac aattaaaaga aatgacaaat acttttagac
6721 ttagattaga caaaggtgaa agtttggatg atatacttcc agaagcattt gctgttgtta
6781 gagaggtctc aaaaagagtt ttaggaatgc gtcaatataa agttcagtta attggaggta
6841 tagtaattca tcaaggaaag atagctgaga tgaagactgg tgaggggaaa actttagttg
6901 aggtagctcc agtatatcta aatgctttaa caggaaaagg tgtacatgta attacagtaa
6961 atgattattt ggcagaacgt gataaagaac ttatgcgtcc agtatatgaa tttttaggat
7021 tgactgtagg tgtaatttta tctaaccaag acccaagtat aagaaaagaa caatataaat
7081 gtgatataac ttatggaaca aatagtgaat ttggatttga ctatttaaga gataatatgg
7141 tgccagattt gtctcataaa gtacaaaggg aactaaactt tgccatagta gatgagatag
7201 actcaatatt aatagatgaa gctagaactc cacttattat tgcaggagat ggtgatgaag
7261 atttaaaact ttatgagtta gcaaataact ttgtaaaaac tgttaaagaa gaagactttg
7321 agatggatag aaaagataaa actatagcat taacagcaag tggtataagc aaagctgagt
7381 cattttttgg tataacaaac cttactgata taaagaatat agaattatat catcatataa
7441 atcaagcttt aagaggtcat aagcttatgg aaaaagatgt tgactatgtt atctcaaatg
7501 gagaagtaat gatagttgac gaatttacag gaagagggat cccctttttg ataatctcat
7561 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
7621 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
7681 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
7741 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
7801 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
7861 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
7921 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
7981 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
8041 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
8101 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
8161 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
8221 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
8281 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
8341 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
8401 agagcgccca atacgcaggg ccccctgctt cggggtcatt atagcgattt tttcggtata
8461 tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg
8521 gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg
8581 ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg
8641 aggctggccg gctaccgccg gcgtaacaga tgagggcaag cggatggctg atgaaaccaa
8701 gccaaccagg aagggcagcc cacctatcaa ggtgtactgc cttccagacg aacgaagagc
8761 gattgaggaa aaggcggcgg cggccggcat gagcctgtcg gcctacctgc tggccgtcgg
8821 ccagggctac aaaatcacgg gcgtcgtgga ctatgagcac gtccgcgagc tggcccgcat
8881 caatggcgac ctgggccgcc tgggcggcct gctgaaactc tggctcaccg acgacccgcg
8941 cacggcgcgg ttcggtgatg ccacgatcct cgccctgctg gcgaagatcg aagagaagca
9001 ggacgagctt ggcaaggtca tgatgggcgt ggtccgcccg agggcagagc catgactttt
9061 ttagccgcta aaacggccgg ggggtgcgcg tgattgccaa gcacgtcccc atgcgctcca
9121 tcaagaagag cgacttcgcg gagc
//
