| 质粒名称: | pGPI-SceI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 32060 |
| 存储实验室: | Miguel Valvano |
|
载体骨架基本信息 |
|
| 载体名称: | pGPI-oriR6K |
| 载体出品公司: | Valvano |
| 载体类型: | 诱变质粒 |
| 筛选标记: | 甲氧苄啶 |
| 载体抗性: | 甲氧苄啶 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Pir1 ; any strain containing lambda pir, Pir1 |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | TAACGGTTGTGGACAACAAGCCAGGG |
| 3’测序引物 | GCCCTACACAAATTGGGAGATATATC |
|
文献引用方法 |
|
| 材料和方法部分: | pGPI-SceI was a gift from Miguel Valvano (Addgene plasmid # 32060 ; http://n2t.net/addgene:32060 ; RRID:Addgene_32060) |
| 参考文献部分: | A system for the construction of targeted unmarked gene deletions in the genus Burkholderia. Flannagan RS, Linn T, Valvano MA. Environ Microbiol. 2008 Jun;10(6):1652-60. Epub 2008 Mar 13. 10.1111/j.1462-2920.2008.01576.x PubMed 18341581 |
买家导航
pGPISceI载体基本信息
pGPISceI载体图谱质粒图谱和多克隆位点信息
pGPISceI载体简介
pGPISceI载体序列
LOCUS pGPI-SceI 4559 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Construction of unmarked gene deletions. Suicide cloning vector
designed to introduce a targeted I-SceI restriction site in the
genome of Burkholderia.
ACCESSION .
VERSION .
KEYWORDS pGPI-SceI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4559)
AUTHORS Flannagan RS, Linn T, Valvano MA
TITLE A system for the construction of targeted unmarked gene deletions in
the genus Burkholderia.
JOURNAL Environ Microbiol. 2008 Jun;10(6):1652-60. Epub 2008 Mar 13.
PUBMED 18341581
REFERENCE 2 (bases 1 to 4559)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4559
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1..237
/codon_start=1
/gene="dhfrIIb from plasmid R388"
/product="E. coli plasmid-associated dihydrofolate
reductase"
/label=TpR
/note="confers resistance to trimethoprim"
/translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
primer_bind complement(873..892)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1033..1055
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 1194..1213
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
oriT 1894..2003
/note="incP origin of transfer"
CDS 2036..2407
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
rep_origin 3025..3413
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
protein_bind 3448..3464
/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 4206..4234
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
ORIGIN
1 atgggtcaaa gtagcgatga agccaacgct cccgttgcag ggcagtttgc gcttcccctg
61 agtgccacct ttggcttagg ggatcgcgta cgcaagaaat ctggtgccgc ttggcagggt
121 caagtcgtcg gttggtattg cacaaaactc actcctgaag gctatgcggt cgagtccgaa
181 tcccacccag gctcagtgca aatttatcct gtggctgcac ttgaacgtgt ggcctaagca
241 ttaccctgtt atccctaact gcactcacca gtcacagaaa agcatcttac ggatggcatg
301 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
361 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
421 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
481 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
541 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
601 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
661 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
721 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
781 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
841 atactttaga ttgatttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
901 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
961 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
1021 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
1081 cgaggcagca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
1141 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
1201 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
1261 ccggcgtaga ggatcctttt tgtccggtgt tgggttgaag gtgaagccgg tcggggccgc
