Basic Vector Information
- Vector Name:
- pENTR-BsaI-Tet-LI
- Antibiotic Resistance:
- Tetracycline
- Length:
- 4308 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Chiasson D, Gimenez-Oya V, Bircheneder M, Bachmaier S
pENTR-BsaI-Tet-LI vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pENTR-BsaI-Tet-LI vector Sequence
LOCUS 62056_9455 4308 bp DNA circular SYN 29-JUL-2019
DEFINITION Cloning vector pENTR-BsaI-Tet LI, complete sequence.
ACCESSION MK495753
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4308)
AUTHORS Chiasson D, Gimenez-Oya V, Bircheneder M, Bachmaier S, Studtrucker
T, Ryan J, Sollweck K, Leonhardt H, Boshart M, Dietrich P, Parniske
M.
TITLE A unified multi-kingdom Golden Gate cloning platform
JOURNAL Sci Rep 9 (1), 10131 (2019)
PUBMED 31300661
REFERENCE 2 (bases 1 to 4308)
AUTHORS Chiasson D.
TITLE Direct Submission
JOURNAL Submitted (06-FEB-2019) Biology, LMU Munich, Grosshaderner Str. 2-4,
Martinsried 82152, Germany
REFERENCE 3 (bases 1 to 4308)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep";
date: "2019"; volume: "9"; issue: "1"; pages: "10131"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-FEB-2019) Biology, LMU Munich, Grosshaderner Str. 2-4,
Martinsried 82152, Germany"
FEATURES Location/Qualifiers
source 1..4308
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 81..1268
/codon_start=1
/label=TcR
/note="tetracycline efflux protein"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
rep_origin 1305..1893
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator complement(2223..2250)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(2342..2428)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 2492..2508
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2524..2623
/label=attL1
/note="recombination site for the Gateway(R) LR reaction"
promoter 2687..2717
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 2771..3427
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 3772..4074
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(4142..4241)
/label=attL2
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(4259..4277)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
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