| 出品公司: | Invitrogen |
|---|---|
| 载体名称: | pDONR222 |
| 质粒类型: | Gateway载体；克隆载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 启动子: | 无 |
| 载体大小: | 4718bp |
| 5' 测序引物及序列: | M13/pUC fwd 5'CCCAGTCACGACGTTGTAAAACG 3' |
| 3' 测序引物及序列: | M13/pUC rev 5' AGCGGATAACAATTTCACACAGG 3' |
| 载体标签: | 无 |
| 载体抗性: | 卡那霉素，氯霉素（仅空载体） |
| 真核筛选标记: | 无 |
| 克隆菌株: | 阳性筛选用DB3.1 ，阴性筛选用OmniMAX2-T1 |
| 备注: | pDONR222载体在用来建入门克隆。 |
| 产品目录号: | -- |
| 病毒/非病毒: | 非病毒 |
买家导航
pDONR222载体质粒基本信息
pDONR222质粒图谱载体图谱和pDONR222载体序列质粒序列多克隆位点信息
pDONR222质粒载体简介
pDONR222质粒序列载体序列
LOCUS Exported 4718 bp ds-DNA circular SYN 09-AUG-2016
DEFINITION synthetic circular DNA
KEYWORDS pDONR222
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4718)
AUTHORS Biofeng Lab.
FEATURES Location/Qualifiers
source 1..4718
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 58..85
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 177..263
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 327..343
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 360..591
/gene="mutant version of attP"
/bound_moiety="BP Clonase(TM)"
/note="attP1"
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)201 version)"
CDS complement(987..1292)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/note="ccdB"
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(1642..2295)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
promoter complement(2296..2398)
/note="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
protein_bind complement(2543..2774)
/gene="mutant version of attP"
/bound_moiety="BP Clonase(TM)"
/note="attP2"
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)201 version)"
promoter complement(2793..2811)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2816..2832)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(2899..3714)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
promoter 3722..3777
/gene="tetR"
/note="tetR/tetA promoters"
/note="overlapping promoters for bacterial tetR and tetA"
rep_origin complement(4104..4692)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 gtctgacgct cagggaacga cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
61 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
121 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
181 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
241 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
301 atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca
361 aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg
421 cttttttata atgccaactt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa
481 tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca
541 acatatccag tcactatgaa tcaactactt agatggtatt agtgacctgt agtcgaccga
601 cagccttcca aatgttcttc gggtgatgct gccaacttag tcgaccgaca gccttccaaa
661 tgttcttctc aaacggaatc gtcgtatcca gcctactcgc tattgtcctc aatgccgtat
721 taaatcataa aaagaaataa ggaaaagagg tgcgagcctc ttttttgtgt gacaaaataa
781 aaacatctac ctattcatat acgctagtgt catagtcctg aaaatcatct gcatcaagaa
841 caatttcaca actcttatac ttttctctta caagtcgttc ggcttcatct ggattttcag
901 cctctatact tactaaacgt gataaagttt ctgtaatttc tactgtatcg acctgcagac
961 tggctgtgta taagggagcc tgacatttat attccccaga acatcaggtt aatggcgttt
1021 ttgatgtcat tttcgcggtg gctgagatca gccacttctt ccccgataac ggagaccggc
1081 acactggcca tatcggtggt catcatgcgc cagctttcat ccccgatatg caccaccggg
1141 taaagttcac gggagacttt atctgacagc agacgtgcac tggccagggg gatcaccatc
1201 cgtcgcccgg gcgtgtcaat aatatcactc tgtacatcca caaacagacg ataacggctc
1261 tctcttttat aggtgtaaac cttaaactgc atttcaccag cccctgttct cgtcagcaaa
1321 agagccgttc atttcaataa accgggcgac ctcagccatc ccttcctgat tttccgcttt
1381 ccagcgttcg gcacgcagac gacgggcttc attctgcatg gttgtgctta ccagaccgga
1441 gatattgaca tcatatatgc cttgagcaac tgatagctgt cgctgtcaac tgtcactgta
1501 atacgctgct tcatagcata cctctttttg acatacttcg ggtatacata tcagtatata
1561 ttcttatacc gcaaaaatca gcgcgcaaat acgcatactg ttatctggct tttagtaagc
1621 cggatccacg cggcgtttac gccccccctg ccactcatcg cagtactgtt gtaattcatt
1681 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
1741 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa
1801 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
1861 gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
1921 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
1981 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
2041 ccatatcacc agctcaccgt ctttcattgc catacggaat tccggatgag cattcatcag
2101 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
2161 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
2221 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
2281 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
2341 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac
2401 gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt
2461 tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg
2521 tgatgctgcc aacttagtcg actacaggtc actaatacca tctaagtagt tgattcatag
2581 tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt
2641 taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt acaaagttgg
2701 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
2761 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
2821 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
2881 taaaaatata tcatcatgtt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
2941 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
3001 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
3061 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
3121 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
3181 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
3241 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
3301 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
3361 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
3421 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
3481 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
3541 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
3601 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
3661 gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc tcatagatct
3721 tttctccatc actgataggg agtggtaaaa taactccatc aatgatagag tgtcaacaac
3781 atgaccaaaa tcccttaacg tgagttacgc gtattaattg cgttgcgctc actgcccgct
3841 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
3901 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
3961 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
4021 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
4081 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
4141 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
4201 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
4261 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
4321 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
4381 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
4441 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
4501 acagagttct tgaagtggtg gcctaactac ggttacacta gaagaacagt atttggtatc
4561 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
4621 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
4681 aaaggatctc aagaagatcc tttgatcttt tctacggg
//
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pDONR222质粒载体应用举例
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