Basic Vector Information
FU-tet-o-hOct4 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 ℃.
FU-tet-o-hOct4 vector Sequence
LOCUS Exported 9471 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS FU-tet-o-hOct4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9471)
AUTHORS Maherali N, Ahfeldt T, Rigamonti A, Utikal J, Cowan C, Hochedlinger
K
TITLE A high-efficiency system for the generation and study of human
induced pluripotent stem cells.
JOURNAL Cell Stem Cell. 2008 Sep 11. 3(3):340-5.
PUBMED 18786420
REFERENCE 2 (bases 1 to 9471)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Stem
Cell. 2008 Sep 11. 3(3):340-5."
FEATURES Location/Qualifiers
source 1..9471
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(47..66)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 238..617
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 619..817
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 772..792
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 835..1015
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1680..1913
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2098..2142
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2440..2557
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 2610..2880
/label=tetracycline response element
/bound_moiety="tetracycline repressor TetR"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 2913..2950
/label=minimal CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2947..2971
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 3050..3059
/regulatory_class="other"
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
misc_feature 4163..4751
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4216..4236)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4634..4645)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(4754..4770)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4755..4771)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 5276..5456
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(5482..5499)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5488..5712
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 5758..6186
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5845..5864)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6055..6076
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6195..6215)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 6200..6529
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6380..6515
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6442..6461
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 6577..6624
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 6643..7017
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 7147..7268
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7184..7203)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7238..7257
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(7317..7333)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7317..7333)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7330..7352)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7341..7357
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(7365..7395)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7410..7431
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7548..7565)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7719..8307)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7799..7818)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8478..9338)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 9101..9120
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9339..9443)
/gene="bla"
/label=AmpR promoter
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