LOCUS       Exported                9731 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Chloroplast transformation vector pN-IC101, complete sequence.
ACCESSION   AY442171
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9731
                     /organism="Chloroplast transformation vector pN-IC101"
                     /mol_type="other DNA"
                     /db_xref="taxon:254404"
     misc_feature    join(1..185,7026..9731)
                     /label=pBluescript KS+ vector
                     /note="pBluescript KS+ vector"
     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"
     misc_feature    179..1719
                     /note="homologous recombination sequence; corresponds to 
                     nts 57758-59298 of the tobacco plastome in accession number
                     NC_001879"
     CDS             179..1449
                     /codon_start=1
                     /gene="rbcL"
                     /product="ribulose-1,5-bisphosphate carboxylase/oxygenase 
                     large subunit"
                     /label=rbcL
                     /protein_id="AAR14531.1"
                     /translation="PR*LPNLLLVHGQLYGPMDLPALIVTKGDATASSVLLEKKINILL
                     M*LTL*TFLKKVLLPTCLLPL*VTYLGSKPCALYVWKICESLLLMLKLSKVRLMGSKLK
                     EIN*TSMVVPCWDVLLNLNWGYLLKTTVEPFMNVFAVDLILLKMMRT*THNHLCVGEIV
                     SYFVPKHFIKHRLKQVKSKGIT*MLLQVHAKK*SKELYLLENWAFRS*CMTT*RGDSPQ
                     ILAWLIIAEIMVYFFTSTVQCMRLLIDRRIMVSTSGY*QKRYVCLVEIIFTLVP**VNL
                     KVKET*LWALLIYCVMILLNKIEVAVFISLKIGSLYQVFYPWLQEVFTFGICLL*PRSL
                     GMIPYYSSVEEL*DILGVMRQVP*LIE*L*KHV*KLVMKDVILLRKVMKLFARLANGAR
                     N*LLLVKYGKRSYLILQQWTFWIS"
     gene            179..1449
                     /gene="rbcL"
                     /label=rbcL
     3UTR           1450..1719
                     /label=for rbcL mRNA 3 maturation
                     /note="for rbcL mRNA 3 maturation"
     regulatory      1742..1859
                     /regulatory_class="promoter"
                     /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"
     CDS             1877..2668
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenine transferase"
                     /label=aadA
                     /note="selection marker; confers the resistance to 
                     spectinomycin or streptomycin in bacteria and plants"
                     /protein_id="AAR14532.1"
                     /translation="MAEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            1877..2668
                     /gene="aadA"
                     /label=aadA
     3UTR           2679..3073
                     /note="psbA 3-UTR; for aadA mRNA 3 maturation; 
                     corresponds to nts 141-533 of tobacco plastome accession 
                     number NC_001879"
     misc_feature    join(3080..3334,5372..5691)
                     /note="plastid transcription unit (pTZ19-PLsTa) comprising 
                     rbcL -180 promoter region from spinach chloroplast and 
                     the wild-type E. coli thra terminator"
     5UTR           3331..3378
                     /note="psbA-5UTR; 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    7127..7143
                     /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)."
     promoter        complement(7151..7181)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7196..7217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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(8264..9124)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9125..9229)
                     /gene="bla"
                     /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"