| 出品公司: | |
|---|---|
| 别名: | pET44b, pET 44b |
| 质粒类型: | 大肠杆菌蛋白表达 |
| 表达水平: | 高 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 7310 bp |
| 5' 测序引物: | T7 |
| 5' 测序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
| 3' 测序引物: | ColiDOWN |
| 3' 测序引物序列: | ColiDOWN: 5'-TTCACTTCTGAGTTCGGCATG-3' |
| 载体标签: | C-HSV, N-His, C-His, N-Thrombin, N-Nus, N-EK |
| 载体抗性: | Ampicillin |
| 表达宿主菌: | BL21(DE3) 、 Rosetta2(DE3)、C43(DE3) |
| 备注: |
Both Nterm and Cterm His tags; Nterm thrombin cleavage site; Nterm enterokinase cleavage site; a,b,c vary by MCS.
|
| 产品目录号: | ZY71123-3 |
| 稳定性: | 瞬时表达 Transient |
| 组成型: | 组成型 Constitutive |
| 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：
pET-44b(+)
- 库存：
20
- 供应商：
泽叶生物
pET44b载体基本信息
pET44b载体质粒图谱和多克隆位点信息
| Feature Name | Start | End | |
|---|---|---|---|
| T7_Terminal_primer | 69 | 87 | |
| rrnB_T2_terminator | 168 | 141 | |
| rrnB_T1_terminator | 343 | 300 | |
| rrnB_terminator | 466 | 309 | |
| EK | 694 | 680 | |
| S15 | 790 | 746 | |
| 6xHIS | 2341 | 2324 | |
| T7_transl_en_RBS | 2378 | 2362 | |
| lacO | 2423 | 2396 | |
| T7_promoter | 2441 | 2423 | |
| tet (300 - 563) | 2477 | 2740 | |
| pBRrevBam_primer | 2548 | 2529 | |
| lacI | 2823 | 3914 | |
| ROP | 4486 | 4677 | |
| pGEX_3_primer | 4693 | 4671 | |
| pBR322_origin | 5711 | 5092 | |
| AmpR_promoter | 5798 | 5826 | |
| Ampicillin | 5869 | 6729 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 2 | 2353 | 509 | |
| ORF frame 2 | 803 | 2131 | |
| ORF frame 2 | 2324 | 3082 | |
| ORF frame 3 | 2955 | 3914 | |
| ORF frame 1 | 5869 | 6729 |
| Enzyme Name | Cut |
|---|---|
| PacI | 503 |
| XhoI | 530 |
| PmlI | 576 |
| NotI | 592 |
| EagI | 592 |
| HindIII | 599 |
| KpnI | 624 |
| SalI | 626 |
| PstI | 639 |
| AscI | 641 |
| EcoRI | 654 |
| BamHI | 660 |
| SacI | 672 |
| SmaI | 720 |
| XmaI | 718 |
| SacII | 796 |
| SpeI | 827 |
| BglII | 942 |
| MscI | 1074 |
| StuI | 1127 |
| NcoI | 1402 |
| NdeI | 2352 |
| XbaI | 2390 |
| ApaI | 3393 |
| HpaI | 3688 |
| FspI | 6433 |
pET44b载体简介
The pET-44 vectors are designed for cloning and high-level expression of peptide sequences fused with the 495 aa Nus•Tag™ protein. Compared to the pET-43.1 series, the pET-44 vectors encode an additional N-terminal His•Tag. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the COLIDOWN primer (Cat. No. 70845-3). Vector encoded sequence can be completely removed when cloning into the PshA I or Sma I sites (as shown below) by cleaving the Nus•Tag fusion protein with enterokinase or thrombin, respectively.
