| 质粒名称: | pRAD1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 47697 |
| 存储实验室: | Mary Lidstrom |
|
载体骨架基本信息 |
|
| 载体名称: | pI3 and pMTL23 |
| 载体出品公司: | - |
| 空载体大小: | 6280 bp |
| 完整质粒大小: | -- |
| 载体修饰: | pMTL23携带pI 12的SalI-FspI片段；6.3kb。 |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素 |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | repU |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | Cat-F |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pRAD1 was a gift from Mary Lidstrom (Addgene plasmid # 47697 ; http://n2t.net/addgene:47697 ; RRID:Addgene_47697) |
| 参考文献部分: | Characterization of the minimal replicon of a cryptic Deinococcus radiodurans SARK plasmid and development of versatile Escherichia coli-D. radiodurans shuttle vectors. Meima R, Lidstrom ME. Appl Environ Microbiol. 2000 Sep;66(9):3856-67. 10.1128/AEM.66.9.3856-3867.2000 PubMed 10966401 |
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 pRAD1 6809 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION General cloning vector for use in E. coli or D. radiodurans.
ACCESSION .
VERSION .
KEYWORDS pRAD1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6809)
AUTHORS Meima R, Lidstrom ME
TITLE Characterization of the minimal replicon of a cryptic Deinococcus
radiodurans SARK plasmid and development of versatile Escherichia
coli-D. radiodurans shuttle vectors.
JOURNAL Appl Environ Microbiol. 2000 Sep;66(9):3856-67.
PUBMED 10966401
REFERENCE 2 (bases 1 to 6809)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6809
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 27..131
/gene="bla"
/label=AmpR promoter
CDS 132..992
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(350..369)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1163..1751
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1652..1671
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1905..1922
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2039..2060
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2075..2105
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2113..2129
/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 2118..2140
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2137..2153
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2137..2153
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 5964..6623
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(6030..6049)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
2161 acgccaagct cgcgaggcct cgagatctat cgatgcatgc catggtaccc gggagctcga
2221 attctagaag cttctgcaga cgcgtcgacg tcatatggat ccgattcgac ctgcaggcat
2281 gcaagctgta cgcctcagct accattcttg cagccgcgtt cagggtcttg acgtgttctt
2341 tgtatctatc agcaaactga gaacccaatt ctggatcatc cttgtaacgc caactctccg
2401 accattccga aggaacgcct tgtagtcctc ctgagccgtt agggtatgtc tttagtccaa
2461 tcaaatggga catttgcaga caggaacgca caagtgtctc acgtcgcttc tctatttcaa
2521 tatctaaaaa ttcttcactc ttatcgttgc ttgacatatt aaaactgtct aactgatcca
2581 gttcacttat atggaaagta tcactaaagt catggcttct cagaaattca atacttccca
2641 ttgatgggga ccggttaacc aaatcgataa aaatttttct gtcatgtttc agataagttt
2701 ttggcgcgtt gattttataa taaagccgcc attcttgtgc caacaacgcc acgaaaagag
2761 ccacagcagc agcggccaca tcccattgaa aatctttcgc cagaccaacc gccaggagaa
2821 ccaagccgaa atagttagcc cagcgaacta aaaaactatc tataaacgta ttaagaaacg
2881 ttcgggtcat atttccgtga atcatatcag gacgtaaatt tgcacatgac gcatttccgg
2941 gggcttttat ccctcggact tcttttcagg ccccttctcg gcctttctgt gaccctgttt
3001 tttcaggccg cgaactgcgc ctggtcgggc ctgaccccca cacgctggcc ttcgtactca
3061 cgcagcctgc ccagcaggtc ggccagcgcg gcattgacca cggcggcgag gttgcggggc
3121 ggcgttccgc ctgcggtctg gtcgtgcttc acgtcgcgca gcacgcgcac cagcaccgcg
3181 cccacgtcgt cggacacgtc ccggccagcg tcggcggcgc gggtgatgtt ccaaatgagt
3241 ttccgccaga agcccagcga gtcggccccg tccccgaaag ctgccgctag cgcccgcgcc
3301 tgcctgtcca cgatctcggc gcggtaggcc gggcgggttt cccggtgggc gtcggcaagc
