The pET-12a-c vectors carry an N-terminal ompT sequence for potential periplasmic export of target proteins. 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 circular map. The cloning/ expression region of the coding strand transcribed by T7 RNA polymerase is shown below.
| 载体基本信息 | |||
| 载体名称 | pET-12a | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 4674 bp | 载体类型 | E. coli Expression Vectors |
| 拷贝数 | High copy number | 克隆方法 | Enzyme digestion and ligation |
| 5引物 | 5-TAATACGACTCACTATAGGG-3 | ||
pET-12a 载体载体图谱
pET-12a 载体载体序列
LOCUS Exported 4674 bp ds-DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4674)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4674
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 10..38
/note="tet promoter"
/note="E. coli promoter for tetracycline efflux protein
gene"
terminator 404..451
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
RBS 589..611
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
promoter complement(643..661)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 2226..2417
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 2519..2661
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(2847..3435)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3606..4466)
/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"
promoter complement(4467..4571)
/gene="bla"
/note="AmpR promoter"
