| 质粒名称: | pTTCC |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 164264 |
| 存储实验室: | Gudmundur Hreggvidsson |
|
载体骨架基本信息 |
|
| 载体名称: | pLEI250.2 |
| 载体出品公司: | Lei Wang, Universitat Stuttgart |
| 空载体大小: | 5371 bp |
| 完整质粒大小: | 7778 bp |
| 载体修饰: | BbsI限制站点已删除 |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | Cas9 |
| 物种来源: | Synthetic; Synthesized, codon optimized Cas9 gene from Geobacillus sp. LC300 |
| 插入大小: | 3264 bp |
| 基因突变: | -- |
| GenBank编号 | AKU27017.1 |
| 启动子: | Pcel |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NdeI(未破坏) |
| 3’克隆位点: | HindIII(未破坏) |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pTTCC was a gift from Gudmundur Hreggvidsson (Addgene plasmid # 164264 ; http://n2t.net/addgene:164264 ; RRID:Addgene_164264) |
| 参考文献部分: | Efficient genome editing of an extreme thermophile, Thermus thermophilus, using a thermostable Cas9 variant. Adalsteinsson, Bjorn Thor, Kristjansdottir, Thordis, Merre, William, Helleux, Alexandra, Dusaucy, Julia, Tourigny, Mathilde, Fridjonsson, Olafur & Hreggvidsson, Gudmundur Oli. Scientific Reports volume 11, Article number: 9586 (2021) 10.1038/s41598-021-89029-2 |
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 pTTCC 7778 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses a thermostable Cas9 enzyme and gRNA in Thermus
thermophilus.
ACCESSION .
VERSION .
KEYWORDS pTTCC
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7778)
AUTHORS Adalsteinsson, Bjorn Thor, Kristjansdottir, Thordis, Merre, William,
Helleux, Alexandra, Dusaucy, Julia, Tourigny, Mathilde, Fridjonsson,
TITLE Efficient genome editing of an extreme thermophile, Thermus
thermophilus, using a thermostable Cas9 variant
JOURNAL Scientific Reports volume 11, Article number: 9586 (2021)
REFERENCE 2 (bases 1 to 7778)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7778
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(3485..3502)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 3655..3726
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
RBS 3912..3920
/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)"
promoter 6564..6593
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
RBS 6608..6616
/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 6622..6795
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGE"
primer_bind complement(6642..6659)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6642..6658)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6651..6673)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin complement(7147..7735)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7227..7246)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 acgaaaactc acgttaaggg attttggtca tgaattcgct agagtcgacc ccggggcgcg
61 aggggaggag ggctcgaggc ccccctcttg caagcgcttt caacagatgc tatactcccg
121 ggcaaagatg accaaggctc gccccaccat cgccgaaact agttcttgga ggagaaacgc
181 atatgcgcta caagatcggc ctggacatcg gcatcaccag cgtgggctgg gcggtgatca
