| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pCAMBIA0305.1 |
| 质粒类型: | 植物载体；农杆菌双元表达载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | 限制性内切酶，多克隆位点 |
| 启动子: | lac |
| 载体大小: | 9662 bp |
| 5' 测序引物及序列: | M13 Reverse:CAGGAAACAGCTATGAC |
| 3' 测序引物及序列: | M13/pUC Forward:CCCAGTCACGACGTTGTAAAACG |
| 载体标签: | GUS、6xHis |
| 载体抗性: | 卡那霉素 |
| 筛选标记: | -- |
| 克隆菌株: | HB101等菌株 |
| 宿主细胞（系）: | 植物细胞、农杆菌 |
| 备注: | -- |
| 产品目录号: | -- |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 诱导型 |
| 病毒/非病毒: | 非病毒 |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pCAMBIA0305.1
- 库存：
20
- 供应商：
泽叶生物
载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pCAMBIA0305.1 9662 bp ds-DNA circular SYN 25-JUL-2016
DEFINITION Agrobacterium binary vector for plant transformation, with
kanamycin-resistance and GUSPlus(TM) genes.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9662)
AUTHORS Cambia
TITLE Direct Submission
JOURNAL Exported Monday, July 25, 2016 from SnapGene Viewer 3.1.4 to Vector
NTI(R) format
http://www.snapgene.com
COMMENT The sequence was corrected by inserting a G at position 4740.
COMMENT VNTDATE|658800000|
COMMENT VNTDBDATE|776304000|
COMMENT VNTNAME|pCAMBIA0305.1|
COMMENT VNTAUTHORNAME|Cambia|
FEATURES Location/Qualifiers
CDS join(2..16,207..2024)
/codon_start=1
/product="beta-glucuronidase"
/label=GUSPlus(TM)
/note="codon-optimized Staphylococcus gusA gene with a
catalase intron to ensure expression in plants but not
bacteria"
intron 17..206
/label=cat1 intron
/note="castor bean catalase intron, modified"
CDS 2031..2048
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
terminator 2080..2332
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 2354..2378
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 3678..4307
/codon_start=1
/product="stability protein from Pseudomonas plasmid pVS1"
/label=pVS1 StaA
CDS 4736..5809
/codon_start=1
/product="replication protein from Pseudomonas plasmid
pVS1"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 5875..6069
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 6413..6553
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(6739..7327)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7414..8208)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
misc_feature 8633..8657
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
promoter 8761..8791
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8799..8815
/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)."
primer_bind 8823..8839
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 8835..9068
/codon_start=1
/gene="lacZ (truncated)"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
misc_feature 8849..8905
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(8909..8925)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 9302..9647
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus
CaMV 35S promoter"
ORIGIN
1 catggtagat ctgagggtaa atttctagtt tttctccttc attttcttgg ttaggaccct
61 tttctctttt tatttttttg agctttgatc tttctttaaa ctgatctatt ttttaattga
121 ttggttatgg tgtaaatatt acatagcttt aactgataat ctgattactt tatttcgtgt
181 gtctatgatg atgatgatag ttacagaacc gacgaactag tctgtacccg atcaacaccg
241 agacccgtgg cgtcttcgac ctcaatggcg tctggaactt caagctggac tacgggaaag
301 gactggaaga gaagtggtac gaaagcaagc tgaccgacac tattagtatg gccgtcccaa
361 gcagttacaa tgacattggc gtgaccaagg aaatccgcaa ccatatcgga tatgtctggt
421 acgaacgtga gttcacggtg ccggcctatc tgaaggatca gcgtatcgtg ctccgcttcg
481 gctctgcaac tcacaaagca attgtctatg tcaatggtga gctggtcgtg gagcacaagg
541 gcggattcct gccattcgaa gcggaaatca acaactcgct gcgtgatggc atgaatcgcg
601 tcaccgtcgc cgtggacaac atcctcgacg atagcaccct cccggtgggg ctgtacagcg
661 agcgccacga agagggcctc ggaaaagtca ttcgtaacaa gccgaacttc gacttcttca
721 actatgcagg cctgcaccgt ccggtgaaaa tctacacgac cccgtttacg tacgtcgagg
781 acatctcggt tgtgaccgac ttcaatggcc caaccgggac tgtgacctat acggtggact
841 ttcaaggcaa agccgagacc gtgaaagtgt cggtcgtgga tgaggaaggc aaagtggtcg
901 caagcaccga gggcctgagc ggtaacgtgg agattccgaa tgtcatcctc tgggaaccac
961 tgaacacgta tctctaccag atcaaagtgg aactggtgaa cgacggactg accatcgatg
1021 tctatgaaga gccgttcggc gtgcggaccg tggaagtcaa cgacggcaag ttcctcatca
1081 acaacaaacc gttctacttc aagggctttg gcaaacatga ggacactcct atcaacggcc
1141 gtggctttaa cgaagcgagc aatgtgatgg atttcaatat cctcaaatgg atcggcgcca
1201 acagcttccg gaccgcacac tatccgtact ctgaagagtt gatgcgtctt gcggatcgcg
1261 agggtctggt cgtgatcgac gagactccgg cagttggcgt gcacctcaac ttcatggcca
1321 ccacgggact cggcgaaggc agcgagcgcg tcagtacctg ggagaagatt cggacgtttg
1381 agcaccatca agacgttctc cgtgaactgg tgtctcgtga caagaaccat ccaagcgtcg
