| 质粒名称: | pQCascade(Ptr)-entry(BsaI) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 175579 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pCDFDuet-1 |
| 完整质粒大小: | 8138 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | Streptomycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PtrTniQ, PtrCas5/8, PtrCas7, PtrCas6, CRISPR(BsaI) |
| 物种来源: | Pseudoalteromonas translucida |
| 基因突变: | No |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pQCascade(Ptr)-entry(BsaI) was a gift from Sheng Yang (Addgene plasmid # 175579 ; http://n2t.net/addgene:175579 ; RRID:Addgene_175579) |
| 参考文献部分: | Orthogonal CRISPR-associated transposases for parallel and multiplexed chromosomal integration. Yang S, Zhang Y, Xu J, Zhang J, Zhang J, Yang J, Jiang Y, Yang S. Nucleic Acids Res. 2021 Sep 3. pii: 6363765. doi: 10.1093/nar/gkab752. 10.1093/nar/gkab752 PubMed 34478496 |
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
-entry(BsaI)质粒图谱.png)
载体简介
载体序列
LOCUS pQCascade(Ptr)-entry(BsaI) 8138 bp ds-DNA circular SYN 26-OCT-2021
DEFINITION Expression of CRISPR-associated transposases.
ACCESSION .
VERSION .
KEYWORDS pQCascade(Ptr)-entry(BsaI)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8138)
AUTHORS Yang S, Zhang Y, Xu J, Zhang J, Zhang J, Yang J, Jiang Y, Yang S
TITLE Orthogonal CRISPR-associated transposases for parallel and
multiplexed chromosomal integration.
JOURNAL Nucleic Acids Res. 2021 Sep 3. pii: 6363765. doi:
10.1093/nar/gkab752.
PUBMED 34478496
REFERENCE 2 (bases 1 to 8138)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 26, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8138
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..56
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 7..25
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
protein_bind 37..55
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
promoter 263..318
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 269..287
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
protein_bind 299..317
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind complement(5305..5323)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5319..5366
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(5537..6328)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
promoter complement(6329..6420)
/gene="bla"
/label=AmpR promoter
CDS complement(7497..8120)
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind 8028..8047
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
ORIGIN
1 gttgacactc tatcattgat agagttattt taccactccc tatcagtgat agagaaaagt
61 gaactctaga aataattttg tttaacttta aaaggagata taccatgggt gaactgccga
121 gtaggcagct ggaaatgaga cctctggtct cgtgaactgc cgagtaggca gctgaagttg
181 gatccgaatt cgagctcggc gcgcctgcag gtcgacaagc ttgcggccgc tctaatctag
241 acatcattaa ttcctaattt ttgttgacac tctatcattg atagagttat tttaccactc
301 cctatcagtg atagagaaaa gtgaactcta gaaataattt tgtttaactt taaaggagat
361 atacatatgt ttttgcaaag acctaaataa ccaacaagaa ggagatatac atatgcactt
421 tctggtgcag accaagagct acccggacga ggcgctggaa agctatctgc tgcgtctggc
481 gcgtgataac agctacaacg gttatagcga gctggcggac atcctgtggc agtggctggc
541 ggaacaagat aacgagctgg aaggtgcgct gccgctggcg ctgagcaagg tggacgttta
601 ccacgcgcgt caggcgagca gcttccgtat ccgtgcgctg aaactggtgg cgcaactggc
661 ggacgttaac gcgggtgata ttctggcgct ggcgtggcgt cgtagcaact tcaagtttgg
721 caacctggcg gcggtgagcc gtaacgagct ggcgatcccg ctggaactgc tgcgtaccga
781 taacatcccg gtttgcatta aatgcctgag cgagagcagc cacattccgt tttactggca
841 cctgaagccg tataaagcgt gccacaagca caaaagccag ctgatcaccc gttgcaagga
901 gtgctacgac ctgattgatt atcgtgcgag cgaggcgttt ctggaatgcg tttgcggttg
961 caaaatcacc aacagcgaac aactgaacga cgcggatttc aagatcgcga ttgcgctggc
1021 gagcagcaac agccagaaaa tcgtgggcct gattagctgg ttcgcgaagg tgaaacaact
1081 ggacgttagc gacgcggatt tcaactgcgc gtttgttgat tacttcaaca cctggccgga
1141 gagcctgacc accgaactgg acctgctgac caacaacgcg cgtctgaagc agctgaaccc
1201 gtttaacaag accaaattca gcagcgtgta cggtgacctg atccgtgatg gccaaattgc
1261 ggcgaccagc aaccgtaaga acaaagttat cgacgagatc attagctatt ttgtggaact
