LOCUS       Exported               11823 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pC35-STDCinHwt, complete sequence.
ACCESSION   KY426962
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11823)
  AUTHORS   Shao M, Blechl A, Thomson JG.
  TITLE     Small Serine Recombination Systems ParA-MRS and CinH-RS2 Perform 
            Precise Excision of Plastid DNA
  JOURNAL   Plant Biotechnol. J. (2017) In press
  PUBMED    28421718
REFERENCE   2  (bases 1 to 11823)
  AUTHORS   Shao M, Blechl A, Thomson JG.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JAN-2017) Crop Improvement and Utilization, USDA, 800 
            Buchanan Street, Albany, CA 94710, USA
REFERENCE   3  (bases 1 to 11823)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..11823
                     /organism="Expression vector pC35-STDCinHwt"
                     /mol_type="other DNA"
                     /db_xref="taxon:1982050"
     misc_feature    1..328
                     /note="pCambia2300 backbone; similar to INSD accession 
                     AF234315"
     misc_feature    82..106
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     309..325
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      329..1922
                     /regulatory_class="promoter"
                     /note="CaMV 35S promoter; similar to INSD accession V00140"
     promoter        1868..1914
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     regulatory      1933..1942
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1939..2685
                     /codon_start=1
                     /gene="STDCinH"
                     /product="STDCinH"
                     /label=STDCinH
                     /note="site-specific recombinase CinH; originally isolated 
                     from Acinetobacter sp. plasmid pKLH208B in INSD accession 
                     AJ251272; modified to include tobacco gene for ribulose 
                     1,5-bisphosphate carboxylase small subunit stroma targeting
                     domain (STD) from INSD accession X02353"
                     /protein_id="ARK38554.1"
                     /translation="MASSLLSSAAVATRSNVAQANMVAPFTGLKSAASFPVSRKQNLDI
                     TSIASNGGRVQCMLINKGQKVGYVRVSSVEQNTGRQLEGIEVDRIFVDRASGKNTDRPK
                     FQEMLNYVREGDRVIVHSMDRFARSLKDLVTEVDKLVKRGIAIQFVKENITFTAQSTPM
                     DNLMLQLMGAFAQFEREIILERQKEGIKIAAAQGKYKGRVHKLNPDQAKALLQAWKEGK
                     YSSKVDLAKAFGISRQAVYRYLKQIN"
     gene            1939..2685
                     /gene="STDCinH"
                     /label=STDCinH
     sig_peptide     1939..2112
                     /gene="STDCinH"
                     /note="stroma targeting domain; STD"
     regulatory      2860..3112
                     /regulatory_class="terminator"
                     /note="Nos3 terminator from Agrobacterium tumefaciens str.
                     C58 plasmid Ti in INSD accession AE007871"
     terminator      2860..3112
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3113..11823
                     /note="pCambia2300 backbone; similar to INSD accession 
                     AF234315"
     primer_bind     complement(3476..3492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3500..3516
                     /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(3524..3554)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3569..3590
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3781..4458
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             4521..5318
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5375..5549
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    5627..5651
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6076..6870
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      6957..7545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    7731..7871
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      8215..8409
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8475..9542)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     CDS             complement(9976..10605)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"