LOCUS       Exported                5386 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gene silencing vector pJRTCLCrVA.008, complete sequence.
ACCESSION   EU541443
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5386)
  AUTHORS   Tuttle JR, Idris AM, Brown JK, Haigler CH, Robertson D.
  TITLE     Geminivirus-mediated gene silencing from Cotton leaf crumple virus 
            is enhanced by low temperature in cotton
  JOURNAL   Plant Physiol. 148 (1), 41-50 (2008)
  PUBMED    18621976
REFERENCE   2  (bases 1 to 5386)
  AUTHORS   Tuttle JR III, Robertson D.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-FEB-2008) Plant Biology, North Carolina State 
            University, Box 7612 Gardner Hall, Raleigh, NC 27695, USA
REFERENCE   3  (bases 1 to 5386)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5386
                     /organism="Gene silencing vector pJRTCLCrVA.008"
                     /lab_host="Gossypium hirsutum"
                     /mol_type="other DNA"
                     /note="Geminivirus-based silencing vector; coat protein 
                     replaced with polylinker in A component of Cotton leaf 
                     crumple virus"
                     /db_xref="taxon:512085"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="confers ampicillin/carbinicillin resistance"
                     /protein_id="ACB32236.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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)."
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    2625..2672
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             complement(2727..3125)
                     /codon_start=1
                     /product="AL3"
                     /label=AL3
                     /note="replication enhancer; Ren"
                     /protein_id="ACB32239.1"
                     /translation="MDSRTGEIITARQAENGVYIWEIINPLYFKIYGVEDLLYTRTRIY
                     HVQIRFNHNLRRALHLHKAYLNFQVWTTSLTASGSTYLIRFKQLVMLYLDELGVISLNN
                     VIRAVRFATDKSYVSYVLENHSIKFKFY"
     CDS             complement(2872..3291)
                     /codon_start=1
                     /product="AL2"
                     /label=AL2
                     /note="transactivator of coat protein and BR1; Trap"
                     /protein_id="ACB32238.1"
                     /translation="MRRSNIGLSRMLSSSLSTPPSIKKAHRQAKRRAIRRRRIDLNCGC
                     TIYFHIGCTGHGFTHRGNHHCTSGREWRVYLGDNKSPIFQDIRSRGSAIHQDAYIPRPD
                     PIQPQLTESVASPQSLPELPSLDDISDSFWVDLFN"
     CDS             complement(3188..4252)
                     /codon_start=1
                     /product="AL1"
                     /label=AL1
                     /note="replication initiator; Rep"
                     /protein_id="ACB32237.1"
                     /translation="MPRNPDSFRLQARHIFLTYPKCDIPKNEALQMLQTLSWSVVKPTY
                     IRVAREEHSDGFPHLHCLIQLSGKSNIKNKRFFDLTHPRRSTNFHPNIQAAKDANAVKN
                     YITKDGDCCESGQFKVPGGTKKNKDDVYYNAINAPSLAEALAIIRAGDPRAFIVSYHNI
                     TANLERLFKKAPEQWVPPFPLSSFTNVPEQMQEWADDYFGMGPAARPVRPVSLIVEGDS
                     RTGKTMWARALGPHNYLSGHLDFNNRVYSNNVAYNVIDDVAPQYLKLKHWKELLGAQKD
                     WQSNCKYGKPVQIKGGIPSIVLCNPGEGSSYKDFLDKAENASLKHWTLKNVIFITLDAP
                     LYQEGTQASQEEGD"
     CDS             complement(3778..4173)
                     /codon_start=1
                     /product="AL4"
                     /label=AL4
                     /protein_id="ACB32240.1"
                     /translation="MKLFKCFKPYHGQSSNQHIFELPERSTRTDSHTFTVSSNSLESPT
                     SRIRDFSTLLTPEGRPIFTQTYKQPKTPTPSRITSPKTAIVVNPDSSKCLGVQRRIKTT
                     SIITPSMHLVWRKLLQLSGPEIQGRSS"
     promoter        complement(4743..4761)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4771..4787)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4929..5384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"