pN-IC101 vector (V004370)

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 Map

pN-IC1019731 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600M13 fwdT7 promoterRibulose bisphosphate carboxylase large chain3'UTRPrrn 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 geneSmR3'UTR5'UTRCry1CM13 fwdhomologous recombination sequence; corresponds to nts 59313-60603 of the tobacco plastome in accession number NC_001879T3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterf1 ori

pN-IC101 vector Sequence

LOCUS       V004370                 9731 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004370
VERSION     V004370
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)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Bot. Bull.
            Acad. Sinica (Taiwan) 44, 199-210 (2003)"
            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"
            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="Ribulose bisphosphate carboxylase large chain"
                     /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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