241 acctcgatat tccgcggatc gaggacctgg gtgtccggat tttcgatcgc gccgaaaatc
301 cgcaaacggg agaaagcctg gccctgcccc gtcggctggc gcgcagcgcc cggcgtcgcc
361 tccgccgccg gaaacatcgc ctggaacgca ttcgccgtct gatcatccgg gaagggatcc
421 tgacgaagga ggagctggac aagctgtttg aggagaagca cgagatcgat gtctggcagc
481 tccgggtgga ggcgctggac cggaagctca acaacgacga gctggcgcgg gtcctgctgc
541 atctggccaa gcgccggggc ttcaaaagca accgcaagtc ggaacgctcg aacaaggaga
601 actcgacgat gctgaagcac atcgaggaga accgcgccat cctgtcctcg tatcgcaccg
661 tcggcgagat gatcgtgaaa gacccgaagt ttgccctgca caaacgtaac aagggcgaga
721 actacaccaa taccattgcc cgcgacgacc tggagcggga gatccggctc atcttctcga
781 agcagcgcga atttgggaac atgtcgtgca ccgaggagtt cgagaacgaa tacattgcca
841 tctgggcctc gcagcggccg gtggcgtcga aggacgacat tgagaagaag gtggggttct
901 gcacgttcga accgaaggag aagcgggcgc cgaaagccac gtacacgttc cagtcgttca
961 tcgcgtggga gcacatcaac aaactccggc tgatctcgcc gtcggggacc cgcggcctga
1021 cggatgagga acggcgcctc ctgtacgagc aggccttcca gaagaacaag atcacgtacc
1081 acgacatccg cacgctgctg cacctgccgg atgatacgta ttttaagggc attgtgtatg
1141 accgtggcga gtcgcggaaa cagaacgaga acatccggtt cctggagctc gacgcgtacc
1201 accaaatccg gaaggccgtg gacaaagtgt acggcaaagg caaatcgtcc tccttcctgc
1261 cgatcgactt cgataccttc ggatatgccc tgaccctgtt caaagacgac gcggacattc
1321 ggagctatct ccggaacgaa tacgagcaga acggtaaacg catgccgaac ctggccaaca
1381 aggtctacga caatgaactc atcgaagaac tgctcaacct gtccttcacg aagtttggcc
1441 acctgagcct gaaggccctg cgcagcatcc tcccctacat ggagcagggc gaagtgtaca
1501 gctcggcctg tgagcgcgcg ggatacacct tcacgggccc caagaagaag caaaagacga
1561 tgctgctgcc gaacattccg cccatcgcca accccgtggt catgcgggcc ctgacgcagg
1621 cccggaaggt ggtgaacgcg atcatcaaga agtacggaag ccccgtgtcg atccacatcg
1681 aactggcccg ggacctgtcg cagacgttcg acgaacgccg caaaaccaag aaggaacagg
1741 atgagaaccg caagaagaac gaaacggcca tccgccagct gatggagtac ggactgacgc
1801 tgaatccgac gggtcacgac atcgtgaaat tcaaactgtg gtcggagcag aacggccgct
1861 gcgcctattc gctgcagccc atcgaaatcg agcggctcct ggaaccgggc tacacggagg
1921 tggaccacgt cattccgtac tcgcgctccc tggatgactc ctacaccaac aaggtgctgg
1981 tcctcacgaa ggagaatcgc gagaagggta accgcatccc ggccgagtat ctgggcgtgg
2041 gcacggagcg ctggcagcag ttcgagacgt ttgtcctgac gaacaaacag ttcagcaaga
2101 agaaacgcga ccggctgctg cgcctgcatt acgacgaaaa cgaggaaacg gagttcaaga
2161 accgcaacct gaacgacacg cgctacatca gccggttctt cgcgaacttc atccgcgagc
2221 acctcaagtt cgcggaatcg gacgataagc agaaggtgta caccgtcaac ggccgtgtga
2281 cggcgcatct gcgctcgcgc tgggagttca acaagaaccg cgaagaatcg gatctgcacc
2341 acgccgtgga cgccgtgatc gtggcctgca cgaccccgtc ggacatcgcc aaggtgaccg
2401 ccttctatca gcgccgcgag cagaacaagg aactggccaa aaagaccgaa ccgcatttcc
2461 cgcagccgtg gccgcatttt gccgatgagc tgcgcgcgcg cctgtcgaag cacccgaaag
2521 aatcgatcaa ggccctgaac ctggggaact atgacgacca gaagctggag tccctgcagc
2581 ccgtgttcgt ctcccggatg cccaagcgca gcgtcacggg cgcggcccat caggaaacgc
2641 tgcgccgcta cgtgggcatc gacgaacgtt cggggaagat tcagacggtg gtgaaaacca
