LOCUS       Exported                6572 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pAttR-YFP-2xHA, complete sequence.
ACCESSION   HM989367
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6572)
  AUTHORS   Conn SJ.
  TITLE     Magnesium transporters in Arabidopsis thaliana
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6572)
  AUTHORS   Conn SJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-JUL-2010) Agriculture, Food & Wine, University of 
            Adelaide, PMB1, Glen Osmond, SA 5064, Australia
REFERENCE   3  (bases 1 to 6572)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6572
                     /organism="Expression vector pAttR-YFP-2xHA"
                     /mol_type="other DNA"
                     /db_xref="taxon:934313"
     regulatory      1..750
                     /regulatory_class="promoter"
                     /note="35S cauliflower mosaic virus promoter"
     promoter        412..756
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     regulatory      756..898
                     /regulatory_class="enhancer"
                     /note="translational enhancer; 5 UTR from tobacco etch 
                     virus"
     misc_recomb     904..1028
                     /label=AttR1
                     /note="AttR1"
     protein_bind    904..1028
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1053..1083
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1137..1796
                     /codon_start=1
                     /product="chloramphenicol resistance"
                     /label=chloramphenicol resistance
                     /protein_id="ADU18045.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2138..2443
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /note="encodes a cytotoxic protein that is a potent poison 
                     of DNA gyrase"
                     /protein_id="ADU18046.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            2138..2443
                     /gene="ccdB"
                     /label=ccdB
     misc_recomb     2484..2608
                     /label=AttR2
                     /note="AttR2"
     protein_bind    complement(2484..2608)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2610..3410
                     /codon_start=1
                     /product="YFP-haemagglutinin fusion protein"
                     /label=YFP-haemagglutinin fusion protein
                     /note="yellow fluorescent protein; 2xHA"
                     /protein_id="ADU18044.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKRSIYPYDVPDYARSIYPYDVPDYARSI"
     misc_feature    2610..3335
                     /label=YFP
                     /note="YFP"
     CDS             2610..3326
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    3336..3410
                     /note="haemagglutinin; 2xHA"
     CDS             3336..3362
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3372..3398
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3683..3701)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3722..3738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3746..3762
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3770..3800)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3815..3836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4124..4712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      4228..4508
                     /label=pBR322 ori
                     /note="pBR322 ori"
     CDS             complement(4883..5743)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /note="confers resistance to ampicillin antibiotic"
                     /protein_id="ADU18047.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5744..5848)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5874..6329)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6471..6487
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6494..6512
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     6537..6553
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"