pET-17b
Search name
pET-17b,Plasmid pET-17b,pET-17b vector
pET-17b Information
Promoter: T7
Replicon: ColE1 ori
Terminator: T7 Terminator
Plasmid classification: large intestine plasmid; large intestine expression plasmid; pET series plasmid.
Plasmid size: 3306bp
Plasmid tagging: N-T7
Prokaryotic resistance: ampicillin Amp (100 g/ml)
Cloning strains: E. coli DH5 and E.
Culture conditions: 37 C, aerobic, LB
Expression host: E. coli BL21 (DE3).
Culture conditions: 37 C, aerobic, LB
Induction: IPTG or lactose and its analogues.
5'sequencing primers: T7 (TAATACGACTCACTATAGGG)
3'sequencing primers: T7-ter (TGCTAGTTATTGCTCAGCGG)
pET-17b Description
PET-17b plasmid is a prokaryotic expression vector, and the N end contains a T7 tag. The plasmid contains several commonly used restriction sites to facilitate cloning of different genes. The expression was induced by T7 RNA polymerase provided by host cells. The target gene was cloned into plasmid vector and controlled by strong transcription and translation signals of bacteriophages.
PET system is the most powerful system to express recombinant proteins in E. coli, and it is also the most widely used system in prokaryotic expression nowadays. The plasmids can easily reduce protein expression by decreasing the concentration of inducers. Under non inducible conditions, the target gene can be completely silenced without transcribing.
The pET-17b vector carries an N-terminal 11aa T7?Tag? sequence followed by a region of useful cloning sites. Included in the multiple cloning region are dual BstX I sites, which allow efficient cloning using an asymmetric linker. Unique sites (except for the two BstX I 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 circluar map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below.
pET-17b Multiple cloning site
pET-17b Sequence
LOCUS Exported 3306 bp ds-DNA circular SYN 15-JUN-2017
DEFINITION synthetic circular DNA.
KEYWORDS pET-17b
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3306)
TITLE Direct Submission
JOURNAL Exported Thursday, June 15, 2017
FEATURES Location/Qualifiers
source 1..3306
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 104..208
/gene="bla"
/note="AmpR promoter"
CDS 209..1069
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1240..1828
/direction=RIGHT
/note="ori"
/note="high-copy-number colE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2258..2449)
/codon_start=1
/gene="rop"
/product="Rop protein"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
promoter 2958..2976
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 3038..3070
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/note="T7 tag (gene 10 leader)"
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
misc_feature 3079..3166
/note="mcs"
terminator 3232..3279
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
2941 tctcgatccc gcgaaattaa tacgactca