LOCUS       Exported                4107 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pME-mCherry-T2A-EGFP, complete sequence.
ACCESSION   JN717245
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4107)
  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 4107)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4107
                     /organism="Cloning vector pME-mCherry-T2A-EGFP"
                     /mol_type="other DNA"
                     /label=mCherry-T2A-EGFP in pDONR 221
                     /note="mCherry-T2A-EGFP in pDONR 221"
                     /db_xref="taxon:1115841"
     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    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     570..665
                     /label=AttL1
                     /note="AttL1"
     misc_recomb     645..665
                     /label=attB1
                     /note="attB1"
     misc_feature    674..1381
                     /label=similar to mCherry
                     /note="similar to mCherry"
     CDS             674..1381
                     /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    1382..1435
                     /label=T2A viral peptide
                     /note="T2A viral peptide"
     CDS             1382..1435
                     /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    1436..2152
                     /label=similar to EGFP
                     /note="similar to EGFP"
     CDS             1436..2152
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    2168..2230
                     /label=CAAX membrane localisation signal (from H-ras)
                     /note="CAAX membrane localisation signal (from H-ras)"
     protein_bind    complement(2232..2331)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     complement(2235..2330)
                     /label=AttL2
                     /note="AttL2"
     misc_recomb     complement(2235..2255)
                     /label=attB2
                     /note="attB2"
     primer_bind     complement(2348..2369)
                     /label=T7
                     /note="T7"
     promoter        complement(2349..2367)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2368..2388)
                     /label=M13-rev
                     /note="M13-rev"
     primer_bind     complement(2372..2388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2501..3310
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AET83980.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3453..4060
                     /label=pBR322 Origin
                     /note="pBR322 Origin"
     rep_origin      3457..4045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"