LOCUS       Exported                9675 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pnlslacZ-ACN, complete sequence.
ACCESSION   EU530629
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9675)
  AUTHORS   Wu S, Ying G, Wu Q, Capecchi MR.
  TITLE     A protocol for constructing gene targeting vectors: generating 
            knockout mice for the cadherin family and beyond
  JOURNAL   Nat Protoc 3 (6), 1056-1076 (2008)
  PUBMED    18546598
REFERENCE   2  (bases 1 to 9675)
  AUTHORS   Wu S, Capecchi MR.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-FEB-2008) Howard Hughes Medical Institute, University 
            of Utah, 15N 2030E, Salt Lake City, UT 84112, USA
REFERENCE   3  (bases 1 to 9675)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9675
                     /organism="Cloning vector pnlslacZ-ACN"
                     /mol_type="other DNA"
                     /db_xref="taxon:512284"
     promoter        34..52
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             84..3170
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /protein_id="ACC91783.1"
                     /translation="MDPVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQ
                     LRSLNGEWRFAWFPAPEAVPESWLECDLPEAVPKKKRKVEADTVVVPSNWQMHGYDAPI
                     YTNVTYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRW
                     VGYGQDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHK
                     PTTQISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGG
                     EIIDERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREV
                     RIENGLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPN
                     HPLWYTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSV
                     IIWSLGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPF
                     PAVPKWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWV
                     DQSLIKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFR
                     LSGQTIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPE
                     SAGQLWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFC
                     IELGNKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWV
                     ERWKAAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAI
                     TVDVEVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDM
                     YTPYVFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWL
                     NIDGFHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     gene            84..3170
                     /gene="lacZ"
                     /label=lacZ
     CDS             315..335
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_site      3184..3578
                     /label=-beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     misc_recomb     3590..3623
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(3590..3623)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     polyA_signal    3650..3698
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      complement(3650..3697)
                     /regulatory_class="polyA_signal_sequence"
                     /note="HSV thymidine kinase polyA signal"
     CDS             complement(3705..4508)
                     /codon_start=1
                     /gene="neo"
                     /product="neomycin resistance protein"
                     /label=neo
                     /protein_id="ACC91784.1"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(3705..4508)
                     /gene="neo"
                     /label=neo
     regulatory      complement(4525..5231)
                     /regulatory_class="promoter"
                     /note="RNA polymerase II large subunit promoter"
     polyA_signal    5247..5295
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      5247..5294
                     /regulatory_class="polyA_signal_sequence"
                     /note="HSV thymidine kinase polyA signal"
     CDS             complement(join(5441..5644,5711..6559))
                     /codon_start=1
                     /gene="cre"
                     /product="Cre recombinase"
                     /label=cre
                     /protein_id="ACC91785.1"
                     /translation="MPKKKRKVSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFS
                     EHTWKMLLSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNM
                     LHRRSGLPRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQ
                     DIRNLAFLGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSL
                     GVTKLVERWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYG
                     AKDDSGQRYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETG
                     AMVRLLEDGD"
     gene            complement(5441..6559)
                     /gene="cre"
                     /label=cre
     intron          complement(5645..5710)
                     /note="SV40 T-antigen intron"
     CDS             complement(6536..6556)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      complement(6560..7258)
                     /regulatory_class="promoter"
                     /note="testis-specific angiotensin-converting enzyme 
                     promoter (tACE)"
     misc_recomb     7266..7299
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(7266..7299)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(7363..7381)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        7727..7831
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7832..8692
                     /codon_start=1
                     /gene="bla"
                     /product="ampicillin resistance protein"
                     /label=bla
                     /protein_id="ACC91782.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            7832..8692
                     /gene="bla"
                     /label=bla
     rep_origin      8863..9451
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"