| 出品公司: | |
|---|---|
| 别名: | pET44c, pET 44c |
| 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: | 7309 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: | ZY71124-3 |
| 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-44c(+)
- 库存：
20
- 供应商：
泽叶生物
pET44c载体基本信息
pET44c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 | 693 | 679 | |
| S15 | 789 | 745 | |
| 6xHIS | 2340 | 2323 | |
| T7_transl_en_RBS | 2377 | 2361 | |
| lacO | 2422 | 2395 | |
| T7_promoter | 2440 | 2422 | |
| tet (300 - 563) | 2476 | 2739 | |
| pBRrevBam_primer | 2547 | 2528 | |
| lacI | 2822 | 3913 | |
| ROP | 4485 | 4676 | |
| pGEX_3_primer | 4692 | 4670 | |
| pBR322_origin | 5710 | 5091 | |
| AmpR_promoter | 5797 | 5825 | |
| Ampicillin | 5868 | 6728 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 1 | 427 | 2130 | |
| ORF frame 2 | 2352 | 499 | |
| ORF frame 1 | 2323 | 3081 | |
| ORF frame 2 | 2954 | 3913 | |
| ORF frame 3 | 5868 | 6728 |
| 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 | 719 |
| XmaI | 717 |
| SacII | 795 |
| SpeI | 826 |
| BglII | 941 |
| MscI | 1073 |
| StuI | 1126 |
| NcoI | 1401 |
| NdeI | 2351 |
| XbaI | 2389 |
| ApaI | 3392 |
| HpaI | 3687 |
| FspI | 6432 |
pET44c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.
pET44c载体序列
LOCUS pET-44 c(+) 7309 bp DNA Circular SYN
DEFINITION pET-44 c(+)
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..7309
/organism="pET-44 c(+)"
/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 427..2130
/label="ORF frame 1"
CDS complement(499..2352)
/label="ORF frame 2"
misc_feature complement(679..693)
/label="EK"
misc_feature complement(745..789)
/label="S15"
CDS 2323..3081
/label="ORF frame 1"
misc_feature complement(2323..2340)
/label="6xHIS"
misc_feature complement(2361..2377)
/label="T7_transl_en_RBS"
misc_feature complement(2395..2422)
/label="lacO"
promoter complement(2422..2440)
/label="T7_promoter"
gene 2476..2739
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
misc_feature complement(2528..2547)
/label="pBRrevBam_primer"
misc_feature 2822..3913
/label="lacI"
CDS 2954..3913
/label="ORF frame 2"
misc_feature 4485..4676
/label="ROP"
misc_feature complement(4670..4692)
/label="pGEX_3_primer"
rep_origin complement(5091..5710)
/label="pBR322_origin"
promoter 5797..5825
/label="AmpR_promoter"
gene 5868..6728
/label="Ampicillin"
/gene="Ampicillin"
CDS 5868..6728
/label="ORF frame 3"
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 ctcggactct tgtcgtcgtc atcaatcgta ccagaacccg cgctgccccg
721 ggggaccaga ccagttggcg gtggcgagtc catgtgctgg cgttcaaatt tcgcagcagc
781 ggtttcttta cccgcggagt gatggtgatg gtgatgacca gaaccactag tcgcttcgtc
841 accgaaccag caaatattac gggcagccat aatcagtgct ccggcttttt cgtcggtcaa
901 cccttcgata tcagccagat catcaatgcc ctgttcggcg agatcttcca gcgtacaaac
961 gccacgggcg gccagtttga atgccaaatc acgatctacc ccttcaaggt tcagcagatc
1021 gtcagccggt ttgttatcac cgaggctttc ttcctgggcc tgtgcaatgg tggccagtgc
1081 atttttagca cgctcgcgca gtgcttcaac ggtcggctca tcaaggcctt cgatttccaa
1141 cagctctttc atcggcacat aggccaattc ttccagcgtc gagaagcctt cttctaccag
1201 aacagtcgcg aagtcttcgt cgatgtcgag atatttggtg aaggtgtcga tcgctgcgtg
1261 cgcttccgcc tgatgcttag cttgcaggtc gtcaacggtc atcacgttga gttcccaacc
1321 gctcagttgc gaagccagac gcacgttctg accgttacgg ccaatcgcct gcgccagatt
1381 accggcttca acggcgatgt ccatggtgtg tttatcttca tccaccacga tagaagcaac
1441 gtctgccggt gccattgcgt taatcacgaa ctgcgccggg ttatcatccc acaggacgat
1501 atcgatacgc tcgccaccca gttcagtaga caccgcctga acacgcgcgc cacgcatacc
1561 tacgcaagca cctaccggat cgatacgttt atcgttggtt ttcaccgcga ttttcgcacg
1621 agaacccgga tcgcgagccg ctgctttaat ttcaatcact tcttcgccga tttctggcac
1681 ttcaatacgg aacagttcga tcagcatttc cggcttggaa cgagtgacga acagttgcgc
1741 gccacgcgct tccgggcgaa cggaatagag cacgccacga acgcggtcgc cagggcggaa
1801 gttttcacgc ggcagcatat cttcgcgcag gatcacggct tcagcgttgt tgcccagatc
1861 cagagagatg ttgtcgcggt ttactttttt caccacgccg gtgatgattt caccttcgtg
1921 ttcacggaac tgatcaacca ccatcgcacg ttcggcttca cgcactttct gcacgataac
1981 ctgttttgcc gtctgggtag tgatacggtc aaaggtaaca gactcaatct gatcttcaac
2041 gtaatcgccc aggttcaggc tttcatcttc ataacgtgcg gcttcaaggg tgatttcctt
2101 ggtcggctgg gtgacttcat caacaactaa ccagcgacgg aaagtgtcaa aatcaccgct
2161 tttgcgatcg atctgtacgc ggacgtcgat ctcttgttca tattttttct ttgttgctgt
2221 cgccagcgcg ctttccaatg cttcgaaaat cttctcgcga ggtagcgcct tttcattgga
2281 tacggcttca actacagcca aaatttcttt gttcatacta gaatggtgat ggtggtgatg
2341 gctgctgccc atatgtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
2401 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatcg
2461 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
2521 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
2581 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
2641 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
2701 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat
2761 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
2821 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
2881 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
2941 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
3001 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
3061 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
3121 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
3181 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
3241 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
3301 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
3361 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
3421 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
3481 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
3541 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
3601 taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg acgataccga
3661 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
3721 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
