pET22-EGFP大肠杆菌绿色荧光分泌型表达载体pET22b-EGFP-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心
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- 货 号:pET22-EGFP大肠杆菌绿色荧光分泌型表达载体pET22b-EGFP
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pET22-EGFP大肠杆菌绿色荧光分泌型表达载体
Alt Name: pET22b-EGFP plasmid
BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心www.biovector.net
启动子: | T7/lac |
---|---|
复制子: | ColE1 ori,F1 ori |
终止子: | T7 terminator |
质粒分类: | 大肠杆菌载体;PET系列表达质粒 |
质粒大小: | 6232bp |
质粒标签: | N-pelB,EGFP |
原核抗性: | 氨苄青霉素Amp |
克隆菌株: | DH5α |
培养条件: | 37℃,有氧,LB |
表达宿主: | BL21(DE3) |
诱导方式: | IPTG或乳糖及其类似物 |
5'测序引物: | T7:TAATACGACTCACTATAGGG |
3'测序引物: | T7-ter:TGCTAGTTATTGCTCAGCGG |
备注: | 分泌表达 |
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pET22b-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6232)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4
http://www.biovector.net
FEATURES Location/Qualifiers
source 1..6232
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 12..467
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 494..598
/gene="bla"
/note="AmpR promoter"
CDS 599..1459
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1630..2218
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2404..2546
/note="bom"
/note="basis of mobility region from pBR322"
CDS complement(2648..2839)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3648..4730)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(4731..4808)
/gene="lacI"
/note="lacI promoter"
promoter 5117..5135
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5136..5160
/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)."
RBS 5175..5197
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5205..5270
/codon_start=1
/gene="pelB (fragment)"
/product="leader peptide for secretion"
/note="pelB signal sequence"
/translation="MKYLLPTAAAGLLLLAAQPAMA"
CDS 5303..6022
/codon_start=1
/product="enhanced GFP"
/note="EGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 6076..6093
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 6160..6207
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
3301 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
3361 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
3421 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
3481 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
3541 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
3601 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
3661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
3721 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
3781 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
3841 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
3901 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
3961 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
4021 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
4081 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
4141 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
4201 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
4261 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
4321 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
4381 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
4441 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
4501 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
4561 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
4621 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
4681 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
4741 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
4801 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
4861 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
4921 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
4981 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
5041 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
5101 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
5161 cctctagaaa taattttgtt taactttaag aaggagatat acatatgaaa tacctgctgc
5221 cgaccgctgc tgctggtctg ctgctcctcg ctgcccagcc ggcgatggcc atggatatcg
5281 gaattaattc ggatccgaat tcatggtgag caagggcgag gagctgttca ccggggtggt
5341 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
5401 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
5461 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
5521 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
5581 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
5641 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
5701 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
5761 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
5821 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
5881 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
5941 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
6001 catggacgag ctgtacaagt aagcggccgc tcgagtctag agggccctat tctaaagctt
6061 gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
6121 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
6181 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg at
//
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