LOCUS       Exported               11491 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTX-NLS-Cre, complete sequence.
ACCESSION   JQ033384
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11491)
  AUTHORS   Linkner J, Nordholz B, Junemann A, Winterhoff M, Faix J.
  TITLE     Highly effective removal of floxed Blasticidin S resistance 
            cassettes from Dictyostelium discoideum mutants by extrachromosomal 
            expression of Cre
  JOURNAL   Eur. J. Cell Biol. (2011) In press
  PUBMED    22154549
REFERENCE   2  (bases 1 to 11491)
  AUTHORS   Linkner J, Faix J.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-NOV-2011) Institute for Biophysical Chemistry, 
            Hannover Medical School, Carl-Neuberg-Str. 1, Hannover 30625, 
            Germany
REFERENCE   3  (bases 1 to 11491)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11491
                     /organism="Cloning vector pTX-NLS-Cre"
                     /mol_type="other DNA"
                     /db_xref="taxon:1127730"
     promoter        complement(12..30)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(51..67)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    75..91
                     /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)."
     promoter        complement(99..129)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    144..165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(453..1041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1212..2072)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AEX09391.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2073..2177)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2203..2658)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2800..2816
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2826..2844
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3194..3217
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             3230..4282
                     /codon_start=1
                     /gene="cre"
                     /product="NLS-Cre"
                     /label=cre
                     /note="Cre recombinase fused to nuclear localization 
                     sequence"
                     /protein_id="AEX09390.1"
                     /translation="MPKKKRKVSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFS
                     EHTWKMLLSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNM
                     LHRRSGLPRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQ
                     DIRNLAFLGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSL
                     GVTKLVERWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYG
                     AKDDSGQRYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETG
                     AMVRLLQDGD"
     gene            3230..4282
                     /gene="cre"
                     /label=cre
     CDS             3233..3253
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5078..5908
                     /codon_start=1
                     /product="NeoR"
                     /label=NeoR
                     /protein_id="AEX09392.1"
                     /translation="MDGEDVQAGSFRMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSD
                     AAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWL
                     LLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAG
                     LVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCG
                     RLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5114..5908
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"