LOCUS       Exported                4401 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p3E-mCherry-T2A-EGFPcaax, complete sequence.
ACCESSION   JN717246
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4401)
  AUTHORS   Hall TE, Currie PD.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-SEP-2011) Australian Regenerative Medicine Institute, 
            Monash University, Building 75, Melbourne, VIC 3800, Australia
REFERENCE   2  (bases 1 to 4401)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4401
                     /organism="Cloning vector p3E-mCherry-T2A-EGFPcaax"
                     /mol_type="other DNA"
                     /label=mCherry-T2A-EGFPcaax in pDONR p2R-p3
                     /note="mCherry-T2A-EGFPcaax in pDONR p2R-p3"
                     /db_xref="taxon:1115842"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     536..553
                     /label=M13-fwd
                     /note="M13-fwd"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(624..748)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     complement(727..747)
                     /label=attB2
                     /note="attB2"
     misc_feature    756..1463
                     /label=similar to mCherry
                     /note="similar to mCherry"
     CDS             756..1463
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    1464..1517
                     /label=T2A viral peptide
                     /note="T2A viral peptide"
     CDS             1464..1517
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    1518..2234
                     /label=similar to EGFP
                     /note="similar to EGFP"
     CDS             1518..2234
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    2250..2312
                     /label=CAAX membrane tag (H-ras)
                     /note="CAAX membrane tag (H-ras)"
     polyA_signal    2313..2447
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2317..2508
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 polyA"
     misc_recomb     complement(2510..2605)
                     /label=attL3
                     /note="attL3"
     protein_bind    complement(2510..2605)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     complement(2510..2530)
                     /label=attB3
                     /note="attB3"
     primer_bind     complement(2642..2663)
                     /label=T7
                     /note="T7"
     promoter        complement(2643..2661)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2662..2682)
                     /label=M13-rev
                     /note="M13-rev"
     primer_bind     complement(2666..2682)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2795..3604
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AET83981.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    3725..4398
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      3751..4339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"