LOCUS       Exported                5214 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cre expression vector pCS2-olCre DNA, complete sequence.
ACCESSION   LC094445
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5214)
  AUTHORS   Kishimoto K.
  TITLE     Two useful site-specific recombination system VCre/VloxP and 
            SCre/SloxP in Medaka
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5214)
  AUTHORS   Kishimoto K, Kinoshita M.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-NOV-2015) Contact:Kenta Kishimoto Kyoto University, 
            Graduate School of Agriculture; Kitashirakawa Oiwaketyo, Sakyo, 
            Kyoto 606-8502, Japan URL :http://www.kais.kyoto-u.ac.jp/english/
REFERENCE   3  (bases 1 to 5214)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5214
                     /organism="Cre expression vector pCS2-olCre"
                     /mol_type="other DNA"
                     /db_xref="taxon:1750543"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      35..52
                     /regulatory_class="promoter"
                     /gene="SP6"
     gene            35..52
                     /gene="SP6"
                     /label=SP6
     CDS             103..1221
                     /codon_start=1
                     /gene="oCre"
                     /product="Cre recombinase"
                     /label=oCre
                     /note="fused witn NLS; optimized for Medaka (Oryzias 
                     latipes) codon usage"
                     /protein_id="BAT46644.1"
                     /translation="MAEALEASPPPKRPRLDSRMARPGGSGGTMSNLLTVHQNLPALPV
                     DATSDEVRKNLMDMFRDRQAFSEHTWKMLLSVCRSWAAWCKLNNRKWFPAEPEDVRDYL
                     LYLQARGLAVKTIQQHLGQLNMLHRRSGLPRPSDSNAVSLVMRRIRKENVDAGERAKQA
                     LAFERTDFDQVRSLMENSDRCQDIRNLAFLGIAYNTLLRIAEIARIRVKDISRTDGGRM
                     LIHIGRTKTLVSTAGVEKALSLGVTKLVERWISVSGVADDPNNYLFCRVRKNGVAAPSA
                     TSQLSTRALEGIFEATHRLIYGAKDDSGQRYLAWSGHSARVGAARDMARAGVSIPEIMQ
                     AGGWTNVNIVMNYIRNLDSETGAMVRLLEDGD"
     gene            103..1221
                     /gene="oCre"
                     /label=oCre
     misc_feature    127..153
                     /gene="oCre"
                     /note="NLS; PPPKRPRLD"
     promoter        complement(1241..1258)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1263..1397
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      1267..1458
                     /regulatory_class="polyA_signal_sequence"
                     /gene="SV40polyA"
                     /note="SV40 polyadenylation signal"
     gene            1267..1458
                     /gene="SV40polyA"
                     /label=SV40polyA
     promoter        complement(1510..1528)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1549..1565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1573..1589
                     /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(1597..1627)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1642..1663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1951..2539)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2710..3570)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3571..3675)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3701..4156)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4230..5214
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"