LOCUS       Exported                4835 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pBS-35S-Ala-YFP, complete sequence.
ACCESSION   AY189984
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4835)
  AUTHORS   von Arnim A.
  TITLE     Vectors for Bioluminescence Resonance Energy Transfer
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4835)
  AUTHORS   von Arnim A.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-DEC-2002) Botany, University of Tennessee, 1406 Circle
            Drive, Knoxville, TN 37996, USA
REFERENCE   3  (bases 1 to 4835)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4835
                     /organism="Expression vector pBS-35S-Ala-YFP"
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:221063"
     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"
     misc_feature    653..704
                     /label=polylinker
                     /note="polylinker"
     primer_bind     669..685
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      705..1459
                     /regulatory_class="promoter"
                     /note="cauliflower mosaic virus dual 35S promoter"
     promoter        1116..1460
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     5UTR           1460..1602
                     /note="translational enhancer; 5UTR from tobacco etch 
                     virus"
     regulatory      1601..1610
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1603..2373
                     /codon_start=1
                     /gene="YFP-AlaN"
                     /product="yellow fluorescent protein"
                     /label=YFP-AlaN
                     /note="YFP reporter protein; contains a N-terminal alanine 
                     linker"
                     /protein_id="AAO48597.1"
                     /translation="MAAAAPVAAAAAALMVSKGEELFTGVVPILVELDGDVNGHKFSVS
                     GEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMP
                     EGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSH
                     NVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSAL
                     SKDPNEKRDHMVLLEFVTAAGITLGMDELYKRSI"
     gene            1603..2373
                     /gene="YFP-AlaN"
                     /label=YFP-AlaN
     CDS             1645..2361
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2374..2584
                     /regulatory_class="terminator"
                     /note="cauliflower mosaic virus 35S terminator"
     promoter        complement(2650..2668)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2689..2705)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2713..2729
                     /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(2737..2767)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2782..2803
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3091..3679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3850..4710)
                     /codon_start=1
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=beta-lactamase
                     /protein_id="AAO48598.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4711..4815)
                     /gene="bla"
                     /label=AmpR promoter