Basic Vector Information
- Vector Name:
- pNIL2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4504 bp
- Type:
- Bacterial expression vector
- Replication origin:
- ori
- Source/Author:
- De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R.
pNIL2 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 ℃.
pNIL2 vector Sequence
LOCUS 40924_33277 4504 bp DNA circular SYN 18-DEC-2018
DEFINITION Bacterial expression vector pNIL2, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4504)
AUTHORS De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman
Fonseca M, Vanhoucke M, Beyaert R.
TITLE BCCM/LMBP Plasmid collection
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4504)
AUTHORS De Schamphelaire W.
TITLE Direct Submission
JOURNAL Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent,
Technologiepark 927, 9052, BELGIUM
REFERENCE 3 (bases 1 to 4504)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4504)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927,
9052, BELGIUM"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4504
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 27..75
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
regulatory 134..215
/label=fd terminator
/note="fd terminator"
/regulatory_class="terminator"
terminator 137..185
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
rep_origin 313..768
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin complement(927..1515)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1689..2546)
/label=AmpR
/note="beta-lactamase"
misc_feature complement(2861..3520)
/label=chloramphenicol sensitive
/note="chloramphenicol sensitive"
CDS complement(2861..3520)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTV*LDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(3521..3623)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
promoter 3788..3816
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 3824..3840
/label=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)."
misc_feature 3865..3881
/label=5' UTR of ompA, incl. RBS
/note="5' UTR of ompA, incl. RBS"
sig_peptide 3882..3944
/label=OmpA signal peptide
/note="signal peptide from the E. coli outer membrane
protein OmpA"
CDS 3945..4394
/codon_start=1
/note="unnamed protein product; mIL2 mature sequence"
/protein_id="SJL88449.1"
/translation="APTSSSTSSSTAEAQQQQQQQQQQQQHLEQLLMDLQELLSRMENY
RNLKLPRMLTFKFYLPKQATELKDLQCLEDELGPLRHVLDLTQSKSFQLEDAENFISNI
RVTVVKLKGSDNTFECQFDDESATVVDFLRRWIAFCQSIISTSPQ"
misc_feature 4395..4497
/label=3' UTR of mIL2
/note="3' UTR of mIL2"
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