DBH-p2a-FLPo vector (V010025#)

Basic Vector Information

Basic Vector Information
Vector Name DBH-p2a-FLPo Antibiotic Resistance Ampicillin
Length 7156 bp Type Cloning vector
Source Sun JJ, Ray R.

DBH-p2a-FLPo vector Vector Map

DBH-p2a-FLPo7156 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900p15a origin5' homology armp2a linkerFLPoLox2722PGKEM7NeoRbghpALox27223' homology arm with PAM motif deletedAmpR promoterAmpR

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 ℃.

DBH-p2a-FLPo vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                7156 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector DBH-p2a-FLPo, complete sequence.
ACCESSION   KX151729
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7156)
  AUTHORS   Sun JJ, Ray R.
  TITLE     Generation of Two Noradrenergic-Specific 
            Dopamine-Beta-Hydroxylase-FLPo Knock-In Mice Using 
            CRISPR/Cas9-Mediated Targeting in Embryonic Stem Cells
  JOURNAL   PLoS ONE 11 (7), E0159474 (2016)
  PUBMED    27441631
REFERENCE   2  (bases 1 to 7156)
  AUTHORS   Sun JJ, Ray R.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-APR-2016) Neuroscience, Baylor College of Medicine, 
            One Baylor Plaza Room T707, Houston, TX 77030, USA
REFERENCE   3  (bases 1 to 7156)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7156
                     /organism="Cloning vector DBH-p2a-FLPo"
                     /mol_type="other DNA"
                     /db_xref="taxon:1883128"
     rep_origin      1..827
                     /label=p15a origin
                     /note="p15a origin"
     rep_origin      18..562
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     misc_feature    836..1835
                     /label=5' homology arm
                     /note="5' homology arm"
     misc_feature    1836..1901
                     /label=p2a linker
                     /note="p2a linker"
     CDS             1845..1901
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1902..3200
                     /codon_start=1
                     /product="FLPo"
                     /label=FLPo
                     /protein_id="ANW61891.1"
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     misc_feature    3201..3234
                     /label=Lox2722
                     /note="Lox2722"
     protein_bind    complement(3201..3234)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     regulatory      3235..3749
                     /regulatory_class="promoter"
                     /note="PGK"
     promoter        3241..3740
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     regulatory      3752..3817
                     /regulatory_class="promoter"
                     /note="EM7"
     promoter        3752..3799
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3818..4621
                     /codon_start=1
                     /product="NeoR"
                     /label=NeoR
                     /protein_id="ANW61892.1"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      4635..4937
                     /regulatory_class="other"
                     /note="bghpA"
     polyA_signal    4659..4883
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    4938..4971
                     /label=Lox2722
                     /note="Lox2722"
     protein_bind    complement(4938..4971)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     misc_feature    4972..5968
                     /label=3' homology arm with PAM motif deleted
                     /note="3' homology arm with PAM motif deleted"
     promoter        6044..6148
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6149..7009
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ANW61890.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"

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