pET44b载体序列
LOCUS pET-44 b(+) 7310 bp DNA Circular SYN
DEFINITION pET-44 b(+)
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
FEATURES Location/Qualifiers
source 1..7310
/organism="pET-44 b(+)"
/mol_type="other DNA"
misc_feature 69..87
/label="T7_Terminal_primer"
terminator complement(141..168)
/label="rrnB_T2_terminator"
terminator complement(300..343)
/label="rrnB_T1_terminator"
terminator complement(309..466)
/label="rrnB_terminator"
CDS complement(509..2353)
/label="ORF frame 2"
misc_feature complement(680..694)
/label="EK"
misc_feature complement(746..790)
/label="S15"
CDS 803..2131
/label="ORF frame 2"
CDS 2324..3082
/label="ORF frame 2"
misc_feature complement(2324..2341)
/label="6xHIS"
misc_feature complement(2362..2378)
/label="T7_transl_en_RBS"
misc_feature complement(2396..2423)
/label="lacO"
promoter complement(2423..2441)
/label="T7_promoter"
gene 2477..2740
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
misc_feature complement(2529..2548)
/label="pBRrevBam_primer"
misc_feature 2823..3914
/label="lacI"
CDS 2955..3914
/label="ORF frame 3"
misc_feature 4486..4677
/label="ROP"
misc_feature complement(4671..4693)
/label="pGEX_3_primer"
rep_origin complement(5092..5711)
/label="pBR322_origin"
promoter 5798..5826
/label="AmpR_promoter"
gene 5869..6729
/label="Ampicillin"
/gene="Ampicillin"
CDS 5869..6729
/label="ORF frame 1"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggata catatttgaa tgtatttaga aaaataaaca
121 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
181 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt
241 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
361 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
421 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
481 agcctaggaa cgcccaactt aattaacatt agtggtggtg gtggtggtgc tcgagatcct
541 ctgggtcttc aggagcgagt tctggctggc ttgcacgtgt atacagctgt gcggccgcaa
601 gcttcgaacg cgtatcgatg gtaccgtcga cgtcctgcag gcgcgcctgt acagaattcg
661 gatccacgag ctccggactc ttgtcgtcgt catcaatcgt accagaaccc gcgctgcccc
721 gggggaccag accagttggc ggtggcgagt ccatgtgctg gcgttcaaat ttcgcagcag
781 cggtttcttt acccgcggag tgatggtgat ggtgatgacc agaaccacta gtcgcttcgt
841 caccgaacca gcaaatatta cgggcagcca taatcagtgc tccggctttt tcgtcggtca
901 acccttcgat atcagccaga tcatcaatgc cctgttcggc gagatcttcc agcgtacaaa
961 cgccacgggc ggccagtttg aatgccaaat cacgatctac cccttcaagg ttcagcagat
1021 cgtcagccgg tttgttatca ccgaggcttt cttcctgggc ctgtgcaatg gtggccagtg
1081 catttttagc acgctcgcgc agtgcttcaa cggtcggctc atcaaggcct tcgatttcca
1141 acagctcttt catcggcaca taggccaatt cttccagcgt cgagaagcct tcttctacca
1201 gaacagtcgc gaagtcttcg tcgatgtcga gatatttggt gaaggtgtcg atcgctgcgt
1261 gcgcttccgc ctgatgctta gcttgcaggt cgtcaacggt catcacgttg agttcccaac
1321 cgctcagttg cgaagccaga cgcacgttct gaccgttacg gccaatcgcc tgcgccagat
1381 taccggcttc aacggcgatg tccatggtgt gtttatcttc atccaccacg atagaagcaa
1441 cgtctgccgg tgccattgcg ttaatcacga actgcgccgg gttatcatcc cacaggacga
1501 tatcgatacg ctcgccaccc agttcagtag acaccgcctg aacacgcgcg ccacgcatac
1561 ctacgcaagc acctaccgga tcgatacgtt tatcgttggt tttcaccgcg attttcgcac
1621 gagaacccgg atcgcgagcc gctgctttaa tttcaatcac ttcttcgccg atttctggca
1681 cttcaatacg gaacagttcg atcagcattt ccggcttgga acgagtgacg aacagttgcg
1741 cgccacgcgc ttccgggcga acggaataga gcacgccacg aacgcggtcg ccagggcgga
1801 agttttcacg cggcagcata tcttcgcgca ggatcacggc ttcagcgttg ttgcccagat
1861 ccagagagat gttgtcgcgg tttacttttt tcaccacgcc ggtgatgatt tcaccttcgt
1921 gttcacggaa ctgatcaacc accatcgcac gttcggcttc acgcactttc tgcacgataa
1981 cctgttttgc cgtctgggta gtgatacggt caaaggtaac agactcaatc tgatcttcaa
2041 cgtaatcgcc caggttcagg ctttcatctt cataacgtgc ggcttcaagg gtgatttcct
2101 tggtcggctg ggtgacttca tcaacaacta accagcgacg gaaagtgtca aaatcaccgc
2161 ttttgcgatc gatctgtacg cggacgtcga tctcttgttc atattttttc tttgttgctg
2221 tcgccagcgc gctttccaat gcttcgaaaa tcttctcgcg aggtagcgcc ttttcattgg
2281 atacggcttc aactacagcc aaaatttctt tgttcatact agaatggtga tggtggtgat
2341 ggctgctgcc catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga
2401 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc
2461 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
2521 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
2581 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
2641 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
2701 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
2761 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
2821 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
2881 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
2941 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
3001 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
3061 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
3121 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
3181 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
3241 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
3301 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
3361 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
3421 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
3481 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
3541 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
3601 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg
3661 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
3721 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
3781 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
3841 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
3901 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
3961 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
4021 atgactagca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
4081 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
4141 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
4201 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
4261 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
4321 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
4381 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
4441 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
4501 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
4561 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
