Basic Vector Information
- Vector Name:
- pN-IC101
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9731 bp
- Type:
- Chloroplast transformation vector
- Replication origin:
- ori
- Source/Author:
- Lin CH, Chang ML, Tzeng CC, Chen LJ.
- Promoter:
- T3
pN-IC101 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pN-IC101 vector Sequence
LOCUS 40924_32958 9731 bp DNA circular SYN 18-DEC-2018
DEFINITION Chloroplast transformation vector pN-IC101, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9731)
AUTHORS Lin CH, Chang ML, Tzeng CC, Chen LJ.
TITLE Co-transformation of the synthetic cry1A(c) and wild-type cry1C
genes encoding Bacillus thuringiensis endotoxins into tobacco and
their bipartite expression effects on insecticidal efficacy
JOURNAL Zhi Wu Bao Hu Xue Hui Hui Kan 44, 209-232 (2002)
REFERENCE 2 (bases 1 to 9731)
AUTHORS Lin CH, Chen YY, Tzeng CC, Tsay HS, Chen LJ.
TITLE Expression of a Bacillus thuringiensis cry1C gene in plastid confers
high insecticidal efficacy against tobacco cutworm - a Spodoptera
insect
JOURNAL Bot. Bull. Acad. Sinica (Taiwan) 44, 199-210 (2003)
REFERENCE 3 (bases 1 to 9731)
AUTHORS Lin CH, Chen LJ.
TITLE Direct Submission
JOURNAL Submitted (17-OCT-2003) Institute of Molecular Biology, National
Chung Hsing University, 250, Kuokuang Rd., Taichung, Taiwan 402, ROC
REFERENCE 4 (bases 1 to 9731)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 9731)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Zhi Wu Bao
Hu Xue Hui Hui Kan 44, 209-232 (2002)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Bot. Bull.
Acad. Sinica (Taiwan) 44, 199-210 (2003)"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(17-OCT-2003) Institute of Molecular Biology, National Chung Hsing
University, 250, Kuokuang Rd., Taichung, Taiwan 402, ROC"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9731
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 121..137
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 144..162
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 172..1446
/gene="rbcL"
/label=rbcL
/note="Ribulose bisphosphate carboxylase large chain from
Buddleja davidii. Accession#: P36482"
3'UTR 1450..1719
/label=for rbcL mRNA 3' maturation
/note="for rbcL mRNA 3' maturation"
regulatory 1742..1859
/note="Prrn promoter in tobacco plastome; corresponding to
(102564..102681) or complementary(139945..140062) of the
tobacco plastome in accession number NC_001879; for
transcription of aadA gene"
/regulatory_class="promoter"
CDS 1877..2665
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
3'UTR 2679..3073
/note="psbA 3'-UTR; for aadA mRNA 3' maturation;
corresponds to nts 141-533 of tobacco plastome accession
number NC_001879"
5'UTR 3331..3378
/note="psbA-5'UTR; for translation initiation; derived from
nts 1596-1643 of tobacco plastome in accession number
NC_001879"
CDS 3379..5364
/codon_start=1
/gene="cry1C"
/product="Cry1C"
/label=cry1C
/note="toxic N terminal domain; insecticidal protein with
specificity to Spodoptera insects"
/protein_id="AAR14533.1"
/translation="MEENNQNQCIPYNCLSNPEEVLLDGERISTGNSSIDISLSLVQFL
VSNFVPGGGFLVGLIDFVWGIVGPSQWDAFLVQIEQLINERIAEFARNAAIANLEGLGN
NFNIYVEAFKEWEEDPNNPATRTRVIDRFRILDGLLERDIPSFRISGFEVPLLSVYAQA
ANLHLAILRDSVIFGERWGLTTINVNENYNRLIRHIDEYADHCANTYNRGLNNLPKSTY
QDWITYNRLRRDLTLTVLDIAAFFPNYDNRRYPIQPVGQLTREVYTDPLINFNPQLQSV
AQLPTFNVMESSAIRNPHLFDILNNLTIFTDWFSVGRNFYWGGHRVISSLIGGGNITSP
IYGREANQEPPRSFTFNGPVFRTLSNPTLRLLQQPWPAPPFNLRGVEGVEFSTPTNSFT
YRGRGTVDSLTELPPEDNSVPPREGYSHRLCHATFVQRSGTPFLTTGVVFSWTHRSATL
TNTIDPERINQIPLVKGFRVWGGTSVITGPGFTGGDILRRNTFGDFVSLQVNINSPITQ
RYRLRFRYASSRDARVIVLTGAASTGVGGQVSVNMPLQKTMEIGENLTSRTFRYTDFSN
PFSFRANPDIIGISEQPLFGAGSISSGELYIDKIEIILADATFEAESDLERAQKAVNAL
FTSSNQIGLKTDVTDYHIDQVSNLVE"
gene 3379..5364
/gene="cry1C"
/label=cry1C
primer_bind complement(5605..5621)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 5735..7025
/note="homologous recombination sequence; corresponds to
nts 59313-60603 of the tobacco plastome in accession number
NC_001879"
CDS 6215..7025
/codon_start=1
/gene="accD"
/product="AccD"
/label=accD
/note="ORF512"
/protein_id="AAR14534.1"
/translation="MTIHLLYFHANRGQENSMERWWFNSMLFKKEFERRCGLNKSMGSL
GPIENTNEDPNRKVKNIHSWRNRDNSSCSNVDYLFGVKDIRNFISDDTFLVSDRNGDSY
SIYFDIENHIFEIDNDHSFLSELESSFYSYRNSNYRNNGFRGEDPYYNSYMYDTQYSWN
NHINSCIDSYLQSQICIDTSIISGSENYGDSYIYRAVCGGESRNSSENEGSSRRTRTKG
SDLTIRESSNDLEVTQKYRHLWVQCENCYGLNYKKFLKSKMNICEQCG"
gene 6215..7025
/gene="accD"
/label=accD
promoter complement(7064..7082)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(7103..7119)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(7127..7143)
/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)."
promoter complement(7151..7181)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(7196..7217)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(7505..8093)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8267..9124)
/label=AmpR
/note="beta-lactamase"
promoter complement(9125..9229)
/label=AmpR promoter
rep_origin complement(9255..9710)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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