Basic Vector Information
- Vector Name:
- pMJc01
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5535 bp
- Type:
- Cloning vector
- Replication origin:
- RSF1010 oriV
- Source/Author:
- Jutersek M, Dolinar M.
- Promoter:
- lac UV5
pMJc01 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 ℃.
pMJc01 vector Sequence
LOCUS 62056_17005 5535 bp DNA circular SYN 22-JAN-2021
DEFINITION Cloning vector pMJc01, complete sequence.
ACCESSION MN201591
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5535)
AUTHORS Jutersek M, Dolinar M.
TITLE A new vector for cyanobacterial biotechnology assayed with cystatin,
a distinct non-fluorescent reporter protein
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5535)
AUTHORS Jutersek M, Dolinar M.
TITLE Direct Submission
JOURNAL Submitted (20-JUL-2019) Faculty of Chemistry and Chemical
Technology, Vecna pot 113, Ljubljana SI-1000, Slovenia
REFERENCE 3 (bases 1 to 5535)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(20-JUL-2019) Faculty of Chemistry and Chemical Technology, Vecna
pot 113, Ljubljana SI-1000, Slovenia"
FEATURES Location/Qualifiers
source 1..5535
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(9..403)
/direction=LEFT
/label=RSF1010 oriV
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
protein_bind 473..494
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 509..539
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
protein_bind 547..563
/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)."
CDS 599..1567
/label=RSF1010 RepB
/note="replication protein B of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS 1631..1843
/codon_start=1
/transl_table=11
/product="hypothetical protein"
/label=hypothetical protein
/note="OrfE"
/protein_id="QGN03459.1"
/translation="MEYEKSASGSVYLIKSDKGYWLPGGFGYTSNKAEAGRFSVADMAS
LNLDGCTLSLFREDKPFGPGKFLGD"
CDS 1845..2051
/codon_start=1
/transl_table=11
/product="hypothetical protein"
/label=hypothetical protein
/note="OrfF"
/protein_id="QGN03460.1"
/translation="MKDQKDKQTGDLLASPDAVRQARYAERMKAKGMRQRKFWLTDDEY
EALRECLEELRAAQGGGSDPASA"
CDS 2081..2917
/label=RSF1010 RepA
/note="replication protein A of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS 2907..3755
/label=RSF1010 RepC
/note="replication protein C of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS 3879..4691
/label=KanR
/note="aminoglycoside phosphotransferase"
terminator complement(4852..4910)
/label=his operon terminator
/note="This putative transcriptin terminator from the E.
coli his operon has a 2-bp deletion introduced during
synthesis. Its efficiency has not been determined."
misc_feature complement(4911..4931)
/label=BioBrick suffix
/note="universal suffix for all parts"
CDS complement(4935..4952)
/label=6xHis
/note="6xHis affinity tag"
regulatory complement(5310..5319)
/regulatory_class="ribosome_binding_site"
protein_bind complement(5363..5381)
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature complement(5388..5409)
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
'ATG'"
terminator 5412..5455
/label=bacterial terminator
/note="putative bacterial transcription terminator"
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