LOCUS       Exported                3352 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pSH476, complete sequence.
ACCESSION   EU679367
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3352)
  AUTHORS   Hwang S-K., Okita TW.
  TITLE     Protein expression vector pSH476 (N-terminal His-tag)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3352)
  AUTHORS   Hwang S-K., Okita TW.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-APR-2008) Institute of Biological Chemistry, 
            Washington State University, Pullman, WA 99163, USA
REFERENCE   3  (bases 1 to 3352)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3352
                     /organism="Expression vector pSH476"
                     /mol_type="other DNA"
                     /db_xref="taxon:528210"
     regulatory      1..126
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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)."
     protein_bind    62..78
                     /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)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    127..150
                     /label=polyhistidine (His6)
                     /note="polyhistidine (His6)"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             151..885
                     /codon_start=1
                     /gene="mGFP"
                     /product="mGFP"
                     /label=mGFP
                     /note="green fluorescent protein; derived from pCAMBIA1302"
                     /protein_id="ACD50897.1"
                     /translation="MVDLTSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGK
                     LTLKFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFF
                     KDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKN
                     GIKANFKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHM
                     VLLEFVTAAGITHGMDELYKAS"
     gene            151..885
                     /gene="mGFP"
                     /label=mGFP
     CDS             166..876
                     /codon_start=1
                     /gene="mgfp5"
                     /product="GFP with folding enhancement mutations"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
                     FKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     regulatory      951..1000
                     /regulatory_class="terminator"
                     /note="lambda t0"
     terminator      955..1049
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    1202..1342
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1340..1440)
                     /direction=LEFT
                     /label=Ori
                     /note="Ori"
     rep_origin      complement(1528..2116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2287..3147)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(2287..2952)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /protein_id="ACD50898.1"
                     /translation="MMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKH
                     LTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNE
                     AIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWF
                     IADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     promoter        complement(3148..3252)
                     /gene="bla"
                     /label=AmpR promoter