1321 ggttgatagc aacccgaagg cgaaacaccc gaacattggt gacctgctgc tgtgcacctt
1381 cgatgcggcg gtgctgctga acaccaccac cgagcaggtt taccgtctgc accaagaagc
1441 gtttctgaac tgcgcgtata gccagaagaa acacgaacaa ctgcgtgcgg atagccacgt
1501 gttctatctg cgtcaggtta tcgagctgca gcaagcgttt gcggcggaaa aaccgctgac
1561 caagaaacaa ttcattgcgc cgtggtaact tatgaacctg caggatgcgc tggcgattga
1621 gccgctgaag gaaaaaacca ccgcgctgcg taagctgttc gtgccgtaca ccagccacgt
1681 tgaggtggat ggttttgagg aactggcgct gaccgtgctg atcaacctgg tttataagcg
1741 tagcgaaatt gacgatctga ccagcgcgcg taccgcgaaa agcgtgctgc gtgacgaggt
1801 tctgctgagc aagtgcatca acgaagtgaa atggttccac acccacaacc tgaagtaccc
1861 ggacatccgt gttagccacc aacgtctgat tagcgaggtg gttagcgaag atatcgcggg
1921 tatttgcagc cgtagcctgc cgctgagctt tggctggagc cacaacagcg cggagatcaa
1981 ccacgcgaaa ctgttcctga ccagctttaa ctggcagggt gaagtgacct gcctggcgcg
2041 tctgctgatt aacgaggaac cggtttggat caacctgatt cgtgcgtacg gtttcaccaa
2101 gaaagcggtt ctggagatca gcggcaagat taaacagcaa ctgccggtgg cggagttccc
2161 gctggaagtt agcagcttta gcccgcagct gcaaatgccg tttcagcaaa gctatctggt
2221 ggttaccccg gtggttagcc acgcgatgct ggcgaagatc cagcaactga ccaccgaccg
2281 taaactgaac ttcgcgctgg ttgagcacag ccgtccggcg aacgttggtg atctggcgag
2341 cagcgtgggt ggcaacattc gtgttctgcg ttactttccg aagacctata gcaaagcggt
2401 gaaccgtagc aaagttgcga acaacgatat cgaaaaggcg ttcaaaattc gtgcgctgct
2461 gagcagccag tttcagcaag cgctgctggt gctggttggc atcaagcagt tcaacaccct
2521 gcgtcaaaaa cgtctggcgc gtgtggcggc gatccgtcaa gtgcgtgtta gcctgcaact
2581 gtggctggac aacattctgg aggcgaagaa caacgcgcag aaccaagtgt acccggaatg
2641 ggttcgtcac tatctggatc aaagcatcac caactgcatt agccagttca gcaacgttct
2701 gaacgaaagc ctgggtaacc tgagcaagct gaaacgtttt gcgtaccacc cgaacctgat
2761 gggcctgttc aaagcgcaac tgaactatgt gtttacccac tgcgcggcgg agcaggaaat
2821 cctgaacgac gagcaaattg tgtacgttca ctgccaggac atgcgtgttt tcgatgcgga
2881 agcgatggcg aacccgtata tccagggtat gccgagcctg accgcgctga acggcctggc
2941 gcacaacttc gagcgtaagc tgaaaaactt tattgatccg agcatcaagt gcattggtag
3001 cgcgatctac attgagaact atcaactgca caccggcaaa ccgctgccgg aaccgagcaa
3061 gctgaaacag gtggcgggtc gtagccacgt tatccgtagc ggcatcattg acaagccgaa
3121 atgcgacatt accctggatc tggtgttccg tctgtttgtt ccgaacaccg aactgctgga
3181 taagctgaac agccaactga ttaagccggc gctgccgagc agctttgcgg gtggcaccat
3241 gcacccgccg agcctgtacc agaacattga ctggtgccac gtgcacacca agccgagcga
3301 gctgtttaag aaactgaagg cgaaaagcag caacggtagc tggctgtatc cgagcaagaa
3361 agtggttaaa agcttcgaac agctgatcga cgcgctgaac agcaacttta acctgcgtcc
3421 ggcggcgatt ggcctggcgg cgctggagga accggtgaag cgtgatgcgg cgctgcacga
3481 gtaccactgc tatgcggaac cggttatcgg tctgctggag tgcgtgagca acaccagcgt
3541 taagtacgcg ggcgcgaaac aattctttca cgacgcgttc tgggtgatgg atgttcagaa
3601 ggaaagcatg ctgatgaaga aaagcaaatt tgagtatgaa taatgcagct gccgcgtcac
3661 ctgagctaca cccgtagcct gagcccgagc aaggcggtgt tcttttataa aaccccggag
3721 agcgacttcg aaccgctgca gatcgagcaa aacaaactgg tgggtcagaa gagcggtttt
3781 ggcgatgcgt accagaagca aaacgttgcg aaaaacctgg cgccgcagga cctggcgttt
3841 ggtaacccgc aaaccattga tgtgtgctat gttccgccga ccgtgaacga actgttctgc
3901 cgttttagcc tgcgtgttga ggcgaactgc atcgaaccgc acgtgtgcga cgatccgaag
3961 gttatttact ggctgaaacg tttctttgaa acctataaga aacacaacgg tctgaacgaa
4021 gtggcgaccc gttacgcgaa gaacatcctg atgggcaact ggctgtggcg taaccgtcag
4081 agcccgaacg ttgacatcga gattctgacc gaacacgcgg cgccgattgt ggttgagggt
4141 gcgcagaagc tgaaatggca aggcaactgg cagaacaacc aaaccgcgct gctgaccctg
4201 agcgagagca tccaggaagg tctgagcaac ccgcaaaact actgctatct ggatatcacc
4261 gcgaagatta aaaacgcgtt cagccaggaa gtgcacccga gccaaaagtt tgtggacaac
4321 gttgaacagg gtatgagcag caaacagctg gcgtataccc aagtgggcga taagaaagcg
4381 gcgagcctga acagccagaa ggttggcgcg gcgatccaaa ccattgacga ttggtacgag
4441 gaaggttata aaccgctgcg tacccatgag tatggtgcgg acaagcaaat cctggtggcg
4501 caccgtaccc cgaaaagcca cagcgatttt tatagcctgc tgccgcgtat cgcgctgcac
4561 attaagcaca tggaaaaaca cggtctggag cagagcgaac aaagcaacag catccacttc
4621 attgcggcgg ttctgattaa gggtggcctg tttcagcgta gcaaaggatg aagcgttact
