Basic Vector Information
L149 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 ℃.
L149 vector Sequence
LOCUS 40924_1529 9502 bp DNA circular SYN 17-DEC-2018
DEFINITION Retroviral expression vector L149, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9502)
AUTHORS Micklem DR, Lorens JB.
TITLE Direct Submission
JOURNAL Submitted (27-MAY-2008) Department of Biomedicine, University of
Bergen, Jonas Lies vei 91, Bergen 5009, Norway
REFERENCE 2 (bases 1 to 9502)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 9502)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
(27-MAY-2008) Department of Biomedicine, University of Bergen, Jonas
Lies vei 91, Bergen 5009, Norway"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9502
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 62..365
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 366..569
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 601..609
/label=R region 5'LTR
/note="R region 5'LTR"
misc_feature 670..745
/label=U5 region 5'LTR
/note="U5 region 5'LTR"
misc_feature 808..1165
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1230..1646
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
misc_feature 1656..2030
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
polyA_signal complement(2049..2256)
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
misc_feature complement(2946..3029)
/gene="P2AG2ALuc2ANTR"
/label=FMDV 2A self-cleaving linker region
/note="FMDV 2A self-cleaving linker region"
CDS complement(3030..4679)
/label=luciferase
/note="firefly luciferase"
misc_feature complement(4680..4763)
/gene="P2AG2ALuc2ANTR"
/label=FMDV 2A self-cleaving linker region
/note="FMDV 2A self-cleaving linker region"
CDS complement(4764..5480)
/label=EGFP
/note="enhanced GFP"
misc_feature 5481..5549
/label=FMDV 2A self-cleaving linker region
/note="FMDV 2A self-cleaving linker region"
CDS complement(5553..6146)
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPG"
intron complement(6230..6362)
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
regulatory complement(6465..6517)
/label=PhCMV* minimal promoter
/note="PhCMV* minimal promoter"
/regulatory_class="promoter"
promoter complement(6480..6517)
/label=minimal CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind complement(6550..6820)
/label=tetracycline response element
/note="contains seven copies of the tetracycline operator
tetO"
misc_feature 6892..7056
/label=U3SIN (self-inactivating) region
/note="U3SIN (self-inactivating) region"
repeat_region 6892..7056
/label=MoMLV 3'LTR, self-inactivating
/note="MoMLV 3'LTR, self-inactivating"
misc_feature 7058..7125
/label=R region 3'LTR
/note="R region 3'LTR"
primer_bind complement(7278..7294)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(7302..7318)
/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(7326..7356)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(7371..7392)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(7680..8268)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8445..9302)
/label=AmpR
/note="beta-lactamase"
promoter complement(9303..9407)
/label=AmpR promoter
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