| 质粒名称: | pJQ200SK |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 78497 |
| 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: | -- |
| 载体抗性: | 庆大霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 4922 bp |
| 载体修饰: | 添加多克隆位点和部分lacZα基因作为PvuII片段克隆到pJQ200的SmaI位点 |
| 载体类型: | 自杀质粒 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 未知 |
| 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 | T7 |
| 3' 测序引物 | T3 |
|
文献引用方法 |
|
| 材料和方法部分: | pJQ200SK was a gift from Michael Hynes (Addgene plasmid # 78497 ; http://n2t.net/addgene:78497 ; RRID:Addgene_78497) |
| 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 pJQ200SK 5217 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Suicide vector for gene replacement.
ACCESSION .
VERSION .
KEYWORDS pJQ200SK
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5217)
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 5217)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5217
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(37..53)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(37..53)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(90..110)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(90..108)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(129..145)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(129..145)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(142..164)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 153..169
/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)."
promoter complement(177..207)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 222..243
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(502..519)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(636..1180)
/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(716..735)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
oriT 1554..1663
/note="incP origin of transfer"
promoter 2039..2484
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 2485..3909
/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
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYRGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGISSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTS
VVKDSILEQGQLTVNK"
CDS complement(4002..4535)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGSEFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(4724..4752)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
ORIGIN
1 tcgataagct tgatatcgaa ttcctgcagc ccgggggatc cactagttct agagcggccg
61 ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
121 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
181 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
241 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccaggggaat
301 taattctgtc cctcctgttc agctactgac ggggtggtgc gtaacggcaa aagcaccgcc
361 ggacatcagc gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca
421 gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg
481 atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
541 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
601 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
661 aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
721 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
781 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
841 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agttcgaccg
901 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
961 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
1021 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
1081 tcaaagagtt ggtagctcag agaaccttcg aaaaccgccc tgcaaggcgg ttttttcgtt
1141 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
1201 gataaaatat ttctagctag agtcgatctt cgccagcagg gcaggatcgt ggcatcaccg
1261 aaccgcgccg tgccgctcgt cggtgagcca gagtttcagc aggccgccca gggggaggtc
1321 gccattgatg cgggccagct cgcggacgtg ctcatagtcc acgacgcccg tgattttgta
1381 gccctggccg acgcgacagg taggccgaca gctcatgccg gccgccgccg ccttttcctc
1441 aatcgctctt cgttcgtctg gaaggcagta caccttgata ggtgggctgc ccttcctggt
1501 tggcttggtt tcatcagcca tccgcttgcc ctcatctgtt acgccggcgg tagccggcca
1561 gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa
1621 ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat
1681 acaccaagga aagtctacgc gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga
1741 tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag cggaaaagcg
1801 ctgcttccct gctgttttgt ggaatatcta ccgactggaa acaggcaaat gcaggaaatt
1861 actgaactga ggggacaggc gagagacgat gccaaagagc tacaccgacg agctggccga
1921 gtgggttgaa tcccgcgcgg ccaagaagcg ccggcgtgat gaggctgcgg ttgcgttcct
1981 ggcggtgagg gcggatgtcg atcgactcta gctagaggat cgatcctttt taacccatca
2041 catatacctg ccgttcacta ttatttagtg aaatgagata ttatgatatt ttctgaattg
2101 tgattaaaaa ggcaacttta tgcccatgca acagaaacta taaaaaatac agagaatgaa
2161 aagaaacaga tagatttttt agttctttag gcccgtagtc tgcaaatcct tttatgattt
2221 tctatcaaac aaaagaggaa aatagaccag ttgcaatcca aacgagagtc taatagaatg
2281 aggtcgaaaa gtaaatcgcg cgggtttgtt actgataaag caggcaagac ctaaaatgtg
2341 taaagggcaa agtgtatact ttggcgtcac cccttacata ttttaggtct ttttttattg
2401 tgcgtaacta acttgccatc ttcaaacagg agggctggaa gaagcagacc gctaacacag
2461 tacataaaaa aggagacatg aacgatgaac atcaaaaagt ttgcaaaaca agcaacagta
2521 ttaaccttta ctaccgcact gctggcagga ggcgcaactc aagcgtttgc gaaagaaacg
2581 aaccaaaagc catataagga aacatacggc atttcccata ttacacgcca tgatatgctg
2641 caaatccctg aacagcaaaa aaatgaaaaa tatcaagttc ctgaattcga ttcgtccaca
2701 attaaaaata tctcttctgc aaaaggcctg gacgtttggg acagctggcc attacaaaac
2761 gctgacggca ctgtcgcaaa ctatcgcggc taccacatcg tctttgcatt agccggagat
2821 cctaaaaatg cggatgacac atcgatttac atgttctatc aaaaagtcgg cgaaacttct
2881 attgacagct ggaaaaacgc tggccgcgtc tttaaagaca gcgacaaatt cgatgcaaat
2941 gattctatcc taaaagacca aacacaagaa tggtcaggtt cagccacatt tacatctgac
3001 ggaaaaatcc gtttattcta cactgatttc tccggtaaac attacggcaa acaaacactg
3061 acaactgcac aagttaacgt atcagcatca gacagctctt tgaacatcaa cggtgtagag
3121 gattataaat caatctttga cggtgacgga aaaacgtatc aaaatgtaca gcagttcatc
3181 gatgaaggca actacagctc aggcgacaac catacgctga gagatcctca ctacgtagaa
3241 gataaaggcc acaaatactt agtatttgaa gcaaacactg gaactgaaga tggctaccaa
3301 ggcgaagaat ctttatttaa caaagcatac tatggcaaaa gcacatcatt cttccgtcaa
3361 gaaagtcaaa aacttctgca aagcgataaa aaacgcacgg ctgagttagc aaacggcgct
3421 ctcggtatga ttgagctaaa cgatgattac acactgaaaa aagtgatgaa accgctgatt
3481 gcatctaaca cagtaacaga tgaaattgaa cgcgcgaacg tctttaaaat gaacggcaaa
3541 tggtacctgt tcactgactc ccgcggatca aaaatgacga ttgacggcat ttcgtctaac
3601 gatatttaca tgcttggtta tgtttctaat tctttaactg gcccatacaa gccgctgaac
3661 aaaactggcc ttgtgttaaa aatggatctt gatcctaacg atgtaacctt tacttactca
3721 cacttcgctg tacctcaagc gaaaggaaac aatgtcgtgg tgattacaag ctatatgaca
3781 aacagaggat tctacgcaga caaacaatca acgtttgcgc caagcttcct gctgaacatc
3841 aaaggcaaga aaacatctgt tgtcaaagac agcatccttg aacaaggaca attaacagtt
3901 aacaaataaa aacgcaaaag aaaatgccga tggccgcggc gttgtgacaa tttaccgaac
3961 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
4021 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
4081 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
4141 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
4201 gcgagatcat agatatagat ctcactacgc ggctgctcaa acctgggcag aacgtaagcc
4261 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
4321 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
4381 ccgaactcac taccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
4441 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
4501 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatgaaaca
4561 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
4621 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
4681 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
4741 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
4801 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
4861 ggtctccacg catcgtcagg cattggcggc caagctgttc ttctacggca aggtgctgtg
4921 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt
4981 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
5041 gttcgcccag cttctgtatg gaacgggcat gcggatcagt gggtttgcaa ctgcgggtca
5101 aggtctggat ttcgatcacg gcacgatcat gctgcgggag ggcaagggct ccaaggatcg
5161 ggccttgatg ttacccgaga gcttggcacc cacgctgcgc gagcagaatt aattccc
//
