LOCUS       Exported                3342 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pME-LifeAct-GFP, complete sequence.
ACCESSION   JN717248
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3342)
  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 3342)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3342
                     /organism="Cloning vector pME-LifeAct-GFP"
                     /mol_type="other DNA"
                     /label=lifeAct-GFP in pDONR 221
                     /note="lifeAct-GFP in pDONR 221"
                     /db_xref="taxon:1115844"
     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..745
                     /label=LifeAct 5-prime tag
                     /note="LifeAct 5-prime tag"
     CDS             674..724
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     regulatory      740..749
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             746..1465
                     /codon_start=1
                     /product="EGFP"
                     /label=EGFP
                     /protein_id="AET83985.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    complement(1467..1566)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     complement(1470..1565)
                     /label=AttL2
                     /note="AttL2"
     misc_recomb     complement(1470..1490)
                     /label=attB2
                     /note="attB2"
     primer_bind     complement(1583..1604)
                     /label=T7
                     /note="T7"
     promoter        complement(1584..1602)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1603..1623)
                     /label=M13-rev
                     /note="M13-rev"
     primer_bind     complement(1607..1623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1736..2545
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AET83984.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2688..3295
                     /label=pBR322 Origin
                     /note="pBR322 Origin"
     rep_origin      2692..3280
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"