4621 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
4681 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
4741 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
4801 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
4861 tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
4921 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
4981 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
5041 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
5101 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
5161 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
5221 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
5281 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
5341 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
5401 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
5461 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
5521 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
5581 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
5641 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
5701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
5761 tttggtcatg acctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
5821 gacaataacc ctgataaatg cttcaatcat gattgaaaaa ggaagagtat gagtattcaa
5881 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
5941 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
6001 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
6061 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
6121 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
6181 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
6241 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
6301 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
6361 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
6421 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
6481 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
6541 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
6601 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
6661 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
6721 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
6781 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgcc atcgtgccgc
6841 cagaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
6901 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
6961 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
7021 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
7081 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
7141 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
7201 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
7261 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
DEFINITION pET-44 b(+)
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
FEATURES Location/Qualifiers
source 1..7310
/organism="pET-44 b(+)"
/mol_type="other DNA"
misc_feature 69..87
/label="T7_Terminal_primer"
terminator complement(141..168)
/label="rrnB_T2_terminator"
terminator complement(300..343)
/label="rrnB_T1_terminator"
terminator complement(309..466)
/label="rrnB_terminator"
CDS complement(509..2353)
/label="ORF frame 2"
misc_feature complement(680..694)
/label="EK"
misc_feature complement(746..790)
/label="S15"
CDS 803..2131
/label="ORF frame 2"
CDS 2324..3082
/label="ORF frame 2"
misc_feature complement(2324..2341)
/label="6xHIS"
misc_feature complement(2362..2378)
/label="T7_transl_en_RBS"
misc_feature complement(2396..2423)
/label="lacO"
promoter complement(2423..2441)
/label="T7_promoter"
gene 2477..2740
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
misc_feature complement(2529..2548)
/label="pBRrevBam_primer"
misc_feature 2823..3914
/label="lacI"
CDS 2955..3914
/label="ORF frame 3"
misc_feature 4486..4677
/label="ROP"
misc_feature complement(4671..4693)
/label="pGEX_3_primer"
rep_origin complement(5092..5711)
/label="pBR322_origin"
promoter 5798..5826
/label="AmpR_promoter"
gene 5869..6729
/label="Ampicillin"
/gene="Ampicillin"
CDS 5869..6729
/label="ORF frame 1"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggata catatttgaa tgtatttaga aaaataaaca
121 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
181 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt
241 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
361 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
421 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
481 agcctaggaa cgcccaactt aattaacatt agtggtggtg gtggtggtgc tcgagatcct
541 ctgggtcttc aggagcgagt tctggctggc ttgcacgtgt atacagctgt gcggccgcaa
601 gcttcgaacg cgtatcgatg gtaccgtcga cgtcctgcag gcgcgcctgt acagaattcg
661 gatccacgag ctccggactc ttgtcgtcgt catcaatcgt accagaaccc gcgctgcccc
721 gggggaccag accagttggc ggtggcgagt ccatgtgctg gcgttcaaat ttcgcagcag
781 cggtttcttt acccgcggag tgatggtgat ggtgatgacc agaaccacta gtcgcttcgt
841 caccgaacca gcaaatatta cgggcagcca taatcagtgc tccggctttt tcgtcggtca
901 acccttcgat atcagccaga tcatcaatgc cctgttcggc gagatcttcc agcgtacaaa
961 cgccacgggc ggccagtttg aatgccaaat cacgatctac cccttcaagg ttcagcagat
1021 cgtcagccgg tttgttatca ccgaggcttt cttcctgggc ctgtgcaatg gtggccagtg
1081 catttttagc acgctcgcgc agtgcttcaa cggtcggctc atcaaggcct tcgatttcca
1141 acagctcttt catcggcaca taggccaatt cttccagcgt cgagaagcct tcttctacca
1201 gaacagtcgc gaagtcttcg tcgatgtcga gatatttggt gaaggtgtcg atcgctgcgt
1261 gcgcttccgc ctgatgctta gcttgcaggt cgtcaacggt catcacgttg agttcccaac
1321 cgctcagttg cgaagccaga cgcacgttct gaccgttacg gccaatcgcc tgcgccagat
1381 taccggcttc aacggcgatg tccatggtgt gtttatcttc atccaccacg atagaagcaa
1441 cgtctgccgg tgccattgcg ttaatcacga actgcgccgg gttatcatcc cacaggacga
1501 tatcgatacg ctcgccaccc agttcagtag acaccgcctg aacacgcgcg ccacgcatac
1561 ctacgcaagc acctaccgga tcgatacgtt tatcgttggt tttcaccgcg attttcgcac
1621 gagaacccgg atcgcgagcc gctgctttaa tttcaatcac ttcttcgccg atttctggca
1681 cttcaatacg gaacagttcg atcagcattt ccggcttgga acgagtgacg aacagttgcg
1741 cgccacgcgc ttccgggcga acggaataga gcacgccacg aacgcggtcg ccagggcgga
1801 agttttcacg cggcagcata tcttcgcgca ggatcacggc ttcagcgttg ttgcccagat
1861 ccagagagat gttgtcgcgg tttacttttt tcaccacgcc ggtgatgatt tcaccttcgt
1921 gttcacggaa ctgatcaacc accatcgcac gttcggcttc acgcactttc tgcacgataa
1981 cctgttttgc cgtctgggta gtgatacggt caaaggtaac agactcaatc tgatcttcaa
