| 质粒名称: | pJQ200mp18 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 78500 |
| 存储实验室: | Michael Hynes |
|
载体骨架基本信息 |
|
| 载体名称: | pJQ200 |
| 载体出品公司: | Jurgen Quandt |
| 载体抗性: | 庆大霉素 |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | -- |
| 载体修饰: | 来自pUC18的polylinker和lacZ alpha作为部分PvuI摘要添加到pJQ200中。 |
| 载体类型: | 用于基因替换的自杀载体 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pJQ200mp18 was a gift from Michael Hynes (Addgene plasmid # 78500 ; http://n2t.net/addgene:78500 ; RRID:Addgene_78500) |
| 参考文献部分: | Versatile suicide vectors which allow direct selection for gene replacement in gram-negative bacteria. Quandt J, Hynes MF. Gene. 1993 May 15;127(1):15-21. 0378-1119(93)90611-6 [pii] PubMed 8486283 |
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 pJQ200mp18 5871 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Suicide vector for gene replacement.
ACCESSION .
VERSION .
KEYWORDS pJQ200mp18
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5871)
AUTHORS Quandt J, Hynes MF
TITLE Versatile suicide vectors which allow direct selection for gene
replacement in gram-negative bacteria.
JOURNAL Gene. 1993 May 15;127(1):15-21.
PUBMED 8486283
REFERENCE 2 (bases 1 to 5871)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5871
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(491..519)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
protein_bind 1347..1368
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1383..1413
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1421..1437
/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)."
primer_bind 1426..1448
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 1445..1461
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 1445..1461
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
misc_feature 1470..1526
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(1530..1547)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1530..1546)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1539..1561)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(2133..2150)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2267..2812)
/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(2347..2366)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
oriT 3195..3304
/note="incP origin of transfer"
promoter 3680..4125
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 4126..5547
/codon_start=1
/gene="Bacillus subtilis sacB"
/product="secreted levansucrase that renders bacterial
growth sensitive to sucrose"
/label=SacB
/note="negative selection marker"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
CDS complement(5640..302)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
ORIGIN
1 tgctcaaact tgggcagaac gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag
61 gcagcaagcg cgatgaatgt cttactacgg agcaagttcc cgaggtaatc ggagtccggc
121 tgatgttggg agtaggtggc tacgtctccg aactcacgac cgaaaagatc aagagcagcc
181 cgcatggatt tgacttggtc agggccgagc ctacatgtgc gaatgatgcc catacttgag
241 ccacctaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct gctgcgtaac
301 atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt gctgcttgga
361 tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc
421 gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta
481 cttgcattac agtttacgaa ccgaacaggc ttatgtcaac tgggttcgtg ccttcatccg
541 tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg catttctgtc
601 ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat tggcggcctt
661 gctgttcttc tacggcaagg tgctgtgcac ggatctgccc tggcttcagg agatcggaag
721 acctcggccg tcgcggcgct tgccggtggt gctgaccccg gatgaagtgg ttcgcatcct
781 cggttttctg gaaggcgagc atcgtttgtt cgcccagctt ctgtatggaa cgggcatgcg
841 gatcagtgag ggtttgcaac tgcgggtcaa ggatctggat ttcgatcacg gcacgatcat
901 cgtgcgggag ggcaagggct ccaaggatcg ggccttgatg ttacccgaga gcttggcacc
961 cagcctgcgc gagcagaatt aattccctgg cgtaatagcg aagaggcccg caccgatcgc
1021 ccttcccaac agttgcgcag cctgaatggc gaatggcgat aagctagctt cacgctgccg
1081 caagcactca gggcgcaagg gctgctaaag gaagcggaac acgtagaaag ccagtccgca
1141 gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc
1201 aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc
1261 ggttttatgg acagcaagcg aaccggaatt gccagctggc acgacaggtt tcccgactgg
1321 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
1381 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
1441 cacacaggaa acagctatga catgattacg aattcgagct cggtacccgg ggatcctcta
1501 gagtcgacct gcaggcatgc aagcttggca ctggccgtcg ttttacaacg tcgtgactgg
1561 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
1621 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
1681 gaatggcgat aagctagctt cacgctgccg caagcactca gggcgcaagg gctgctaaag
1741 gaagcggaac acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag
