LOCUS       Exported                5325 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHA1001-ble-PeEGFP DNA, complete sequence.
ACCESSION   LC184596
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5325)
  AUTHORS   Abe J, Harayama S.
  TITLE     Expression of ble-PeEGFP fusion protein for bleomycin resistance 
            protein and codon-optimized green fluorescence protein in 
            Pseudococcomyxa sp. KJ
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5325)
  AUTHORS   Abe J, Harayama S.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-SEP-2016) Contact:Jun Abe Chuo University, Faculty of 
            Science and Engineering; 1-13-27 Kasuga, Bunkyo-ku, Tokyo 112-8551, 
            Japan
REFERENCE   3  (bases 1 to 5325)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5325
                     /organism="Expression vector pHA1001-ble-PeEGFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1899313"
     misc_feature    1..2237
                     /label=derived from plasmid pBluescript SK(+)
                     /note="derived from plasmid pBluescript SK(+)"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /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"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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)."
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      2238..3027
                     /regulatory_class="promoter"
                     /note="includes promoter region of Pseudococcomyxa 
                     ellipsoidea nuclear gene RBCS for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
     CDS             3028..4134
                     /codon_start=1
                     /gene="ble-PeEGFP"
                     /product="bleomycin resistance protein/codon-optimized 
                     green fluorescence protein fusion protein"
                     /label=ble-PeEGFP
                     /protein_id="BAV58188.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQDGGSGGRMAKGEELFTGVVPILVELDGDVNGHKFSVSGEG
                     EGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGY
                     VQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVY
                     IMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKD
                     PNEKRDHMVLLEFVTAAGITLGMDELYK"
     gene            3028..4134
                     /gene="ble-PeEGFP"
                     /label=ble-PeEGFP
     misc_feature    3028..3399
                     /gene="ble-PeEGFP"
                     /note="ble; bleomycin resistance protein"
     misc_feature    3400..3417
                     /gene="ble-PeEGFP"
                     /label=linker
                     /note="linker"
     misc_feature    3418..4134
                     /gene="ble-PeEGFP"
                     /note="PeEGFP; codon-optimized green fluorescence protein"
     regulatory      4141..4652
                     /regulatory_class="terminator"
                     /note="includes terminator region of Pseudococcomyxa 
                     ellipsoidea RBCS gene"
     misc_feature    4653..5325
                     /label=derived from plasmid pBluescript SK(+)
                     /note="derived from plasmid pBluescript SK(+)"
     promoter        complement(4682..4700)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4710..4726)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4867..5322)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"