LOCUS       40924_44574        4804 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTY2b DNA, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4804)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Cell"; date: "2010"; volume: "22"; issue: "9"; pages: "3105-3117"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-JUL-2008) Contact:Hideya Fukuzawa Kyoto University, Graduate 
            School of Biostudies"; volume: " Kitashirakawa Oiwake-cho, Kyoto, 
            Kyoto 606-8502, Japan URL :http"; pages: 
            "//chlamy.pmb.lif.kyoto-u.ac.jp"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /label=AmpR promoter
     CDS             129..986
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1160..1748
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /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
                     /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
                     /note="includes enhancer region (-231 to -200 and -77 to
                     -25) of Chlamydomonas reinhardtii NAD(P)H nitrate reductase
                     protein (Nit1) gene"
                     /regulatory_class="enhancer"
     regulatory      2292..2375
                     /note="includes enhancer region (-231 to -200 and -77 to
                     -25) of Chlamydomonas reinhardtii NAD(P)H nitrate reductase
                     protein (Nit1) gene"
                     /regulatory_class="enhancer"
     regulatory      2376..2411
                     /note="includes promoter region (-36 to -1) of
                     Chlamydomonas reinhardtii beta2-tubulin (tubB2) gene"
                     /regulatory_class="promoter"
     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="3'UTR 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
                     /note="includes promoter region of Chlamydomonas
                     reinhardtii nuclear gene rbcS2 for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
                     /regulatory_class="promoter"
     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
     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"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctcgat ggtagggtgc
     2221 gagtgacccc gcgcgacttg gaagggttca aacgaccccg ccgtacgaac ttttgtcggg
     2281 gggcgctccc ggatggtagg gtgcgagtga ccccgcgcga cttggaaggg ttcaaacgac
     2341 cccgccgtac gaacttttgt cggggggcgc tcccgcgctg tttaaatagc caggcccccg
     2401 attgcaaaga cgctagcgaa gtcactcgag atcttaagct tgccaggtga gtcgacgagc
     2461 aagcccggcg gatcaggcag cgtgcttgca gatttgactt gcaacgcccg cattgtgtcg
     2521 acgaaggctt ttggctcctc tgtcgctgtc tcaagcagca tctaaccctg cgtcgccgtt
     2581 tccatttgca ggatggccat gcataccagc ggccaccgcg gtggcggccg ctctagaact
     2641 agtggatccc ccgggctgca ggaattcgat atcaagctta tcgatccccg ctccgtgtaa
     2701 atggaggcgc tcgttgatct gagccttgcc ccctgacgaa cggcggtgga tggaagatac
     2761 tgctctcaag tgctgaagcg gtagcttagc tccccgtttc gtgctgatca gtctttttca
     2821 acacgtaaaa agcggaggag ttttgcaatt ttgttggttg taacgatcct ccgttgattt
     2881 tggcctcttt ctccatgggc gggctgggcg tatttgaagc taaattaagc gatatcaagc
     2941 ttatcgatac cgtcgacctc gagatttaaa tgccagaagg agcgcagcca aaccaggatg
     3001 atgtttgatg gggtatttga gcacttgcaa cccttatccg gaagccccct ggcccacaaa
     3061 ggctaggcgc caatgcaagc agttcgcatg cagcccctgg agcggtgccc tcctgataaa
     3121 ccggccaggg ggcctatgtt ctttactttt ttacaagaga agtcactcaa catcttaaaa
     3181 tggccaggtg agtcgacgag caagcccggc ggatcaggca gcgtgcttgc agatttgact
     3241 tgcaacgccc gcattgtgtc gacgaaggct tttggctcct ctgtcgctgt ctcaagcagc
     3301 atctaaccct gcgtcgccgt ttccatttgc aggatggcca agctgaccag cgccgttccg
     3361 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     3421 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     3481 atcagcgcgg tccaggacca ggtgagtcga cgagcaagcc cggcggatca ggcagcgtgc
     3541 ttgcagattt gacttgcaac gcccgcattg tgtcgacgaa ggcttttggc tcctctgtcg
     3601 ctgtctcaag cagcatctaa ccctgcgtcg ccgtttccat ttgcaggacc aggtggtgcc
     3661 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
     3721 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
     3781 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
     3841 ggccgaggag caggactaac cgacgtcgac ccactctaga ggatcgatcc ccgctccgtg
     3901 taaatggagg cgctcgttga tctgagcctt gccccctgac gaacggcggt ggatggaaga
     3961 tactgctctc aagtgctgaa gcggtagctt agctccccgt ttcgtgctga tcagtctttt
     4021 tcaacacgta aaaagcggag gagttttgca attttgttgg ttgtaacgat cctccgttga
     4081 ttttggcctc tttctccatg ggcgggctgg gcgtatttga agcttaatta actcgagggg
     4141 gggcccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     4201 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     4261 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     4321 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     4381 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4441 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4501 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4561 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4621 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4681 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4741 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     4801 ttag
//
