Basic Vector Information
- Vector Name:
- pMG_DV
- Length:
- 6435 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Stach C, McCann M, O'Brien C, Le TS
- Promoter:
- UbC
pMG_DV 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 ℃.
pMG_DV vector Sequence
LOCUS 62056_16780 6435 bp DNA circular SYN 23-OCT-2019
DEFINITION Cloning vector pMG_DV, complete sequence.
ACCESSION MN389527
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6435)
AUTHORS Stach C, McCann M, O'Brien C, Le TS, Somia N, Chen X, Lee K, Fu HY,
Daoutidis P, Zhao L, Hu WS, Smanski MJ.
TITLE Model-driven engineering of N-linked glycosylation in Chinese
Hamster Ovary cells
JOURNAL ACS Synth Biol (2019) In press
PUBMED 31596566
REFERENCE 2 (bases 1 to 6435)
AUTHORS Stach CS.
TITLE Direct Submission
JOURNAL Submitted (28-AUG-2019) Biochemistry, Molecular Biology, and
Biophysics; Chemical Engineering and Materials Science, University
of Minnesota, 1479 Gortner Ave, Suite 344, Saint Paul, Minnesota
55108, United States
REFERENCE 3 (bases 1 to 6435)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol (2019) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(28-AUG-2019) Biochemistry, Molecular Biology, and Biophysics;
Chemical Engineering and Materials Science, University of Minnesota,
1479 Gortner Ave, Suite 344, Saint Paul, Minnesota 55108, United
States"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..6435
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind complement(21..54)
/label=FRT (minimal)
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
misc_feature 55..65
/label=AarI site
/note="AarI site"
misc_feature 66..69
/label=Golden gate cloning scar
/note="Golden gate cloning scar"
misc_feature 70..77
/label=BbsI site
/note="BbsI site"
primer_bind 279..295
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 296..352
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(365..381)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(389..405)
/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)."
promoter complement(413..443)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(458..479)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 560..567
/label=BbsI site
/note="BbsI site"
misc_feature 568..571
/label=Golden gate cloning scar
/note="Golden gate cloning scar"
misc_feature 572..582
/label=AarI site
/note="AarI site"
promoter 588..1799
/label=UbC promoter
/note="human ubiquitin C promoter"
CDS 1831..2250
/gene="bsr"
/label=bsr
/note="Blasticidin-S deaminase from Bacillus cereus.
Accession#: P33967"
polyA_signal 2257..2305
/label=poly(A) signal
/note="synthetic polyadenylation signal"
protein_bind complement(2306..2339)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 2358..2396
/label=PhiC31 attP
/note="PhiC31 attP"
CDS complement(2719..3279)
/codon_start=1
/transl_table=11
/product="kanamycin resistance protein"
/label=kanamycin resistance protein
/protein_id="QFQ66234.1"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLHV
IYTLLLPRKYPSWLMQCGGCIRLIRLPAHSTTKRNIASSEHVLGWKPVLSIRMIWTKSI
RGSRQPNCSPGSRRACPTARISS"
terminator 3812..3898
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3990..4017
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
rep_origin 4151..4739
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(4925..5065)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(6362..6379)
/label=6xHis
/note="6xHis affinity tag"
protein_bind 6400..6433
/label=attB
/note="minimal attB site for the phi-C31 integrase (Groth
et
al., 2000)"
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