| 质粒名称: | pGRB |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 71539 |
| 存储实验室: | Tao Chen |
|
载体骨架基本信息 |
|
| 载体名称: | bonepuc18 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 2705bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 5’测序引物 | ACTAGTATTATACCTAGGACTGAGC |
| 3’测序引物 | GTTTTAGAGCTAGAAATAGCAAGTTAA |
|
文献引用方法 |
|
| 材料和方法部分: | pGRB was a gift from Tao Chen (Addgene plasmid # 71539 ; http://n2t.net/addgene:71539 ; RRID:Addgene_71539) |
| 参考文献部分: | Metabolic engineering of Escherichia coli using CRISPR-Cas9 meditated genome editing. Li Y, Lin Z, Huang C, Zhang Y, Wang Z, Tang YJ, Chen T, Zhao X. Metab Eng. 2015 Sep;31:13-21. doi: 10.1016/j.ymben.2015.06.006. Epub 2015 Jun 30. 10.1016/j.ymben.2015.06.006 PubMed 26141150 |
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 pGRB 2705 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION for expression of gRNA in E. coli.
ACCESSION .
VERSION .
KEYWORDS pGRB
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2705)
AUTHORS Li Y, Lin Z, Huang C, Zhang Y, Wang Z, Tang YJ, Chen T, Zhao X
TITLE Metabolic engineering of Escherichia coli using CRISPR-Cas9
meditated genome editing.
JOURNAL Metab Eng. 2015 Sep;31:13-21. doi: 10.1016/j.ymben.2015.06.006. Epub
2015 Jun 30.
PUBMED 26141150
REFERENCE 2 (bases 1 to 2705)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..2705
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..35
/label=J23119(SpeI) promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119) modified to end with an SpeI site"
primer_bind complement(59..81)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(176..493)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNRLA
AHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDA
A"
primer_bind 181..200
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 394..416
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 408..425
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 409..425
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 429..485
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(494..510)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(494..510)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(507..529)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 518..534
/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(542..572)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 587..608
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(724..741)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(895..1483)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(975..994)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1654..2514)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 2277..2296
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2515..2619)
/gene="bla"
/label=AmpR promoter
primer_bind 2687..2705
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 ttgacagcta gctcagtcct aggtataata ctagtttttt cggtgatgac ggtgaaaacc
61 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
121 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
181 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
241 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
301 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
361 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
421 ccagtgccaa gcttgcatgc ctgcaggtcg actctagagg atccccgggt accgagctcg
481 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
541 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
601 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcagttttag
661 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccc
721 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
781 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
841 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
901 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
961 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
1021 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
1081 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
1141 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
1201 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
1261 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
1321 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
1381 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
1441 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
1501 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
1561 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
1621 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
1681 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
1741 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgtgaccc
1801 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
1861 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
1921 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
1981 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
2041 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
2101 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
2161 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
2221 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
2281 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
2341 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
2401 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
2461 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
2521 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
2581 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
2641 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
2701 gaggc
//
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