2041 cgtaatcgcc caggttcagg ctttcatctt cataacgtgc ggcttcaagg gtgatttcct
2101 tggtcggctg ggtgacttca tcaacaacta accagcgacg gaaagtgtca aaatcaccgc
2161 ttttgcgatc gatctgtacg cggacgtcga tctcttgttc atattttttc tttgttgctg
2221 tcgccagcgc gctttccaat gcttcgaaaa tcttctcgcg aggtagcgcc ttttcattgg
2281 atacggcttc aactacagcc aaaatttctt tgttcatact agaatggtga tggtggtgat
2341 ggctgctgcc catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga
2401 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc
2461 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
2521 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
2581 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
2641 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
2701 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
2761 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
2821 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
2881 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
2941 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
3001 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
3061 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
3121 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
3181 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
3241 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
3301 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
3361 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
3421 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
3481 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
3541 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
3601 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg
3661 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
3721 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
3781 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
3841 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
3901 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
3961 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
4021 atgactagca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
4081 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
4141 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
4201 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
4261 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
4321 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
4381 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
4441 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
4501 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
4561 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
4621 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
4681 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
4741 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
4801 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
4861 tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
4921 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
4981 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
5041 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
5101 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
5161 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
5221 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
5281 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
5341 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
5401 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
5461 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
5521 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
5581 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
5641 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
5701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
5761 tttggtcatg acctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
5821 gacaataacc ctgataaatg cttcaatcat gattgaaaaa ggaagagtat gagtattcaa
5881 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
5941 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
6001 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
6061 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
6121 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
6181 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
6241 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
6301 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
6361 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
6421 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
6481 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
6541 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
6601 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
6661 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
6721 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
6781 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgcc atcgtgccgc
6841 cagaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
6901 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
6961 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
7021 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
7081 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
7141 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
7201 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
7261 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
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文献和实验相关实验
表达载体的构建是生物技术和所需蛋白质产生的基本工具，本文总结了pET-32α（+）载体技术的构建，该技术通常用于研究实验室。该方法包括获得用于构建载体的外源DNA片段，将DNA片段亚克隆到pET-32α（+）表达载体中，在大肠杆菌 BL21（DE3）中进行蛋白质表达以及在大肠杆菌中在天然条件下进行蛋白质纯化。裂解液。介绍作为分子生物学实验室中非常有用的实践，重组DNA技术（或基因克隆）是指将DNA片段从一种生物体转移到自我复制的遗传元件如表达载体。然后插入的DNA可以在外源宿主细胞中繁殖
相关专题 1)噬菌体DE3溶源化的菌株如BL21(DE3)是最常用的表达菌株，噬菌体DE3是λ噬菌体的衍生株，构建好的表达载体 可以直接转入表达菌株中，诱导蛋白的表达方式与lac启动子一样都是IPTG诱导。 2)表达菌株BL21(DE3)上带有lacI基因，T7 RNA 聚合酶编码基因以及lacUV5启动子，lacUV5启动子可以启动T7 RNA 聚合酶的表达; 3)表达质粒 如pET-28质粒带有T7启动子和编码阻遏
Expression System、HisPatch ThioFusion System、pBAD Inducible Bacterial System、pL System和T7-based Systems。这四个系列的载体可谓把原核表达的发展史中出现过的几种启动子都囊括进去了，包括T7、pL、pBAD和trc启动子，同时几种经典的调控方式也都出现了，包括有乳糖操纵子、色氨酸操纵子和阿拉伯糖操纵子。 冠军系列 Champion™ pET Expression System之所以取
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pET-44b(+)载体
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