Cre-excised pCOIN DV vector (V010033#)

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

Cre-excised pCOIN DV6228 bp30060090012001500180021002400270030003300360039004200450048005100540057006000chicken beta-globin 5'HS4 insulator core sequencechicken beta-globin 5'HS4 insulator core sequencemPGK1 promotervertebrate consensus sequence for strong initiation of translation (Kozak, 1987)FRT mutantM13 revlac operatorlac promoterpMB1 oriunnamed protein product; ampicillin resistanceAmpR promoterM13 fwdFRTSASK primerloxPM13 fwdattR4unnamed protein product; ccdBunnamed protein product; chloramphenicol resistancelacUV5 promoterattR3M13 revBGH polyA

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

Copy Sequence

Download GeneBank File(.gb)

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  JOURNAL   Unpublished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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