LOCUS       Exported                6500 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBS 35S attR-YFP, complete sequence.
ACCESSION   AY995141
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6500)
  AUTHORS   Subramanian C, Woo J, Cai X, Xu X, Servick S, Johnson CH, Nebenfuhr 
            A, von Arnim AG.
  TITLE     A suite of tools and application notes for in vivo protein 
            interaction assays using bioluminescence resonance energy transfer 
            (BRET)
  JOURNAL   Plant J. 48 (1), 138-152 (2006)
  PUBMED    16925598
REFERENCE   2  (bases 1 to 6500)
  AUTHORS   von Arnim AG, Subramanian C.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-MAR-2005) BCMB, University of Tennessee Knoxville, 
            1406 Circle Drive, Hesler Biology Bldg 618, Knoxville, TN 37996, USA
REFERENCE   3  (bases 1 to 6500)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6500
                     /organism="Cloning vector pBS 35S attR-YFP"
                     /mol_type="other DNA"
                     /note="plant gateway cloning vector for bioluminescence 
                     resonance energy transfer"
                     /db_xref="taxon:324064"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     669..685
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      705..1602
                     /regulatory_class="promoter"
                     /note="35S cauliflower mosaic virus promoter"
     promoter        1116..1460
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     regulatory      1460..1602
                     /regulatory_class="enhancer"
                     /note="translational enhancer; 5 UTR from tobacco etch 
                     virus"
     misc_feature    1608..1732
                     /label=attR1 gateway recombination site
                     /note="attR1 gateway recombination site"
     protein_bind    1608..1732
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1757..1787
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1841..2500
                     /codon_start=1
                     /product="CmR"
                     /label=CmR
                     /note="confers resistance to antibiotic chloramphenicol"
                     /protein_id="AAX97753.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2842..3147
                     /codon_start=1
                     /product="ccdB"
                     /label=ccdB
                     /note="encodes a cytotoxic protein that is a potent poison 
                     of DNA gyrase"
                     /protein_id="AAX97754.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    3188..3312
                     /label=attR2 gateway recombination site
                     /note="attR2 gateway recombination site"
     protein_bind    complement(3188..3312)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3314..4042
                     /codon_start=1
                     /product="EYFP"
                     /label=EYFP
                     /note="yellow fluorescent protein reporter"
                     /protein_id="AAX97752.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKRSI"
     CDS             3314..4030
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      4043..4253
                     /regulatory_class="terminator"
                     /note="35S cauliflower mosaic virus terminator"
     promoter        complement(4315..4333)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4354..4370)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4378..4394
                     /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(4402..4432)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4447..4468
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4756..5344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5515..6375)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="confers resistance to antibiotic ampicillin"
                     /protein_id="AAX97755.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6376..6480)
                     /gene="bla"
                     /label=AmpR promoter