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"