LOCUS       Exported                6084 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEF-F13F14-tdTomato-dpuro, complete sequence.
ACCESSION   KT351863
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6084)
  AUTHORS   Baser B, Spehr J, Bussow K, van den Heuvel J.
  TITLE     A method for specifically targeting two independent genomic 
            integration sites for co-expression of genes in CHO cells
  JOURNAL   Methods (2015) In press
  PUBMED    26658354
REFERENCE   2  (bases 1 to 6084)
  AUTHORS   Baser B, Spehr J, Buessow K, van den Heuvel J.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUL-2015) Structure and Function of Proteins, 
            Recombinant Protein Expression, Helmholtz Centre for Infection 
            Research, Inhoffenstrasse 7, Braunschweig, Lower Saxony 38124, 
            Germany
REFERENCE   3  (bases 1 to 6084)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6084
                     /organism="Cloning vector pEF-F13F14-tdTomato-dpuro"
                     /mol_type="other DNA"
                     /db_xref="taxon:1770594"
     regulatory      111..1300
                     /regulatory_class="promoter"
                     /note="derived from Homo sapiens 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="FRT13; synthetic variant 13 of the yeast 
                     flp-recombinase target (FRT)"
     regulatory      1382..1391
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1388..2836
                     /codon_start=1
                     /product="td-Tomato-His6"
                     /label=td-Tomato-His6
                     /note="tandem Tomato fluorescence protein with 
                     hexahistidine-tag"
                     /protein_id="ALT10469.1"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYKHHHHHH"
     CDS             1388..2815
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             2816..2833
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2851..2985
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2855..3076
                     /regulatory_class="polyA_signal_sequence"
                     /note="derived from SV40 virus"
     misc_recomb     3098..3145
                     /note="FRT14; synthetic variant 14 of the yeast 
                     flp-recombinase target (FRT)"
     misc_feature    3165..3761
                     /note="similar to puromycin-N-acetyl-transferase; puromycin
                     resistance gene; inactive selection trap"
     CDS             3165..3761
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(3805..3823)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      3849..4073
                     /regulatory_class="polyA_signal_sequence"
                     /note="derived from bovine growth hormone gene (BGH)"
     polyA_signal    3849..4073
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4273..4946
                     /label=pUC origin of replication
                     /note="pUC origin of replication"
     rep_origin      complement(4332..4920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5091..5951)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase precursor"
                     /EC_number="3.5.2.6"
                     /label=bla
                     /note="provides resistance to beta-lactam antibiotics in 
                     prokaryotes; similar to the beta-lactamase precursor 
                     encoded by INSDC accession number AF074376; penicillinase 
                     precursor"
                     /protein_id="ALT10470.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5091..5951)
                     /gene="bla"
                     /label=bla
     promoter        complement(5952..6056)
                     /gene="bla"
                     /label=AmpR promoter