2701 agctgagcga aatcaagctc gatgcgtcgg gccactttcc catgtacggg aaggagagcg
2761 atccccgtac ctacgaggcg atccggcagc gcctgctgga acacaacaac gaccccaaga
2821 aagcctttca ggagcccctc tacaagccga agaagaacgg tgagccgggc ccggtcattc
2881 ggaccgtgaa gatcatcgac acgaagaacc aggtgattcc gctgaacgat gggaaaacgg
2941 tggcctacaa ctcgaatatc gtgcgggtgg acgtgttcga gaaggacggc aagtactact
3001 gcgtccccgt ctatacgatg gacatcatga agggcatcct gcccaataag gccattgagc
3061 cgaacaagcc gtacagcgaa tggaaggaga tgacggagga ctacacgttc cggttctcgc
3121 tctaccccaa cgatctgatc cgcatcgaac tgccgcgtga aaagatcatt aagacggccg
3181 gcggcgagga aatcaaaatc aaggatctct ttgcgtacta caaaaccatc cactcgggga
3241 ccgccggcct ggagctggtc agccacgact gctcgttcag cctgtccggc gtgggttcgc
3301 gcacgctgaa gcgctttgag aagtaccagg tggacgtgct gggcaacatc tacaaagtgc
3361 gcggcgagaa gcgggtgggc ctggcgagct cggcccacag caagaccggt gagacgattc
3421 gcccgctgca gtcgacgcgg gactagaagc ttggctgttt tggcggatga gagaagattt
3481 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
3541 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
3601 gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
3661 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
3721 gctctcctgc tagcgggagt ataacagaaa ccttaaggcc cgaccgcttg acaagggcgc
3781 gtgaggtttt tacgatagcg ccggatgcgg ggaaaaaggg ctccttttgg ggggttttcc
3841 ccgcaccggg cggacctggg cggagaggaa acgcggcaac tcgcccgtct cgggttcccg
3901 cccacgaccc ttaaggaggt gtgaggatta tgaatggacc aataataatg actagagaag
3961 aaagaatgaa gattgttcat gaaattaagg aacgaatatt ggataaatat ggggatgatg
4021 ttaaggctat tggtgtttat ggctctcttg gtcgtcagac tgatgggccc tattcggata
4081 ttgagatgat gtgtgtcatg tcaacagagg aagcagagtt cagccatgaa tggacaaccg
4141 gtgagtggaa ggtggaagtg aatttttata gcgaagagat tctactagat tatgcatctc
4201 aggtggaatc agattggccg cttacacatg gtcaattttt ctctattttg ccgatttatg
4261 attcaggtgg atacttagag aaagtgtatc aaactgctaa atcggtagaa gcccaaaagt
4321 tccacgatgc gatttgtgcc cttatcgtag aagagctgtt tgaatatgca ggcaaatggc
4381 gtaatattcg tgtgcaagga ccgacaacat ttctaccatc cttgacggta caggtagcaa
4441 tggcaggtgc catgttgatt ggtctgcatc atcgcatctg ttatacgacg agcgcttcgg
4501 tcttaactga agcagttaag caatcagatc ttccttcagg ttatgaccat ctgtgccagt
4561 tcgtaatgtc tggtcaactt tccgactctg agaaacttct ggaatcgcta gagaatttct
4621 ggaatgggat tcaggagtgg acagaacgac acggatatat agtggatgtg tcaaaacgca
4681 taccattttg acaattgctg cagactagat gggggttgcc agcgcctcca atttttcaag
4741 tgaaatctat aacgcctgtt ggcatgcctc agttgacccc attgaccctt ctctctggag
4801 gtggtagcta gaggggaatg gtcccaccct ggagccccac cccttgccaa ccctgggccc
4861 ggtataatgc cgggcaatgg gacgaccggg ccccctgggg gcccaaggcg gcagggccgc
4921 gtgaggacca aaaagagagg ctcccgctgg gagggaaccc aaattccagc aaccttttcc
4981 ttcgcctaag gttatcccct cccagcggga aaggcaatag gcgccaaggc ggcgcgcggg
5041 ggagtgccgt ctccgacgcg cgaaacggga ggggtttatg ccccaagaag gagttgaagg
5101 gaagagctat aaagaacagc cggaaggact caccgagcgg ggccgggcct tcctcgaggc
5161 cctggcccgg aggaagcggg cgaggggcga ggagcttccg cccctctacc tgaagctcct