3781 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
3841 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
3901 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat
3961 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
4021 tgactagcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
4081 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
4141 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
4201 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
4261 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
4321 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
4381 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
4441 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
4501 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
4561 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
4621 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
4681 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
4741 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
4801 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
4861 gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
4921 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
4981 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
5041 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
5101 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
5161 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
5221 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
5281 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
5341 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
5401 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
5461 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
5521 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
5581 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
5641 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
5701 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
5761 ttggtcatga cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
5821 acaataaccc tgataaatgc ttcaatcatg attgaaaaag gaagagtatg agtattcaac
5881 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
5941 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
6001 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
6061 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
6121 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
6181 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
6241 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
6301 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
6361 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
6421 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
6481 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
6541 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
6601 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
6661 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
6721 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
6781 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgcca tcgtgccgcc
6841 agaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
6901 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
6961 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
7021 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
7081 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
7141 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
7201 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
7261 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgcca
DEFINITION pET-44 c(+)
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..7309
/organism="pET-44 c(+)"
/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 427..2130
/label="ORF frame 1"
CDS complement(499..2352)
/label="ORF frame 2"
misc_feature complement(679..693)
/label="EK"
misc_feature complement(745..789)
/label="S15"
CDS 2323..3081
/label="ORF frame 1"
misc_feature complement(2323..2340)
/label="6xHIS"
misc_feature complement(2361..2377)
/label="T7_transl_en_RBS"
misc_feature complement(2395..2422)
/label="lacO"
promoter complement(2422..2440)
/label="T7_promoter"
gene 2476..2739
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
misc_feature complement(2528..2547)
/label="pBRrevBam_primer"
misc_feature 2822..3913
/label="lacI"
CDS 2954..3913
/label="ORF frame 2"
misc_feature 4485..4676
/label="ROP"
misc_feature complement(4670..4692)
/label="pGEX_3_primer"
rep_origin complement(5091..5710)
/label="pBR322_origin"
promoter 5797..5825
/label="AmpR_promoter"
gene 5868..6728
/label="Ampicillin"
/gene="Ampicillin"
CDS 5868..6728
/label="ORF frame 3"
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 ctcggactct tgtcgtcgtc atcaatcgta ccagaacccg cgctgccccg
721 ggggaccaga ccagttggcg gtggcgagtc catgtgctgg cgttcaaatt tcgcagcagc
781 ggtttcttta cccgcggagt gatggtgatg gtgatgacca gaaccactag tcgcttcgtc
841 accgaaccag caaatattac gggcagccat aatcagtgct ccggcttttt cgtcggtcaa
901 cccttcgata tcagccagat catcaatgcc ctgttcggcg agatcttcca gcgtacaaac
961 gccacgggcg gccagtttga atgccaaatc acgatctacc ccttcaaggt tcagcagatc
1021 gtcagccggt ttgttatcac cgaggctttc ttcctgggcc tgtgcaatgg tggccagtgc
1081 atttttagca cgctcgcgca gtgcttcaac ggtcggctca tcaaggcctt cgatttccaa
1141 cagctctttc atcggcacat aggccaattc ttccagcgtc gagaagcctt cttctaccag
1201 aacagtcgcg aagtcttcgt cgatgtcgag atatttggtg aaggtgtcga tcgctgcgtg
1261 cgcttccgcc tgatgcttag cttgcaggtc gtcaacggtc atcacgttga gttcccaacc
1321 gctcagttgc gaagccagac gcacgttctg accgttacgg ccaatcgcct gcgccagatt
1381 accggcttca acggcgatgt ccatggtgtg tttatcttca tccaccacga tagaagcaac
1441 gtctgccggt gccattgcgt taatcacgaa ctgcgccggg ttatcatccc acaggacgat
1501 atcgatacgc tcgccaccca gttcagtaga caccgcctga acacgcgcgc cacgcatacc
1561 tacgcaagca cctaccggat cgatacgttt atcgttggtt ttcaccgcga ttttcgcacg
1621 agaacccgga tcgcgagccg ctgctttaat ttcaatcact tcttcgccga tttctggcac
1681 ttcaatacgg aacagttcga tcagcatttc cggcttggaa cgagtgacga acagttgcgc
