Basic Vector Information
- Vector Name:
- sh-AFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7716 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Han X, Yang J, Zeng F, Weng J
- Promoter:
- hPGK
sh-AFP 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 ℃.
sh-AFP vector Sequence
LOCUS 62056_23305 7716 bp DNA circular SYN 26-MAY-2020
DEFINITION Cloning vector sh-AFP, complete sequence.
ACCESSION MN996871
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7716)
AUTHORS Han X, Yang J, Zeng F, Weng J, Zhang Y, Peng Q, Shen L, Ding S, Liu
K, Gao Y.
TITLE Programmable Synthetic Protein Circuits for the Identification and
Suppression of Hepatocellular Carcinoma
JOURNAL Mol Ther Oncolytics 17, 70-82 (2020)
PUBMED 32322664
REFERENCE 2 (bases 1 to 7716)
AUTHORS Liu K, Yang J, Ding S.
TITLE Direct Submission
JOURNAL Submitted (27-JAN-2020) Second Department of Hepatobiliary Surgery,
Zhujiang Hospital, No. 253, Industrial Avenue, Haizhu District,
Guangzhou, Guangdong 510245, China
REFERENCE 3 (bases 1 to 7716)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther
Oncolytics 17, 70-82 (2020)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(27-JAN-2020) Second Department of Hepatobiliary Surgery, Zhujiang
Hospital, No. 253, Industrial Avenue, Haizhu District, Guangzhou,
Guangdong 510245, China"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7716
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1..792
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
misc_feature 814..1402
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 1474..1707
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 1785..1919
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 1946..2081
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(2102..2120)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2130..2146)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 2288..2743
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2769..2873
/label=AmpR promoter
CDS 2874..3731
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3905..4493
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 4781..4802
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4817..4847
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4855..4871
/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)."
primer_bind 4879..4895
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 4916..4934
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 4962..5188
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
misc_feature 5416..5541
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6034..6267
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 6452..6496
/codon_start=1
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
/translation="KNEQELLELDKWASL"
promoter 6674..6914
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_feature 7019..7136
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 7191..7695
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
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