1321 agcgggggcc ggcttttcag ccttgccccc ctgcttcggc cgccgtggct ccggcgtctt
1381 gggtgccggc gcgggttccg cagccttggc ctgcggtgcg ggcacatcgg cgggcttggc
1441 cttgatgtgc cgcctggcgt gcgagcggaa cgtctcgtag gagaacttga ccttccccgt
1501 ttcccgcatg tgctcccaaa tggtgacgag cgcatagccg gacgctaacg ccgcctcgac
1561 atccgccctc accgccagga acgcaaccgc agcctcatca cgccggcgct tcttggccgc
1621 gcgggattca acccactcgg ccagctcgtc ggtgtagctc tttggcatcg tctctcgcct
1681 gtcccctcag ttcagtaatt tcctgcattt gcctgtttcc agtcggtaga tattccacaa
1741 aacagcaggg aagcagcgct tttccgctgc ataaccctgc ttcggggtca ttatagcgat
1801 tttttcggta tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt
1861 agactttcct tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc
1921 cgcgagcggg tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa
1981 tcctgctctg cgaggctggc cggctaccgc cggcgtaaca gatgagggca agcggatggc
2041 tgatgaaacc aagccaacca ggaagggcag cccacctatc aaggtgtact gccttccaga
2101 cgaacgaaga gcgattgagg aaaaggcggc ggcggccggc atgagcctgt cggcctacct
2161 gctggccgtc ggccagggct acaaaatcac gggcgtcgtg gactatgagc acgtccgcga
2221 gctggcccgc atcaatggcg acctgggccg cctgggcggc ctgctgaaac tctggctcac
2281 cgacgacccg cgcacggcgc ggttcggtga tgccacgatc ctcgccctgc tggcgaagat
2341 cgaagagaag caggacgagc ttggcaaggt catgatgggc gtggtccgcc cgagggcaga
2401 gccatgactt ttttagccgc taaaacggcc ggggggtgcg cgtgattgcc aagcacgtcc
2461 ccatgcgctc catcaagaag agcgacttcg cggagctggt gaagtacatc accgacgagc
2521 aaggcaagac cgagcgcctg ggtcacgtgc gcgtcacgaa ctgcgaggca aacaccctgc
2581 ccgctgtcat ggccgaggtg atggcgaccc agcacggcaa cacccgttcc gaggccgaca
2641 agacctatca cctgctggtt agcttccgcg cgggagagaa gcccgacgcg gagacgttgc
2701 gcgcgattga ggaccgcatc tgcgctgggc ttggcttcgc cgagcatcag cgcgtcagtg
2761 ccgtgcatca cgacaccgac aacctgcaca tccatatcgc catcaacaag attcacccga
2821 cccgaaacac catccatgag ccgtatcggg cctaccgcgc cctcgctgac ctctgcgcga
2881 cgctcgaacg ggactacggg cttgagcgtg acaatcacga aacgcggcag cgcgtttccg
2941 agaaccgcgc gaacgacatg gagcggcacg cgggcgtgga aagcctggtc ggctggatcc
3001 ggccacgatg cgtccggcgt agaggatctg aagatcagca gttcaacctg ttgatagtac
3061 gtactaagct ctcatgtttc acgtactaag ctctcatgtt taacgtacta agctctcatg
3121 tttaacgaac taaaccctca tggctaacgt actaagctct catggctaac gtactaagct
3181 ctcatgtttc acgtactaag ctctcatgtt tgaacaataa aattaatata aatcagcaac
3241 ttaaatagcc tctaaggttt taagttttat aagaaaaaaa agaatatata aggcttttaa
3301 agcttttaag gtttaacggt tgtggacaac aagccaggga tgtaacgcac tgagaagccc
3361 ttagagcctc tcaaagcaat tttgagtgac acaggaacac ttaacggctg acatgggaat
3421 tccacatgtg gaattccaca tgtggaattg tgagcggata acaatttgtg gaattcccgg
3481 gagagctcga tatcgcatgc ggtacctcta gaagaagctt gggatccgtc ggggatccgg
3541 tgattgattg agcaagcttt atgcttgtaa accgttttgt gaaaaaattt ttaaaataaa
3601 aaaggggacc tctagggtcc ccaattaatt agtaatataa tctattaaag gtcattcaaa
3661 aggtcatcca ccggatcagc ttagtaaagc cctcgctaga ttttaatgcg gatgttgcga
3721 ttacttcgcc aactattgcg ataacaagaa aaagccagcc tttcatgata tatctcccaa
3781 tttgtgtagg gcttattatg cacgcttaaa aataataaaa gcagacttga cctgatagtt
3841 tggctgtgag caattatgtg cttagtgcat ctaacgcttg agttaagccg cgccgcgaag
3901 cggcgtcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt gatctcgcct
3961 ttcacgtagt ggacaaattc ttccaactga tctagcttta atgcggtagt ttatcacagt
4021 taaattgcta acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc
4081 tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc
4141 tcttgcggga ttctacgggg tctgacgctc agtggaacga aatcgatgag ctcgcacgaa
4201 cccagttgac ataagcctgt tcggttcgta aactgtaatg caagtagcgt atgcgctcac
4261 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
4321 tggcttgtta tgactgtttt tttgtacagt ctatgcctcg ggcatccaag cagcaagcgc
4381 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
4441 gtcgccctaa aacaaagtta ggcagccgtt gtgctggtgc tttctgatag ttgttgtggg
4501 gtaggcagtc agagctcgat ttgcttgtcg ccataataga ttcacaagaa ggattcgac
//