1441 tgatgtggag catcgccaac gaggcggcga ctgaggaaga gggcgcgtac gagtacttca
1501 agccgttggt ggagctgacc aaggaactcg acccacagaa gcgtccggtc acgatcgtgc
1561 tgtttgtgat ggctaccccg gagacggaca aagtcgccga actgattgac gtcatcgcgc
1621 tcaatcgcta taacggatgg tacttcgatg gcggtgatct cgaagcggcc aaagtccatc
1681 tccgccagga atttcacgcg tggaacaagc gttgcccagg aaagccgatc atgatcactg
1741 agtacggcgc agacaccgtt gcgggctttc acgacattga tccagtgatg ttcaccgagg
1801 aatatcaagt cgagtactac caggcgaacc acgtcgtgtt cgatgagttt gagaacttcg
1861 tgggtgagca agcgtggaac ttcgcggact tcgcgacctc tcagggcgtg atgcgcgtcc
1921 aaggaaacaa gaagggcgtg ttcactcgtg accgcaagcc gaagctcgcc gcgcacgtct
1981 ttcgcgagcg ctggaccaac attccagatt tcggctacaa gaacgctagc catcaccatc
2041 accatcacgt gtgaattggt gaccagctcg aatttccccg atcgttcaaa catttggcaa
2101 taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg
2161 ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg
2221 gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag
2281 cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcgggaatta
2341 aactatcagt gtttgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt
2401 agaataacgg atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc
2461 atgccaacca cagggttccc ctcgggatca aagtactttg atccaacccc tccgctgcta
2521 tagtgcagtc ggcttctgac gttcagtgca gccgtcttct gaaaacgaca tgtcgcacaa
2581 gtcctaagtt acgcgacagg ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg
2641 ttttagtcgc ataaagtaga atacttgcga ctagaaccgg agacattacg ccatgaacaa
2701 gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc accgacgacc aggacttgac
2761 caaccaacgg gccgaactgc acgcggccgg ctgcaccaag ctgttttccg agaagatcac
2821 cggcaccagg cgcgaccgcc cggagctggc caggatgctt gaccacctac gccctggcga
2881 cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc acccgcgacc tactggacat
2941 tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga
3001 caccaccacg ccggccggcc gcatggtgtt gaccgtgttc gccggcattg ccgagttcga
3061 gcgttcccta atcatcgacc gcacccggag cgggcgcgag gccgccaagg cccgaggcgt
3121 gaagtttggc ccccgcccta ccctcacccc ggcacagatc gcgcacgccc gcgagctgat
3181 cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac
3241 cctgtaccgc gcacttgagc gcagcgagga agtgacgccc accgaggcca ggcggcgcgg
3301 tgccttccgt gaggacgcat tgaccgaggc cgacgccctg gcggccgccg agaatgaacg
3361 ccaagaggaa caagcatgaa accgcaccag gacggccagg acgaaccgtt tttcattacc
3421 gaagagatcg aggcggagat gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacgtc
3481 tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg atgccaagct ggcggcctgg
3541 ccggccagct tggccgctga agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt
3601 gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa
3661 atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca
3721 agacgaccat cgcaacccat ctagcccgcg ccctgcaact cgccggggcc gatgttctgt
3781 tagtcgattc cgatccccag ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac
3841 cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg cgacgtgaag gccatcggcc
3901 ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc ggacttggct gtgtccgcga
3961 tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag cccttacgac atatgggcca
4021 ccgccgacct ggtggagctg gttaagcagc gcattgaggt cacggatgga aggctacaag
4081 cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg
4141 cgctggccgg gtacgagctg cccattcttg agtcccgtat cacgcagcgc gtgagctacc
4201 caggcactgc cgccgccggc acaaccgttc ttgaatcaga acccgagggc gacgctgccc
4261 gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact catttgagtt aatgaggtaa
4321 agagaaaatg agcaaaagca caaacacgct aagtgccggc cgtccgagcg cacgcagcag
4381 caaggctgca acgttggcca gcctggcaga cacgccagcc atgaagcggg tcaactttca
4441 gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat
4501 taccgagctg ctatctgaat acatcgcgca gctaccagag taaatgagca aatgaataaa
4561 tgagtagatg aattttagcg gctaaaggag gcggcatgga aaatcaagaa caaccaggca
4621 ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc gtaagcggct
4681 gggttgtctg ccggccctgc aatggcactg gaacccccaa gcccgaggaa tcggcgtgag
4741 cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga
4801 gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg
4861 tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc
4921 cggtgcgccg tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc
4981 gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg
5041 tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca
5101 cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact
5161 gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa
5221 gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga
5281 tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt
5341 tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga
5401 agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga
5461 gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct
5521 gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct
5581 ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg
5641 cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc
5701 aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt
5761 catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca
5821 gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaggtctct ttcctgtgga
5881 tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa
5941 cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa
6001 aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc
6061 ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg
6121 gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc
6181 aaaaatggct ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc
6241 actcgaccgc cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg
6301 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
6361 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
6421 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
6481 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
6541 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
6601 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
6661 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
6721 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
6781 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
6841 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
6901 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
6961 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
7021 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
7081 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
7141 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
7201 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
7261 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
7321 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
7381 acgttaaggg attttggtca tgcattctag gtactaaaac aattcatcca gtaaaatata
7441 atattttatt ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata
7501 ctgttcttcc ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt
7561 gtccgccctg ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa
7621 gatgttgctg tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt
7681 ctttaaaaaa tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc
7741 gcaatccaca tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc
7801 taagctattc gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc
7861 cgcatacagc tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag
7921 gacgccatcg gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag
7981 gacctttgga acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac
8041 atcataggtg gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc
8101 tcccaccagc ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta
8161 tttttcgatc agttttttca attccggtga tattctcatt ttagccattt attatttcct
8221 tcctcttttc tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc
8281 aattcactgt tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg
8341 ttttcaaagt tggcgtataa catagtatcg acggagccga ttttgaaacc gcggtgatca
8401 caggcagcaa cgctctgtca tcgttacaat caacatgcta ccctccgcga gatcatccgt
8461 gtttcaaacc cggcagctta gttgccgttc ttccgaatag catcggtaac atgagcaaag
8521 tctgccgcct tacaacggct ctcccgctga cgccgtcccg gactgatggg ctgcctgtat
8581 cgagtggtga ttttgtgccg agctgccggt cggggagctg ttggctggct ggtggcagga
8641 tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa taacacattg cggacgtttt
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1.常用载体分类：2,常用载体元件介绍：(1) 启动子：启动子是位于结构基因5'端上游的DNA序列，能活化RNA聚合酶，使之与模板DNA准确的结合并具有转录起始的特异性。启动子主要分为广谱启动子和特异性启动子。广谱启动子为在各种组织、细胞上广泛表达的启动子；特异性启动子为在特异性组织或者细胞类型上表达的启动子。常用广谱启动子有CAG，CMV，EF1a，PGK等，还包含启动RNA的III型启动子U6和H1启动子；常用特异性启动子如成熟神经元特异性启动子hSyn，投射神经元特异性启动
恰当的选择固相载体和连接分子是固相反应的重要组成部分. 固相载体和连接分子都应该在多种遗传反应过程期间是保持稳定, 而且产品的切除应该是在温和的条件下实现. 本章将介绍各种不同的载体和连接分子. 如果反应后可以用过滤的方法去除, 从理论上讲, 任何材料都可用作固相载体. 聚乙烯, 聚苯乙烯-外涂聚乙烯薄层, 平滑的纸或棉花等都可以. 最常用的载体是: 聚苯乙烯(PS), 聚乙二醇(PEG), Tentagel (TG), 和PEGA (聚乙二醇-聚丙
zhangyuxianggx 各位大侠好！现有pcDNA3-mCherry和pGEM-hgene两个东西在滤纸上，老师就说让构建载体，具体是什么意思啊？我理解的是一个真核表达载体和构建好的pGEM质粒，但是不知下一步该怎么做？恳请各位好心的大侠指点一二。 shylook 直接插入cDNA吧 zhangyuxianggx 那个pGEM就是已经插入了DNA的载体。请问这两个东西溶下来以后
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pCAMBIA0305.1载体
¥1800