1801 ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag
1861 tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt
1921 gccaggggaa ttaattctgt ccctcctgtt cagctactga cggggtggtg cgtaacggca
1981 aaagcaccgc cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga
2041 tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag
2101 cagaatatgt gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc
2161 gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc
2221 aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc
2281 gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag
2341 gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg
2401 cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg
2461 acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt
2521 cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat
2581 gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc
2641 atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca
2701 gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc
2761 ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca
2821 tcttattaat cagataaaat atttctagct agagtcgatc ttcgccagca gggcgaggat
2881 cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt tcagcaggcc
2941 gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg cggacgtgct catagtccac
3001 gacgcccgtg attttgtagc cctggccgac ggccagcagg taggccgaca ggctcatgcc
3061 ggccgccgcc gccttttcct caatcgctct tcgttcgtct ggaaggcagt acaccttgat
3121 aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt
3181 tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg
3241 cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct
3301 gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc
3361 tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca
3421 gggttatgca gcggaaaagc gctgcttccc tgctgttttg tggaatatct accgactgga
3481 aacaggcaaa tgcaggaaat tactgaactg aggggacagg cgagagacga tgccaaagag
3541 ctacaccgac gagctggccg agtgggttga atcccgcgcg gccaagaagc gccggcgtga
3601 tgaggctgcg gttgcgttcc tggcggtgag ggcggatgtc gatcgactct agctagagga
3661 tcgatccttt ttaacccatc acatatacct gccgttcact attatttagt gaaatgagat
3721 attatgatat tttctgaatt gtgattaaaa aggcaacttt atgcccatgc aacagaaact
3781 ataaaaaata cagagaatga aaagaaacag atagattttt tagttcttta ggcccgtagt
3841 ctgcaaatcc ttttatgatt ttctatcaaa caaaagagga aaatagacca gttgcaatcc
3901 aaacgagagt ctaatagaat gaggtcgaaa agtaaatcgc gcgggtttgt tactgataaa
3961 gcaggcaaga cctaaaatgt gtaaagggca aagtgtatac tttggcgtca ccccttacat
4021 attttaggtc tttttttatt gtgcgtaact aacttgccat cttcaaacag gagggctgga
4081 agaagcagac cgctaacaca gtacataaaa aaggagacat gaacgatgaa catcaaaaag
4141 tttgcaaaac aagcaacagt attaaccttt actaccgcac tgctggcagg aggcgcaact
4201 caagcgtttg cgaaagaaac gaaccaaaag ccatataagg aaacatacgg catttcccat
4261 attacacgcc atgatatgct gcaaatccct gaacagcaaa aaaatgaaaa atatcaagtt
4321 cctgaattcg attcgtccac aattaaaaat atctcttctg caaaaggcct ggacgtttgg
4381 gacagctggc cattacaaaa cgctgacggc actgtcgcaa actatcacgg ctaccacatc
4441 gtctttgcat tagccggaga tcctaaaaat gcggatgaca catcgattta catgttctat
4501 caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg ctggccgcgt ctttaaagac
4561 agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc aaacacaaga atggtcaggt
4621 tcagccacat ttacatctga cggaaaaatc cgtttattct acactgattt ctccggtaaa
4681 cattacggca aacaaacact gacaactgca caagttaacg tatcagcatc agacagctct
4741 ttgaacatca acggtgtaga ggattataaa tcaatctttg acggtgacgg aaaaacgtat
4801 caaaatgtac agcagttcat cgatgaaggc aactacagct caggcgacaa ccatacgctg
4861 agagatcctc actacgtaga agataaaggc cacaaatact tagtatttga agcaaacact
4921 ggaactgaag atggctacca aggcgaagaa tctttattta acaaagcata ctatggcaaa
4981 agcacatcat tcttccgtca agaaagtcaa aaacttctgc aaagcgataa aaaacgcacg
5041 gctgagttag caaacggcgc tctcggtatg attgagctaa acgatgatta cacactgaaa
5101 aaagtgatga aaccgctgat tgcatctaac acagtaacag atgaaattga acgcgcgaac
5161 gtctttaaaa tgaacggcaa atggtacctg ttcactgact cccgcggatc aaaaatgacg
5221 attgacggca ttacgtctaa cgatatttac atgcttggtt atgtttctaa ttctttaact
5281 ggcccataca agccgctgaa caaaactggc cttgtgttaa aaatggatct tgatcctaac
5341 gatgtaacct ttacttactc acacttcgct gtacctcaag cgaaaggaaa caatgtcgtg
5401 attacaagct atatgacaaa cagaggattc tacgcagaca aacaatcaac gtttgcgcca
5461 agcttcctgc tgaacatcaa aggcaagaaa acatctgttg tcaaagacag catccttgaa
5521 caaggacaat taacagttaa caaataaaaa cgcaaaagaa aatgccgatg gccgcggcgt
5581 tgtgacaatt taccgaacaa ctccgcggcc gggaagccga tctcggcttg aacgaattgt
5641 taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat gcccaacttt
5701 gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat cacataagca
5761 ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat gccctgcctc
5821 cggtgctcgc cggagactgc gagatcatag atatagatct cactacgcgg c
//
