LOCUS       Exported                7570 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCMV-Cre-IRES-eGFP, complete sequence.
ACCESSION   MG550106
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7570)
  AUTHORS   Kozhevnikova EN, Leshchenko AE, Pindyurin AV.
  TITLE     An inducible DamID system for profiling of a nuclear lamina protein 
            component Lamin B1 interactions with chromosomes in mouse cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7570)
  AUTHORS   Kozhevnikova EN, Leshchenko AE, Pindyurin AV.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-NOV-2017) Department of the Regulation of Genetic 
            Processes, Institute of Molecular and Cellular Biology, Lavrentyev 
            ave. 8/2, Novosibirsk, Novosibirsk Region 630090, Russia
REFERENCE   3  (bases 1 to 7570)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7570
                     /organism="Cloning vector pCMV-Cre-IRES-eGFP"
                     /mol_type="other DNA"
                     /label=Cre recombinase expression vector
                     /note="Cre recombinase expression vector"
                     /db_xref="taxon:2066509"
     regulatory      100..675
                     /regulatory_class="promoter"
                     /note="human cytomegalovirus immediate early gene 
                     enhancer/promoter; CMV promoter"
     enhancer        100..403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        404..607
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             715..1770
                     /codon_start=1
                     /product="Cre recombinase"
                     /label=Cre recombinase
                     /note="contains nuclear localization signal at the 
                     N-terminus"
                     /protein_id="AUM57422.1"
                     /translation="MAPKKKRKVSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAF
                     SEHTWKMLLSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLN
                     MLHRRSGLPRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRC
                     QDIRNLAFLGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALS
                     LGVTKLVERWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIY
                     GAKDDSGQRYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSET
                     GAMVRLLEDGD"
     CDS             721..741
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             742..1770
                     /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
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     regulatory      1865..2451
                     /regulatory_class="transcriptional_cis_regulatory_region"
                     /note="IRES; encephalomyocarditis virus internal ribosomal 
                     entry site"
     misc_feature    1865..2451
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2452..3171
                     /codon_start=1
                     /product="enhanced green fluorescent protein"
                     /label=enhanced green fluorescent protein
                     /note="eGFP"
                     /protein_id="AUM57423.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3294..3415
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    3418..4010
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element; WPRE"
     misc_feature    3422..4010
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3893..3904)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      4095..4330
                     /regulatory_class="terminator"
                     /note="HIV-1 self-inactivating 3 long terminal repeat; 3 
                     LTR delta-U3"
     LTR             4097..4330
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4402..4523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      4563..4698
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(4719..4737)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4747..4763)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4905..5360
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5386..5490
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5491..6351
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance protein"
                     /protein_id="AUM57424.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            5491..6351
                     /gene="bla"
                     /label=bla
     rep_origin      6522..7110
                     /note="high-copy-number; ColE1/pMB1/pBR322/pUC"
     protein_bind    7398..7419
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7434..7464
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7472..7488
                     /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)."
     primer_bind     7496..7512
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7533..7551
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"