LOCUS       Exported                4766 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEF-F3-EGFP-F3-mcs, complete sequence.
ACCESSION   GU983383
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4766)
  AUTHORS   Kaufman WL, Kocman I, Agrawal V, Rahn HP, Besser D, Gossen M.
  TITLE     Homogeneity and persistence of transgene expression by omitting 
            antibiotic selection in cell line isolation
  JOURNAL   Nucleic Acids Res. 36 (17), E111 (2008)
  PUBMED    18682524
REFERENCE   2  (bases 1 to 4766)
  AUTHORS   Kaufman WL, Gossen M.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-MAR-2010) Structural Biology, Helmholtz Centre for 
            Infection Research, Inhoffenstr. 7, Braunschweig 38124, Germany
REFERENCE   3  (bases 1 to 4766)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4766
                     /organism="Cloning vector pEF-F3-EGFP-F3-mcs"
                     /mol_type="other DNA"
                     /db_xref="taxon:747266"
     regulatory      111..1300
                     /regulatory_class="promoter"
                     /note="human elongation factor 1 alpha"
     promoter        111..1300
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          341..1291
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     misc_recomb     1322..1369
                     /note="FRT-3 site; mutant 3 of the flp-recombinase target 
                     (FRT), the target site of yeast flp recombinase (flippase)"
     regulatory      1382..1391
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1388..2134
                     /codon_start=1
                     /gene="gfp"
                     /product="green fluorescent protein"
                     /label=gfp
                     /note="GFP"
                     /protein_id="ADD98904.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKSGRTQIHRI"
     gene            1388..2134
                     /gene="gfp"
                     /label=gfp
     CDS             1388..2104
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2247..2368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2278..2312
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40"
     primer_bind     2322..2339
                     /label=PCR primer pEFGM
                     /note="PCR primer pEFGM"
     misc_recomb     2375..2422
                     /note="FRT-3 site; mutant 3 of the flp-recombinase target 
                     (FRT), the target site of yeast flp recombinase (flippase)"
     misc_feature    2425..2530
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     complement(2436..2452)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2487..2505)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2525..2542)
                     /label=PCR primer BGHrev
                     /note="PCR primer BGHrev"
     regulatory      2531..2755
                     /regulatory_class="polyA_signal_sequence"
                     /note="bovine growth hormone (BGH)"
     polyA_signal    2531..2755
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2955..3628
                     /label=pUC
                     /note="pUC"
     rep_origin      complement(3014..3602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3773..4633)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase precursor"
                     /EC_number="3.5.2.6"
                     /label=bla
                     /note="penicillinase; similar to the protein encoded by 
                     INSD accession AF074376"
                     /protein_id="ADD98905.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3773..4633)
                     /gene="bla"
                     /label=bla
     promoter        complement(4634..4738)
                     /gene="bla"
                     /label=AmpR promoter