LOCUS       Exported                6461 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pTha-K1 DNA, complete sequence.
ACCESSION   AB849918
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6461)
  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 6461)
  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 6461)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6461
                     /organism="Shuttle vector pTha-K1"
                     /lab_host="Thalassiosira pseudonana"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:1395174"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      58..1023
                     /regulatory_class="promoter"
                     /note="Tpfcp8"
     CDS             1024..1593
                     /codon_start=1
                     /product="nourseothricin N-acetyl-transferase"
                     /function="Nourseothricin resistance"
                     /label=nourseothricin N-acetyl-transferase
                     /note="Nat1"
                     /protein_id="BAO52745.1"
                     /translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
                     LREVPVDPPLTKVFPDDESDDESDAGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRR
                     LTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTL
                     CGLDTALYDGTASDGEQALYMSMPCP"
     regulatory      1594..2096
                     /regulatory_class="terminator"
                     /note="Tpfcp8"
     regulatory      2131..3145
                     /regulatory_class="promoter"
                     /note="Tpfcp8"
     misc_feature    3146..3165
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      3166..3668
                     /regulatory_class="terminator"
                     /note="Tpfcp8"
     primer_bind     complement(3700..3716)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3858..4313)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4340..4444
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4445..5305
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="BAO52746.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5476..6064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6352..6373
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6388..6418
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6426..6442
                     /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     6450..5
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"