4681 atttcaccat cacctacctg ccgcaaagct gcgatgtgag cctgctggcg ggtcgttgca
4741 tcggcattct gcacggtttc atgagcagcc gtgagatcag caacattggc gtgtgctttc
4801 cgaaatggaa cgagcagacc atcggtaacg aactggcgtt tgttagcacc aacaagaaac
4861 aactgaccaa cctgagccag caaagctatt tcgagatgat ggcgcacgac aagctgtttg
4921 gcctgagcaa aattctggaa gtgccggtta accagagcga agtgatgttc gttcgtaacc
4981 aaagcgtggc gaaggcgttt gttggtgaaa agcaacgtcg tctgaaacgt gcgaagaaac
5041 gtgcggaggc gcgtggcgaa gtgtacaacc cggagtataa gttcgaagcg aaagatatcg
5101 gtcactttca cagcattccg gtgagcagca agggtaacgg ccagagctac gttctgcaca
5161 tccaaaagaa cgagaacgcg gaaagcatta aaaaccagtt caacaactat ggctttgcga
5221 ccaaccaaat tttcctgggc accgtgccga gcctgaacac cctgctgtaa ggtaccaccc
5281 ttaatctgac ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct
5341 ctaaacgggt cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca
5401 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
5461 gaccctgccc tgaaccgacg accgggtcat cgtggccgga tcttgcggcc cctcggcttg
5521 aacgaattgt tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga
5581 caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag
5641 cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt
5701 cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca
5761 acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta
5821 aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg
5881 ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat
5941 caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc
6001 caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa
6061 tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa
6121 ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca
6181 aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg
6241 ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg
6301 atacttcggc gatcaccgct tccctcatac tcttcctttt tcaatattat tgaagcattt
6361 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
6421 tagctagctc actcggtcgc tacgctccgg gcgtgagact gcggcgggcg ctgcggacac
6481 atacaaagtt acccacagat tccgtggata agcaggggac taacatgtga ggcaaaacag
6541 cagggccgcg ccggtggcgt ttttccatag gctccgccct cctgccagag ttcacataaa
6601 cagacgcttt tccggtgcat ctgtgggagc cgtgaggctc aaccatgaat ctgacagtac
6661 gggcgaaacc cgacaggact taaagatccc caccgtttcc ggcgggtcgc tccctcttgc
6721 gctctcctgt tccgaccctg ccgtttaccg gatacctgtt ccgcctttct cccttacggg
6781 aagtgtggcg ctttctcata gctcacacac tggtatctcg gctcggtgta ggtcgttcgc
6841 tccaagctgg gctgtaagca agaactcccc gttcagcccg actgctgcgc cttatccggt
6901 aactgttcac ttgagtccaa cccggaaaag cacggtaaaa cgccactggc agcagccatt
6961 ggtaactggg agttcgcaga ggatttgttt agctaaacac gcggttgctc ttgaagtgtg
7021 cgccaaagtc cggctacact ggaaggacag atttggttgc tgtgctctgc gaaagccagt
7081 taccacggtt aagcagttcc ccaactgact taaccttcga tcaaaccacc tccccaggtg
7141 gttttttcga aggttaagtc agttggggaa ctgcttaacc ttcgatcaaa ccacctcccc
7201 aggtggtttt ttcgtttaca gggcaaaaga ttacgcgcag aaaaaaagga tctcaagaag
7261 atcctttgat cttttctact gaaccgctct agatttcagt gcaatttatc tcttcaaatg
7321 tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag tcatgccccg
7381 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcctga tgaatcccct
7441 aatgattttt atcaaaatca ttaaggttac catcacggaa aaaggttatg ctgcttttaa
7501 gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc aaggccgaat
7561 aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg taataatggc
7621 ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg ctcttgatct
7681 tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa tgcattctct
7741 agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc atactgtttt
7801 tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag taaagcacat
7861 ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc taaagggcaa
7921 aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa agcccgctta
7981 ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc gagtttacgg
8041 gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt aatcacttta
8101 cttttatcta atctagacat cattaattcc taattttt
//
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