LOCUS       Exported               12759 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Chloroplast transformation vector pC35-STDBxb1, complete sequence.
ACCESSION   KF647530
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12759)
  AUTHORS   Shao M, Kumar S, Thomson JG.
  TITLE     Precise excision of plastid DNA by the large serine recombinase Bxb1
  JOURNAL   Plant Biotechnol. J. (2013) In press
  PUBMED    24261912
REFERENCE   2  (bases 1 to 12759)
  AUTHORS   Shao M, Kumar S, Thomson JG.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-SEP-2013) Crop Improvement and Utilization, USDA, 800 
            Buchanan Street, Albany, CA 94710, USA
REFERENCE   3  (bases 1 to 12759)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..12759
                     /organism="Chloroplast transformation vector pC35-STDBxb1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1429096"
     misc_feature    1..328
                     /label=pCambia 2300 vector sequence
                     /note="pCambia 2300 vector sequence"
     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 GenBank Accession 
                     Number V00140.1"
     promoter        1868..1914
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             1938..3620
                     /codon_start=1
                     /gene="STDBxb1"
                     /product="STDBxb1"
                     /label=STDBxb1
                     /note="Bxb1 large serine recombinase/integrase from 
                     Mycobacterium smegmati bacteriophage Bxb1; similar to INSD 
                     Accession Number JQ305141.1; stroma targeting domain (STD) 
                     fused to Bxb1 large serine recombinase"
                     /protein_id="AHB17669.1"
                     /translation="MASSLLSSAAVATRSNVAQANMVAPFTGLKSAASFPVSRKQNLDI
                     TSIASNGGRVQCMLINRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDL
                     DVSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKL
                     VVSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWG
                     YLPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQ
                     GREPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRA
                     ELVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGT
                     VAMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRA
                     GSPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRS
                     MNVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     gene            1938..3620
                     /gene="STDBxb1"
                     /label=STDBxb1
     sig_peptide     1938..2111
                     /gene="STDBxb1"
                     /note="tobacco gene for ribulose 1,5-bisphosphate 
                     carboxylase small subunit stroma targeting domain (STD); 
                     similar to INSD Accession Number X02353.1"
     regulatory      3626..4395
                     /regulatory_class="terminator"
                     /note="Nos3 terminator from Agrobacterium tumefaciens str.
                     C58 plasmid Ti; similar to INSD Accession Number 
                     AE007871.2"
     terminator      3795..4047
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    4396..12759
                     /label=pCambia 2300 vector sequence
                     /note="pCambia 2300 vector sequence"
     primer_bind     complement(4411..4427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4435..4451
                     /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(4459..4489)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4504..4525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4716..5393
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             5456..6253
                     /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    6310..6484
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    6562..6586
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             7011..7805
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      7892..8480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    8666..8806
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      9150..9344
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9410..10477)
                     /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(10911..11540)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"