1741 gccacgcgct tccgggcgaa cggaatagag cacgccacga acgcggtcgc cagggcggaa
1801 gttttcacgc ggcagcatat cttcgcgcag gatcacggct tcagcgttgt tgcccagatc
1861 cagagagatg ttgtcgcggt ttactttttt caccacgccg gtgatgattt caccttcgtg
1921 ttcacggaac tgatcaacca ccatcgcacg ttcggcttca cgcactttct gcacgataac
1981 ctgttttgcc gtctgggtag tgatacggtc aaaggtaaca gactcaatct gatcttcaac
2041 gtaatcgccc aggttcaggc tttcatcttc ataacgtgcg gcttcaaggg tgatttcctt
2101 ggtcggctgg gtgacttcat caacaactaa ccagcgacgg aaagtgtcaa aatcaccgct
2161 tttgcgatcg atctgtacgc ggacgtcgat ctcttgttca tattttttct ttgttgctgt
2221 cgccagcgcg ctttccaatg cttcgaaaat cttctcgcga ggtagcgcct tttcattgga
2281 tacggcttca actacagcca aaatttcttt gttcatacta gaatggtgat ggtggtgatg
2341 gctgctgccc atatgtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
2401 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatcg
2461 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
2521 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
2581 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
2641 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
2701 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat
2761 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
2821 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
2881 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
2941 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
3001 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
3061 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
3121 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
3181 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
3241 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
3301 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
3361 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
3421 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
3481 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
3541 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
3601 taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg acgataccga
3661 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
3721 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
3781 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
3841 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
3901 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat
3961 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
4021 tgactagcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
4081 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
4141 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
4201 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
4261 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
4321 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
4381 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
4441 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
4501 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
4561 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
4621 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
4681 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
4741 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
4801 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
4861 gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
4921 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
4981 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
5041 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
5101 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
5161 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
5221 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
5281 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
5341 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
5401 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
5461 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
5521 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
5581 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
5641 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
5701 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
5761 ttggtcatga cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
5821 acaataaccc tgataaatgc ttcaatcatg attgaaaaag gaagagtatg agtattcaac
5881 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
5941 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
6001 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
6061 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
6121 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
6181 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
6241 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
6301 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
6361 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
6421 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
6481 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
6541 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
6601 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
6661 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
6721 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
6781 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgcca tcgtgccgcc
6841 agaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
6901 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
6961 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
7021 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
7081 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
7141 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
7201 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
7261 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgcca
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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-44c(+)载体
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