LOCUS       Exported                7672 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Chloroplast transformation vector pTCH-RS2, complete sequence.
ACCESSION   KY426960
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7672)
  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 7672)
  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 7672)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7672
                     /organism="Chloroplast transformation vector pTCH-RS2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1982049"
     misc_feature    1..824
                     /note="plastid sequence targeting insertions in the 
                     trnI-trnA intergenic region II; similar to INSD accession 
                     Z00044"
     misc_feature    837..3000
                     /note="pUC118 vector sequence; similar to INSD accession 
                     U07649"
     rep_origin      complement(1073..1661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1832..2692)
                     /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(2693..2797)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3021..4438
                     /note="plastid sequence targeting insertions in the 
                     trnI-trnA intergenic region I; similar to INSD accession 
                     Z00044"
     regulatory      4470..4586
                     /regulatory_class="promoter"
                     /note="plastid rRNA operon PEP promoter; Prrn"
     regulatory      4591..4607
                     /regulatory_class="attenuator"
                     /note="psbA 5 UTR; similar to NC_001879"
     CDS             4610..5401
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside 3-adenylyltransferase"
                     /label=aadA
                     /note="confers the resistance to spectinomycin or 
                     streptomycin in bacteria and plants; similar to INSD 
                     accession AY442171"
                     /protein_id="ARK38551.1"
                     /translation="MAEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            4610..5401
                     /gene="aadA"
                     /label=aadA
     misc_recomb     5445..5478
                     /note="loxP recombination site; similar to INSD accession 
                     M10494"
     protein_bind    5445..5478
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_recomb     5517..5622
                     /note="ParA resolution site-MRS; similar to INSD accession 
                     M59825"
     misc_recomb     5634..5752
                     /note="CinH resolution site-RS2; similar to INSD accession 
                     AF213017"
     misc_recomb     5792..5843
                     /note="Bxb1 attP52 recognition site; similar to INSD 
                     accession JQ305140"
     misc_recomb     5904..5960
                     /note="phiC31 attP57 recognition site; similar to INSD 
                     accession GU56446"
     regulatory      6017..6166
                     /regulatory_class="terminator"
                     /note="rps16; rhct1 mRNA 3 maturation; similar to 
                     NC_001879"
     regulatory      6176..6278
                     /regulatory_class="promoter"
                     /note="psbA; similar to NC_001879"
     regulatory      6279..6363
                     /regulatory_class="attenuator"
                     /note="psbA 5 UTR; similar to NC_001879"
     CDS             6372..7055
                     /codon_start=1
                     /gene="DSRed"
                     /product="DSRed"
                     /label=DSRed
                     /note="red fluorescent protein from Discosoma sp.; similar 
                     to INSD accession AGE97310"
                     /protein_id="ARK38552.1"
                     /translation="MASSENVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRHH
                     LFLVP"
     gene            6372..7055
                     /gene="DSRed"
                     /label=DSRed
     CDS             6372..7046
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     misc_recomb     7062..7180
                     /note="similar to INSD accession JQ305140; spacer region 
                     has been modified to not match the original RS2; engineered
                     to inhibit homologous recombination in chloroplast; CinH 
                     resolution site recognition site - RS2*"
     regulatory      7205..7395
                     /regulatory_class="terminator"
                     /note="psbA 3 UTR; similar to NC_001879"
     misc_feature    7441..7619
                     /note="plastid DNA targeting sequence; similar to inverted 
                     repeat sequence from INSD accession Z00044"