LOCUS       Exported                6242 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pTha-M1 DNA, complete sequence.
ACCESSION   AB849919
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6242)
  AUTHORS   Tanaka R, Kikutani S, Mahardika A, Matsuda Y.
  TITLE     Localization of enzymes relating to C4 organic acid metabolisms in 
            the marine diatom, Thalassiosira pseudonana
  JOURNAL   Photosyn. Res. 121 (2-3), 251-263 (2014)
  PUBMED    24414292
REFERENCE   2  (bases 1 to 6242)
  AUTHORS   Kikutani S, Matsuda Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-AUG-2013) Contact:Sae Kikutani Kwansei Gakuin 
            University, Department of Bioscience; 2-1 Gakuen, Sanda, Hyogo 
            669-1337, Japan
REFERENCE   3  (bases 1 to 6242)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6242
                     /organism="Shuttle vector pTha-M1"
                     /lab_host="Thalassiosira pseudonana"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:1395175"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /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    659..700
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             701..1420
                     /codon_start=1
                     /product="EGFP"
                     /label=EGFP
                     /protein_id="BAO52747.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      1429..1931
                     /regulatory_class="terminator"
                     /note="Tpfcp8"
     regulatory      1958..2916
                     /regulatory_class="promoter"
                     /note="Tpfcp8"
     CDS             2924..3493
                     /codon_start=1
                     /product="nourseothricin N-acetyl-transferase"
                     /function="Nourseothricin resistance"
                     /label=nourseothricin N-acetyl-transferase
                     /protein_id="BAO52748.1"
                     /translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
                     LREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRR
                     LTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTL
                     CGLDTALYDGTASDGEQALYMSMPCP"
     regulatory      3493..3996
                     /regulatory_class="terminator"
                     /note="Tpfcp8"
     primer_bind     complement(3997..4013)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4054..4072)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4093..4109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4117..4133
                     /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(4141..4171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4186..4207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4495..5083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5254..6114)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="BAO52749.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6115..6219)
                     /gene="bla"
                     /label=AmpR promoter