| 质粒名称: | pDonorGFP(Ptr) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 175578 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pACYC |
| 完整质粒大小: | 4606 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | Chloramphenicol |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Mini-Tn, GFP cassette |
| 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: | pDonorGFP(Ptr) was a gift from Sheng Yang (Addgene plasmid # 175578 ; http://n2t.net/addgene:175578 ; RRID:Addgene_175578) |
| 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
pDonorGFP(Ptr)载体质粒基本信息
pDonorGFP(Ptr)质粒图谱载体图谱和pDonorGFP(Ptr)载体序列质粒序列多克隆位点信息
质粒图谱.png)
pDonorGFP(Ptr)质粒载体简介
pDonorGFP(Ptr)质粒序列载体序列
LOCUS pDonorGFP(Ptr) 4606 bp ds-DNA circular SYN 26-OCT-2021
DEFINITION DNA donor for transposition.
ACCESSION .
VERSION .
KEYWORDS pDonorGFP(Ptr)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4606)
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 4606)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 26, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4606
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(288..305)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(422..967)
/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(502..521)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
RBS 1349..1371
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1388..2095
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
CTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNY
KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKANF
KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
TAAGITHGMDELYK"
primer_bind complement(1412..1440)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 2035..2055
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
terminator 2189..2237
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
terminator 2299..2347
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
primer_bind complement(2416..2438)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3290..3309)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
CDS complement(3740..4399)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 4314..4333
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(4400..4502)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
ORIGIN
1 ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga tggtgttttt
61 gaggtgctcc agtggcttct gtttctatca gctgtccctc ctgttcagct actgacgggg
121 tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta tactggctta
181 ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct
241 gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac
301 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
361 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
421 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
481 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
541 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
601 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
661 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
721 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
781 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
841 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
901 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
961 atctcaagaa gatcatctta ttaatatacg tagtatctac gatacgtagt atctacgata
1021 cgtaggaggt tgagaagtca tttaataagg ccactgttaa acgtagatat atctacgcgt
1081 agatatatct acgatacgta gcggccgacg cgctgggcta cgtcttaatg ggcagcgccc
1141 acatacgcag cgatttccaa cagcttgagg cggaagcttt aatgcgggtc ctttgggtgt
1201 tgtttgaagt ataagttgac atatctgtac taaaagatgg cataaattgg aagtgtaagg
1261 tggcatagtc tagtatttaa ccaaatggtt aaatggttga ctcaccggga attccatggt
1321 cgacatgcat aagcttccta ggaataattt tgtttaactt taagaaggag atatacatat
1381 ggctagcaaa ggagaagaac tcttcactgg agttgtccca attcttgttg aattagatgg
1441 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
1501 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
1561 tgtcactact ttcacttatg gtgttcagtg cttttcaaga tacccggatc atatgaaacg
1621 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt
1681 caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt
1741 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttgggcacaa
1801 attggaatac aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg
1861 aatcaaagct aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga
1921 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
1981 cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct
2041 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaaggatc
2101 cggctgctaa tcgagttaat taaactagtg agctcggtac ccggggatcc tctagaggtc
2161 gaaattcacc tcgaaagcaa gctgataaac cgatacaatt aaaggctcct tttggagcct
2221 ttttttttgg agattttcaa cgtgaaaaaa ttattattcg caattccaag ctaattcacc
2281 tcgaaagcaa gctgataaac cgatacaatt aaaggctcct tttggagcct ttttttttgg
2341 agattttcaa cgtgaaaaaa ttattattcg caattccaag ctctgcctcg cgcgtttcgg
2401 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
2461 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
2521 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatgggctcg atcccctcgg
2581 atccaattgg tcttaatttt ccttaattat ttttaaagtt agactgattt agacttggaa
2641 aagcttaatg attggagagc taaattgact aatagtattc agtcaagttt aattagtttt
2701 aagcgatcca tgcataacta tttctgtaca atgctatatt ttcaccaatt aataattttt
2761 aatcaagcct acattatgaa atatactata cccatttgaa ctcttctatt tgtaccattg
2821 tcggtagcaa aaacttatgg ggttttgaac gtcacttaaa ttgtaagcat ttgcgatgga
2881 ggcgcgttta gagtcaacct tgattctgat atgctccgaa tttttggtaa gaatataagt
2941 gtgagagtag ctaatgtgga tacgcctgag ttaagggaaa aatgtgaaaa tgaaataact
3001 cgttatcatg caaagtgact aaggttataa tcttccgttt atggcacata gcagccaact
3061 aaacttgaca gtatttttat gtggttggct ttataaaacc agcatttggt aacattatgc
3121 caatttttac ttcaatatta tgccaacata cactacacta acggagctgt agcacaataa
3181 gctcgtttgt acttatgcca acttatactt caaacaacat tggggtggat cctctacgcc
3241 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
3301 gacatcaccg atggggaaga tcgggcctgc caccataccc acgccgaaac aagcgccctg
3361 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac
3421 atctgtatta acgaagcgct aaccgttttt atcaggctct gggaggcaga ataaatgatc
3481 atatcgtcaa ttattacctc cacggggaga gcctgagcaa actggcctca ggcatttgag
3541 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa
3601 gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc
3661 tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc
3721 aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
3781 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
3841 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
3901 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
3961 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
4021 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
4081 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
4141 tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca
4201 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
4261 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
4321 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
4381 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
4441 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
4501 caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag
4561 gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattc
//
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