LOCUS       Exported                5894 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTurbo-Cre, complete sequence.
ACCESSION   AF334827
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5894)
  AUTHORS   Lu ZH, Graubert TA, Ley TJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JAN-2001) Division of Oncology, Section of Stem Cell 
            Biology, Washington University School of Medicine, 660 South Euclid 
            Avenue, Campus Box 8007, St. Louis, MO 63110-1093, USA
REFERENCE   2  (bases 1 to 5894)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5894
                     /organism="Cloning vector pTurbo-Cre"
                     /mol_type="other DNA"
                     /db_xref="taxon:152298"
     regulatory      1..364
                     /regulatory_class="enhancer"
                     /note="cytomegalovirus immediate early region"
     regulatory      365..646
                     /regulatory_class="promoter"
                     /note="chicken beta-actin"
     promoter        368..645
                     /label=chicken beta-actin promoter
     intron          647..1664
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     intron          647..1619
                     /note="chicken beta-actin"
     intron          1620..1664
                     /note="rabbit beta-globin"
     CDS             1742..2797
                     /codon_start=1
                     /product="cre recombinase"
                     /label=cre recombinase
                     /note="contains nuclear localization signal"
                     /protein_id="AAK11472.1"
                     /translation="MAPKKKRKVSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAF
                     SEHTWKMLLSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLN
                     MLHRRSGLPRPSDSNAVSLVVRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRC
                     QDIRNLAFLGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALS
                     LGVTKLVERWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIY
                     GAKDDSGQRYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSET
                     GAMVRLLQDGD"
     misc_feature    1742..1768
                     /note="Region: nuclear localization signal; NLS"
     CDS             1748..1768
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1769..2797
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVVRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLQD
                     GD"
     regulatory      2818..3344
                     /regulatory_class="polyA_signal_sequence"
                     /note="rabbit beta-globin"
     polyA_site      2963..3018
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(3379..3395)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3403..3419
                     /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(3427..3457)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3472..3493
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3552..3747
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3598..3733
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3753..3887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4126..4714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4885..5745)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="AAK11473.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5746..5850)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5881..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"