pTop2 vector (V002571)

LOCUS       V002571                10829 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002571
VERSION     V002571
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10829)
  AUTHORS   Lu Y, Rijzaani H, Karcher D, Ruf S, Bock R.
  TITLE     Efficient engineering of the vitamin E metabolic pathway in
            transgenic tobacco and tomato plastids with synthetic multigene
            operons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10829)
  AUTHORS   Lu Y, Rijzaani H, Karcher D, Ruf S, Bock R.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2012) Organelle Biology, Biotechnology and
            Molecular Ecophysiology, Max Planck Institute of Molecular Plant
            Physiology, Am Muehlenberg 1, Potsdam-Golm, Brandenburg 14476,
            Germany
REFERENCE   3  (bases 1 to 10829)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10829)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (25-JUN-2012) Organelle Biology, Biotechnology and Molecular
            Ecophysiology, Max Planck Institute of Molecular Plant Physiology,
            Am Muehlenberg 1, Potsdam-Golm, Brandenburg 14476, Germany"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     tRNA            complement(31..104)
                     /product="tRNA-Met"
     CDS             complement(257..556)
                     /gene="rps14"
                     /label="Small ribosomal subunit protein uS14c"
                     /note="Small ribosomal subunit protein uS14c from Nicotiana
                     sylvestris. Accession#: Q3C1I0"
     gene            complement(679..1878)
                     /gene="psaB"
                     /label="psaB"
                     /note="derived from Nicotiana tabacum"
     promoter        complement(1891..1909)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1919..1935)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      2077..2532
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2558..2662
                     /label="AmpR promoter"
     CDS             2663..3520
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      3694..4282
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    4570..4591
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4606..4636
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4644..4660
                     /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     4668..4684
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        4705..4723
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     gene            4736..4867
                     /gene="psbZ"
                     /label="psbZ"
                     /note="derived from Nicotiana tabacum"
     tRNA            5143..5213
                     /product="tRNA-Gly"
     protein_bind    5406..5439
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      5480..5614
                     /note="prrN promoter; derived from Nicotiana tabacum"
                     /regulatory_class="promoter"
     CDS             5615..6403
                     /label="SmR"
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      6417..6811
                     /note="psbA terminator; derived from Nicotiana tabacum"
                     /regulatory_class="terminator"
     misc_recomb     6815..6848
                     /label="LoxP"
                     /note="LoxP"
     protein_bind    complement(6815..6848)
                     /label="loxP"
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT)."
     primer_bind     6878..6894
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      complement(6932..7153)
                     /note="rbcL terminator; derived from Chlamydomonas
                     reinhardtii"
                     /regulatory_class="terminator"
     CDS             complement(7160..8068)
                     /codon_start=1
                     /gene="TMT"
                     /product="gamma-tocopherol methyltransferase"
                     /label="TMT"
                     /note="derived from Arabidopsis thaliana"
                     /protein_id="AFQ99326.1"
                     /translation="MVAVAAAATSTEALRKGIAEFYNETSGLWEEIWGDHMHHGFYDPD
                     SSVQLSDSGHKEAQIRMIEESLRFAGVTDEEEEKKIKKVVDVGCGIGGSSRYLASKFGA
                     ECIGITLSPVQAKRANDLAAAQSLAHKASFQVADALDQPFEDGKFDLVWSMESGEHMPD
                     KAKFVKELVRVAAPGGRIIIVTWCHRNLSAGEEALQPWEQNILDKICKTFYLPAWCSTD
                     DYVNLLQSHSLQDIKCADWSENVAPFWPAVIRTALTWKGLVSLLRSGMKSIKGALTMPL
                     MIEGYKKGVIKFGIITCQKPL"
     gene            complement(7160..8068)
                     /gene="TMT"
                     /label="TMT"
                     /note="AtTMT"
     regulatory      8061..8070
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     stem_loop       complement(8088..8137)
                     /note="IEE; intercistronic expression element"
     regulatory      complement(8150..8299)
                     /note="rps16 terminator; derived from Nicotiana tabacum"
                     /regulatory_class="terminator"
     CDS             complement(8300..9391)
                     /codon_start=1
                     /gene="TCY"
                     /product="tocopherol cyclase"
                     /label="TCY"
                     /note="derived from Synechocystis sp. PCC 6803"
                     /protein_id="AFQ99327.1"
                     /translation="MKFPPHSGYHWQGQSPFFEGWYVRLLLPQSGESFAFMYSIENPAS
                     DHHYGGGAVQILGPATKKQENQEDQLVWRTFPSVKKFWASPRQFALGHWGKCRDNRQAK
                     PLLSEEFFATVKEGYQIHQNQHQGQIIHGDRHCRWQFTVEPEVTWGSPNRFPRATAGWL
                     SFLPLFDPGWQILLAQGRAHGWLKWQREQYEFDHALVYAEKNWGHSFPSRWFWLQANYF
                     PDHPGLSVTAAGGERIVLGRPEEVALIGLHHQGNFYEFGPGHGTVTWQVAPWGRWQLKA
                     SNDRYWVKLSGKTDKKGSLVHTPTAQGLQLNCRDTTRGYLYLQLGSVGHGLIVQGETDT
                     AGLEVGGDWGLTEENLSKKTVPF"
     gene            complement(8300..9391)
                     /gene="TCY"
                     /label="TCY"
                     /note="SyTCY"
     stem_loop       complement(8409..9458)
                     /note="IEE; intercistronic expression element"
     regulatory      complement(9464..9684)
                     /note="rbcL terminator; derived from Nicotiana tabacum"
                     /regulatory_class="terminator"
     CDS             complement(9688..10611)
                     /note="Homogentisate phytyltransferase from Synechocystis
                     sp. (strain PCC 6803 / Kazusa). Accession#: P73726"
                     /label="Homogentisate phytyltransferase"
     RBS             complement(10619..10641)
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(10776..10794)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"