| 货号 | 产品名称 | 规格 | |
| MF3301-2UG | pETBlue-2 Vector 大肠杆菌表达载体 | 2μg | |
| MF3301-5UG | pETBlue-2 Vector 大肠杆菌表达载体 | 5μg |
基本信息
| 载体用途： | 大肠杆菌表达载体 |
| 载体大小： | 3653bp |
| 载体标签： | C-HSV，C-6×His |
| 原核抗性： | Amp |
| 克隆菌株： | DH5α |
| 真核抗性 | LacZ |
| 启动子： | T7 |
| 复制子： | pBR322 |
| 终止子： | T7 terminator |
| 表达宿主： | BL21(DE3) |
| 载体拷贝数： | 高 |
| 5’测序引物： | T7: TAATACGACTCACTATAGGG |
| 3’测序引物： | T7-ter: TGCTAGTTATTGCTCAGCGG |
pETBlue-2 Sequence
LOCUS Exported 3653 bp ds-DNA circular SYN 17-MAR-2013
DEFINITION Vector for blue/white screening and for inducible bacterial
expression of tagged proteins.
KEYWORDS pETBlue-2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3653)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
COMMENT This vector encodes a start codon that is positioned for optimal
expression.
FEATURES Location/Qualifiers
source 1..3653
/organism=”synthetic DNA construct”
/lab_host=”Escherichia coli”
/mol_type=”other DNA”
promoter 1..19
/note=”T7 promoter”
/note=”promoter for bacteriophage T7 RNA polymerase”
protein_bind 20..44
/bound_moiety=”lac repressor encoded by lacI”
/note=”lac operator”
/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).”
CDS complement(54..491)
/codon_start=1
/gene=”lacZ (truncated)”
/product=”LacZ-alpha fragment of beta-galactosidase”
/note=”lacZ-alpha”
/translation=”MTMITTPNLINISGGGGGARDPLGLQERVLAGLHVYTAVRPQASN
AYRWYRRRPAGAPVQNSDPRAPGISPWYISFLKLNKIILLANQDLSLAVVLQRRDWENP
GVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWKL”
RBS 265..270
/note=”ribosome binding site”
CDS 278..280
/codon_start=1
/product=”start codon”
/note=”ATG”
/translation=”M”
misc_feature 283..298
/note=”blunt sites”
/note=”blunt cutter sites that allow ORF insertion in all
three reading frames”
misc_feature 377..392
/note=”blunt sites”
/note=”blunt cutter sites that allow ORF insertion in all
three reading frames”
CDS 398..430
/codon_start=1
/product=”HSV (herpes simplex virus) epitope tag”
/note=”HSV tag”
/translation=”QPELAPEDPED”
CDS 437..454
/codon_start=1
/product=”6xHis affinity tag”
/note=”6xHis”
/translation=”HHHHHH”
promoter complement(541..569)
/note=”tet promoter”
/note=”E. coli promoter for tetracycline efflux protein
gene”
terminator 739..825
/gene=”Escherichia coli rrnB”
/note=”rrnB T1 terminator”
/note=”transcription terminator T1 from the E. coli rrnB
gene”
terminator 917..944
/note=”rrnB T2 terminator”
/note=”transcription terminator T2 from the E. coli rrnB
gene”
terminator 1012..1059
/note=”T7 terminator”
/note=”transcription terminator for bacteriophage T7 RNA
polymerase”
rep_origin 1096..1551
/direction=RIGHT
/note=”f1 ori”
/note=”f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis”
CDS complement(1666..2526)
/codon_start=1
/gene=”bla”
/product=”beta-lactamase”
/note=”AmpR”
/note=”confers resistance to ampicillin, carbenicillin, and
related antibiotics”
/translation=”MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW”
rep_origin 2619..3207
/direction=RIGHT
/note=”ori”
/note=”high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication”
protein_bind 3606..3625
/bound_moiety=”lac repressor encoded by lacI”
/note=”lac operator (symmetric)”
/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). The
symmetric lac operator was optimized for tight binding of
lac repressor.”
ORIGIN
1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gacttacaat
61 ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtacg ggcctcttcg
121 ctattacgcc agcttgcgaa cggtgggtgc gctgcaaggc gattaagttg ggtaacgcca
181 ggattctccc agtcacgacg ttgtaaaacg acggccagcg agagatcttg attggctagc
241 agaataattt tgtttaactt taagaaggag atataccatg gcgatatccc gggagctcgt
301 ggatccgaat tctgtacagg cgcgcctgca ggacgtcgac ggtaccatcg atacgcgttc
361 gaagcttgcg gccgcacagc tgtatacacg tgcaagccag ccagaactcg ctcctgaaga
421 cccagaggat ctcgagcacc accaccacca ccactaatgt taattaagtt gggcgttgta
481 atcatagtca taatcaatac tcctgactgc gttagcaatt taactgtgat aaactaccgc
541 attaaagcta ttcgatgata agctgtcaaa catgataatt cttgaagacg aaagggccta
601 ggctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
661 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
721 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
781 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
841 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
901 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
961 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctgagcaata actagcataa
1021 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
1081 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
1141 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
1201 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
1261 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
1321 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
1381 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
1441 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
1501 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt tctggcggca
1561 cgatggcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
1621 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
1681 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
1741 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
1801 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
1861 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
1921 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
1981 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
2041 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
2101 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
2161 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
2221 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
2281 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
2341 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
2401 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
2461 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
2521 actcatactc ttcctttttc aatcatgacc aaaatccctt aacgtgagtt ttcgttccac
2581 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
2641 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
2701 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
2761 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
2821 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
2881 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
2941 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
3001 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
3061 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
3121 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
3181 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
3241 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
3301 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
3361 agcgagtcag tgagcgagga agccggcgat aatggcctgc ttctcgccga aacgtttggt
3421 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
3481 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
3541 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgac
3601 cggtgaattg tgagcgctca caattctcgt gacatcataa cgtcccgcga aat
保存与运输方法
保存：-20℃保存，至少1年有效。
运输：冰袋运输。
注意事项
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