LOCUS       Exported                4410 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pME-h2afv-EosFPtd, complete sequence.
ACCESSION   JN717247
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4410)
  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 4410)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4410
                     /organism="Cloning vector pME-h2afv-EosFPtd"
                     /mol_type="other DNA"
                     /label=nuclear localised EosFPtd in pDONR 221
                     /note="nuclear localised EosFPtd in pDONR 221"
                     /db_xref="taxon:1115843"
     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"
     regulatory      668..677
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             674..2524
                     /codon_start=1
                     /product="h2afv/EosFPtd fusion protein"
                     /label=h2afv/EosFPtd fusion protein
                     /protein_id="AET83983.1"
                     /translation="MAGGKAGKDSGKAKAKAVSRSQRAGLQFPVGRIHRHLKTRTTSHG
                     RVGATAAVYSAAILEYLTAEVLELAGNASKDLKVKRITPRHLQLAIRGDEELDSLIKAT
                     IAGGGVIPHIHKSLIGKKGQQKTAGGMDYKDDDDKEFCRYPSHWRPLESAIKPDMKINL
                     RMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKEGGPLPFAFDILTTAFHYGNRVFAEY
                     PDHIQDYFKQSFPKGYSWERSLTFEDGGICIARNDITMEGDTFYNKVRFHGVNFPANGP
                     VMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLLEGNAHYRCDFRTTYKAKEKGVKLPGY
                     HFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDNARRSRGHGTGSTGSGSSSAIKPDMK
                     INLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKEGGPLPFAFDILTTAFHYGNRVF
                     AEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIARNDITMEGDTFYNKVRFHGVNFPA
                     NGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLLEGNAHYRCDFRTTYKAKEKGVKL
                     PGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDNARR"
     misc_feature    1064..1090
                     /label=Region: FLAG tag
                     /note="Region: FLAG tag"
     CDS             1067..1090
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1130..1804
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="SAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIARN
                     DITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     CDS             1847..2524
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="SAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIARN
                     DITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     protein_bind    complement(2535..2634)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     complement(2538..2633)
                     /label=AttL2
                     /note="AttL2"
     misc_recomb     complement(2538..2558)
                     /label=attB2
                     /note="attB2"
     primer_bind     complement(2651..2672)
                     /label=T7
                     /note="T7"
     promoter        complement(2652..2670)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2671..2691)
                     /label=M13-rev
                     /note="M13-rev"
     primer_bind     complement(2675..2691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2804..3613
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AET83982.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3756..4363
                     /label=pBR322
                     /note="pBR322"
     rep_origin      3760..4348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"