LOCUS       Exported                4804 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTY2b DNA, complete sequence.
ACCESSION   AB447355
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4804)
  AUTHORS   Ohnishi N, Mukherjee B, Tsujikawa T, Yanase M, Nakano H, Moroney JV,
            Fukuzawa H.
  TITLE     Expression of a low CO-inducible protein, LCI1, increases inorganic 
            carbon uptake in the green alga Chlamydomonas reinhardtii
  JOURNAL   Plant Cell 22 (9), 3105-3117 (2010)
  PUBMED    20870960
REFERENCE   2  (bases 1 to 4804)
  AUTHORS   Tsujikawa T, Fukuzawa H.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUL-2008) Contact:Hideya Fukuzawa Kyoto University, 
            Graduate School of Biostudies; Kitashirakawa Oiwake-cho, Kyoto, 
            Kyoto 606-8502, Japan URL :http://chlamy.pmb.lif.kyoto-u.ac.jp/
REFERENCE   3  (bases 1 to 4804)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4804
                     /organism="Expression vector pTY2b"
                     /mol_type="other DNA"
                     /db_xref="taxon:547989"
     misc_feature    1..2201
                     /note="derived from plasmid pBluescript SK(+), Genbank 
                     accession number X52328"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /function="confers resistance to ampicilin"
                     /label=bla
                     /protein_id="BAI39423.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            129..989
                     /gene="bla"
                     /label=bla
     rep_origin      1137..1804
                     /label=pUC origin
                     /note="pUC origin"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      2024..2145
                     /regulatory_class="promoter"
                     /gene="lacZ"
                     /note="T7"
     gene            2024..2145
                     /gene="lacZ"
                     /label=lacZ
     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      2208..2291
                     /regulatory_class="enhancer"
                     /note="includes enhancer region (-231 to -200 and -77 to 
                     -25) of Chlamydomonas reinhardtii NAD(P)H nitrate reductase
                     protein (Nit1) gene"
     regulatory      2292..2375
                     /regulatory_class="enhancer"
                     /note="includes enhancer region (-231 to -200 and -77 to 
                     -25) of Chlamydomonas reinhardtii NAD(P)H nitrate reductase
                     protein (Nit1) gene"
     regulatory      2376..2411
                     /regulatory_class="promoter"
                     /note="includes promoter region (-36 to -1) of 
                     Chlamydomonas reinhardtii beta2-tubulin (tubB2) gene"
     misc_feature    2418..2612
                     /note="includes intron 1 of Chlamydomonas reinhardtii 
                     nuclear gene rbcS2 for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
     primer_bind     2632..2648
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2680..2863
                     /note="3UTR region of Chlamydomonas reinhardtii nuclear 
                     gene rbcS2 for ribulose bisphosphate carboxylase oxygenase 
                     small subunit"
     primer_bind     complement(2945..2961)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2974..3188
                     /regulatory_class="promoter"
                     /note="includes promoter region of Chlamydomonas 
                     reinhardtii nuclear gene rbcS2 for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
     misc_feature    3188..3333
                     /note="includes intron 1 from Chlamydomonas reinhardtii 
                     rbcS2 gene"
     CDS             join(3334..3501,3647..3859)
                     /codon_start=1
                     /gene="ble"
                     /product="bleomycin resistance protein"
                     /label=ble
                     /note="derived from Streptoalloteichus hindustanus; fusion 
                     zeocin binding protein; confers resistance to zeocin"
                     /protein_id="BAI39424.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPW
                     GREFALRDPAGNCVHFVAEEQD"
     misc_feature    3334..3501
                     /gene="ble"
                     /note="includes a fragment of Streptoalloteichus 
                     hindustanus Ble gene"
     gene            3334..3501
                     /gene="ble"
                     /label=ble
     misc_feature    3502..3646
                     /note="includes intron 1 of Chlamydomonas reinhardtii rbcS2
                     gene"
     misc_feature    3647..3859
                     /gene="ble"
                     /note="includes a fragment of Streptoalloteichus 
                     hindustanus Ble gene and stop codon"
     gene            3647..3859
                     /gene="ble"
                     /label=ble
     misc_feature    3860..4804
                     /note="derived from plasmid pBluescript SK(+), Genbank 
                     accession number X52328"
     promoter        complement(4161..4179)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4189..4205)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4347..4802
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      4361..4667
                     /label=f1 origin
                     /note="f1 origin"