Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | Cre-excised pCOIN DV | Antibiotic Resistance | Ampicillin |
| Length | 6228 bp | Type | Mammalian vector |
| Source | De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R. | ||
Cre-excised pCOIN 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 ℃.
Cre-excised pCOIN DV vector Sequence
LOCUS Exported 6228 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Mammalian vector Cre-excised pCOIN DV, complete sequence.
ACCESSION LT726873
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6228)
AUTHORS De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman
Fonseca M, Vanhoucke M, Beyaert R.
TITLE BCCM/LMBP Plasmid collection
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6228)
AUTHORS De Schamphelaire W.
TITLE Direct Submission
JOURNAL Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent,
Technologiepark 927, 9052, BELGIUM
REFERENCE 3 (bases 1 to 6228)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6228
/organism="Mammalian vector Cre-excised pCOIN DV"
/mol_type="other DNA"
/note="BCCM/LMBP Plasmid collection (Ghent
University,Belgium) accession number LMBP 8191."
/db_xref="taxon:1945163"
misc_feature 35..277
/label=chicken beta-globin 5'HS4 insulator core sequence
/note="chicken beta-globin 5'HS4 insulator core sequence"
misc_feature 278..520
/label=chicken beta-globin 5'HS4 insulator core sequence
/note="chicken beta-globin 5'HS4 insulator core sequence"
regulatory 539..1046
/regulatory_class="promoter"
/note="mPGK1 promoter"
promoter 546..1045
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
regulatory 1059..1068
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
misc_feature 1070..1117
/label=FRT mutant
/note="FRT mutant"
primer_bind complement(1162..1178)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1186..1202
/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)."
regulatory complement(1205..1288)
/regulatory_class="promoter"
/note="lac promoter"
promoter complement(1210..1240)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1255..1276
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1380..1782)
/direction=LEFT
/label=pMB1 ori
/note="pMB1 ori"
rep_origin complement(1564..2152)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2323..3183)
/codon_start=1
/note="unnamed protein product; ampicillin resistance"
/protein_id="SJL86737.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3184..3288)
/gene="bla"
/label=AmpR promoter
primer_bind 3762..3778
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 3801..3848
/label=FRT
/note="FRT"
protein_bind 3801..3848
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
misc_feature 3865..3968
/label=SA
/note="SA"
primer_bind 4001..4017
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(4031..4064)
/label=loxP
/note="loxP"
protein_bind complement(4031..4064)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind 4139..4155
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory complement(4162..4286)
/regulatory_class="attenuator"
/note="attR4"
protein_bind 4162..4286
/gene="mutant version of attR"
/label=attR4
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(4327..4632)
/codon_start=1
/note="unnamed protein product; ccdB"
/protein_id="SJL86738.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(4974..5633)
/codon_start=1
/note="unnamed protein product; chloramphenicol resistance"
/protein_id="SJL86739.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
regulatory complement(5682..5735)
/regulatory_class="promoter"
/note="lacUV5 promoter"
promoter complement(5687..5717)
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
regulatory 5743..5867
/regulatory_class="attenuator"
/note="attR3"
protein_bind complement(5743..5866)
/gene="mutant version of attR"
/label=attR3
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(5894..5910)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
regulatory 5959..6183
/regulatory_class="terminator"
/note="BGH polyA"
polyA_signal 5959..6183
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
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