3361 tcccacacga cgttttcagc gagggagggc gcgggccgga cagtcatgtt atcggagggg
3421 gagggaagag cggattttat cgcccaggtg ctcacaactt cgtatgtcac ggcaaccggg
3481 gcgctttttt cgcgcagttc cccgcctgtc acatcctcac agggttcttg actctccggc
3541 acgctcttag ggagagggtg gagcatgttg tagacggtgc gcccggcctt cgcgtcggcg
3601 ttcaggtcgc gccagttccg gccccaatcg tcatgcatca ggcggacata tcccgccttc
3661 ccttccagca cgcgcctcgg cttgagcgtc acggcccaca gcgtcccggt tgccacagac
3721 tcgccgcgca ggtcgcccat gtgggcatca caggccacca gcccgacccg gcgcagatat
3781 tgaaggttct cgtagaaggc ggacttcttc agacccacat gcaccatgag cagctcggcg
3841 ggcaggtgga acacagcgcg ggtgacgtgt tccgcatagc cgcaggcgcg ggccacgtcc
3901 agcgccaccg agaacagggc gcggaagatg cgccgggccg actcgcggca gggggcgtca
3961 tctaggcagg cggtgagggt ctgcaccagt tcggcgcgtg tggcgatggt caacggcgcg
4021 ggcgcgggct gggactgctg ggcgatggtg ggcacgggcg gcggcgagat ggaccgggcc
4081 gagctggaaa tttcccgtgt gagcgcaggt ttaggcgcat ggttaccttc ctgcagtgaa
4141 atctgcgaaa tcaggcccgc ttcgaggagt cggaccatga atggattgtg tctcaagaaa
4201 aaagcctccc ctcttgtcag aggggagggt cccggtctac catgctctca gcgacgagat
4261 agctggggtt tgtgaccctg cactctgaca aagccctcac cgaaaggtgg gggtttagtc
4321 atttggccta gtaaagcaga acagaggcga tttgtcacgc aaaggcccgc cgcccgaggg
4381 gaaacaggcg gcgggccaat ggctcggcgt tgttagctgg gggtcgcacc agggaagcac
4441 agaagccctt ccctgagcag ggtttcaccg tgtggtgtcc cttcgtcctt gaaggcgaac
4501 tggtaggcca ggccgtaatg ctcctgcacc atcttttgct catccaccgc gacaaagaaa
4561 cggcgggtgt tcgccgggct gctgagggtc tgccactcgc cggacgcgcc caccgcatag
4621 gcgtctccgg ctccacattc ctcggcctgc tgatactgat acttcgtcca cagcatcacc
4681 gccttatcgg gcacctgcgc gaagctcacc agcgccccga accgcccggc gctcgtggtg
4741 aaggtggacg actgcaccat gtcggacagt tccagcgtgg caggcgcggg cgtgttcgtc
4801 ggcgcgtaca ccgtccccgc ctcgcacacg accagcgggg cggcccccgt gccgaactcg
4861 gcggcgctgc cgtttgccag cgtgagcgcc gaagacgcca ccgcgcaggt acccatgggt
4921 gccgaccacc agcaggtcca cgccgtgcgc cgcctcggtg agcacctcga ttggacggcc
4981 cctcagggtc tgaaccctcc cctgcacgcc cgcctccccg atgagtgtct gcgccgtctg
5041 ctgcgcctgc cgggcattct gctccagcat ctcccggacg tagtcgggat tcagtcccgc
5101 ccactgaacg aagccgctga ccggcggctc ctcgatcaca gtcacgacct ccagctcggc
5161 gcctgccgcc ttcccgagcg ttatggcctg cttcaaggcg cgctggctgg gctcacttcc
5221 gtcaaaggcc accacgaatc gcatccgtcc tcctccctga ccagtcaggt tcagcaagca
5281 gggtaagcct ctcccagcac tccggggctt cacacggcct tgggtctttt ctgcataccc
5341 atgagcctca gtacccgaca gtttccggca gggcttgaaa aagttcggct gacacgcgag
5401 aactggacgc gtctcccgat gtcctggcca agttgacccg ctgcttcacc gcgcacttcc
5461 cggagttccg caagaactag gtcgagctgc tctccctcat ggtcctcgcc ctccttaggg
5521 gcaaggacgt ccggcatgct aaactcgccg cgcgcttccc cggaagcgcg cacaccgcct
5581 ccgtcatccg gcgggtggaa cgcttcttcg accgtcatcc tcttcggcca gctgatgtcg
5641 cccgggtcgt tctgacgctc cttcccgccg cgcagccacg cgaatttatc cttgaccgga
5701 ccaactggaa gtatgggcag acggacgtga acgtcttgtt gctggccgtc atttggcggg
5761 acgtcgccat ccccctgctc tacgagttgc tgccccatgg gggcagcagc gacaccgaga
5821 ttcggcacac cctgatggac gatgccctgt gcctgctgtc cgccgctgac atccgggtgc
5881 tgtatgccga ccgcgaattc ccagcttgag taggacaaat ccgccgagct tcgacgagat
5941 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
6001 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
6061 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
6121 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
6181 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
6241 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
6301 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
6361 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
6421 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
6481 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
6541 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
6601 atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg
6661 gttgctacgc ctgaataagt gataataagc ggatgaatgg cagaaattcg tcgaggcggc
6721 acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg cggagaactg
6781 tgaatgcatc ccgcaagagg cccggcagt
//
