Basic Vector Information
EF-PDGF 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 ℃.
EF-PDGF vector Sequence
LOCUS 40924_715 11831 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector EF-PDGF, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11831)
AUTHORS Linger RJ, Belikoff EJ, Yan Y, Li F, Wantuch HA, Fitzsimons HL,
Scott MJ.
TITLE Towards next generation maggot debridement therapy: transgenic
Lucilia sericata larvae that produce and secrete a human growth
factor
JOURNAL BMC Biotechnol. 16 (1), 30 (2016)
PUBMED 27006073
REFERENCE 2 (bases 1 to 11831)
AUTHORS Linger RJ, Belikoff EJ, Yan Y, Li F, Wantuch HA, Fitzsimons HL,
Scott MJ.
TITLE Direct Submission
JOURNAL Submitted (10-OCT-2015) Entomology, North Carolina State University,
112 Derieux Place, Raleigh, NC 27695-7614, USA
REFERENCE 3 (bases 1 to 11831)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11831)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BMC
Biotechnol."; date: "2016"; volume: "16"; issue: "1"; pages: "30"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(10-OCT-2015) Entomology, North Carolina State University, 112
Derieux Place, Raleigh, NC 27695-7614, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..11831
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(558..578)
/codon_start=1
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
intron complement(708..773)
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
protein_bind 1555..1833
/label=tetracycline response element
/note="contains seven copies of the tetracycline operator
tetO"
CDS 4458..5132
/codon_start=1
/label=DsRed-Express2
/note="noncytotoxic tetrameric variant of DsRed fluorescent
protein (Strack et al., 2008)"
/translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
LFQ"
protein_bind 6627..6726
/label=phage phi-C31 attP
/note="attachment site of phage phi-C31"
promoter complement(7767..7797)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(7812..7833)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(8121..8709)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8883..9740)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(9741..9845)
/label=AmpR promoter
primer_bind 10319..10335
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
polyA_signal complement(join(11830..11831,1..133))
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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