Basic Vector Information
- Vector Name:
- pTop1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10745 bp
- Type:
- Chloroplast transformation vector
- Replication origin:
- ori
- Source/Author:
- Lu Y, Rijzaani H, Karcher D, Ruf S, Bock R.
- Promoter:
- T3
pTop1 vector Map
pTop1 vector Sequence
LOCUS V002572 10745 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V002572
VERSION V002572
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10745)
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JOURNAL Unpublished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 10745)
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 10745)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10745)
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"Unpublished"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..10745
/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(6897..7117)
/note="rbcL terminator; derived from Nicotiana tabacum"
/regulatory_class="terminator"
CDS complement(7121..8044)
/note="Homogentisate phytyltransferase from Synechocystis
sp. (strain PCC 6803 / Kazusa). Accession#: P73726"
/label="Homogentisate phytyltransferase"
misc_feature complement(8045..8291)
/note="IGR2; intergenic region between Nicotiana tabacum
plastid rps2 and atpI"
CDS complement(8324..9415)
/codon_start=1
/gene="TCY"
/product="tocopherol cyclase"
/label="TCY"
/note="derived from Synechocystis sp. PCC 6803"
/protein_id="AFQ99321.1"
/translation="MKFPPHSGYHWQGQSPFFEGWYVRLLLPQSGESFAFMYSIENPAS
DHHYGGGAVQILGPATKKQENQEDQLVWRTFPSVKKFWASPRQFALGHWGKCRDNRQAK
PLLSEEFFATVKEGYQIHQNQHQGQIIHGDRHCRWQFTVEPEVTWGSPNRFPRATAGWL
SFLPLFDPGWQILLAQGRAHGWLKWQREQYEFDHALVYAEKNWGHSFPSRWFWLQANYF
PDHPGLSVTAAGGERIVLGRPEEVALIGLHHQGNFYEFGPGHGTVTWQVAPWGRWQLKA
SNDRYWVKLSGKTDKKGSLVHTPTAQGLQLNCRDTTRGYLYLQLGSVGHGLIVQGETDT
AGLEVGGDWGLTEENLSKKTVPF"
gene complement(8324..9415)
/gene="TCY"
/label="TCY"
/note="SyTCY"
misc_feature complement(9416..9564)
/note="IGR1; intergenic region between Nicotiana tabacum
plastid psbH and petB"
CDS complement(9574..10527)
/codon_start=1
/gene="TMT"
/product="gamma-tocopherol methyltransferase"
/label="TMT"
/note="derived from Synechocystis sp. PCC 6803"
/protein_id="AFQ99322.1"
/translation="MVYHVRPKHALFLAFYCYFSLLTMASATIASADLYEKIKNFYDDS
SGLWEDVWGEHMHHGYYGPHGTYRIDRRQAQIDLIKELLAWAVPQNSAKPRKILDLGCG
IGGSSLYLAQQHQAEVMGASLSPVQVERAGERARALGLGSTCQFQVANALDLPFASDSF
DWVWSLESGEHMPNKAQFLQEAWRVLKPGGRLILATWCHRPIDPGNGPLTADERRHLQA
IYDVYCLPYVVSLPDYEAIARECGFGEIKTADWSVAVAPFWDRVIESAFDPRVLWALGQ
AGPKIINAALCLRLMKWGYERGLVRFGLLTGIKPLV"
gene complement(9574..10527)
/gene="TMT"
/label="TMT"
/note="SyTMT"
RBS complement(10535..10557)
/label="RBS"
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
promoter complement(10692..10710)
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
This page is informational only.