5221 ggagggggcc gcgcccccgg agaagggggt ccatgagccc ccgcccgagg aggcgcctcc
5281 cccggaggac ctccgccgga agctccggga ggcgccgcct tccccggccc tccccaaccg
5341 acaggagctc tccgcctccc cgcccccgcc cgagatgagg cgggacgcct ggagcctcgc
5401 cgaccgcctc ctggaggagg gggagcggcg gggcaccctg ccagggcttt ccgagcggga
5461 gcggagggtg taccggaccc tcctcgccct gggcctcgag gtcctggccc ggaggctggg
5521 cccggggagg cccttgccca ggaacctgtc ccaggtctcc ttcttcgccg tgaacgacgc
5581 cctggccgtg gccctgggaa tccccccggc ctccctctac cgggtcctgg cctccctgga
5641 ggccaagggg ctcatccgcc ggagggcctg gcgcaccccg gccaccctca agggccggac
5701 gggggtctac gccggcggga ccctctacgc cgtgcgcctg ccccaccggg aggcccgccc
5761 ccgcctggac ccggaggact tccgccaccc ctggcgggac ctggaggggg acgcccgtca
5821 ggggcgcacc gcctggagct tgagagagtc atatacaagt cctcctaagg aggactccgg
5881 ggtcctccag ctcctccttc ggttttcgtt atcccctggc gaagccgaaa ctccgttagc
5941 tttagactct ctcaccgccc tcctccgggc ccgccccgcc gagcgccggg ccctggtgga
6001 ggccctcgcc ctctccctgg cccgggagtt ccgggacccc gggagcgtgc gcttctacgc
6061 ctgggtcctc tggaacgccc tccgggccga gctctacggc ctgatggagg gggccctcga
6121 ggccgtcgcc tgggcggtcc ggcgggcgcg ggaggccgcg gccaaggtcc tctggagccc
6181 gaggggcgag ggggtccggc gccccggggc cctcctcgcc cacctcctcc gggagcgggg
6241 cctcctggag ctcttccgcc aggcccctca gtggcgggtg gcgtagggct cccgccgggc
6301 taagctggag gtagcccatc cagggcctct ggggggtggc ttgtgtatca agactggctg
6361 ttcaaggact gcccaggggg ctggcgcctc cgctaccgcc ggaacggggg cggctacgag
6421 gcctcccggg acggtgagag gtgggagaag gtcccctcgg tcacggaggt tacggggcgc
6481 ctgaacaaga accttcagga ctgggccgtg cgccaggtgg tggagtacct ccagggggag
6541 ctggtcccgg gggtactgag ctgttgacaa ttaatcatcc ggctcgtata atgtgtggaa
6601 cgtcttctaa ggaggtcaca tatgaccatg attacggatt cactggccgt cgttttacaa
6661 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
6721 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
6781 agcctgaatg gcgaatagga agactagtca tagttcccct gagattatcg ctgtggtata
6841 atatcatacc acagcaatga tctcagggtt actatgataa gggctttctg cctaaggcag
6901 actgacccgc ggcgttgggg atcgcctgtc gcccgctttt ggcgggcatt ccccatcctt
6961 gcgcaaactc agaccttggc ggaaaacgct aaggtctttt ggtaccaccg aattccgagc
7021 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
7081 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
7141 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
7201 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
7261 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
7321 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
7381 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
7441 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
7501 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
7561 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
7621 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
7681 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
7741 tcctttgatc ttttctacgg ggtctgacgc tcagtgga
//
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