LOCUS       Exported                5010 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMono_T-vector, complete sequence.
ACCESSION   JN681269
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5010)
  AUTHORS   Fursova O, Pogorelko G, Zabotina OA.
  TITLE     An efficient method for transient gene expression in monocots 
            applied to modify the Brachypodium distachyon cell wall
  JOURNAL   Ann. Bot. 110 (1), 47-56 (2012)
  PUBMED    22589326
REFERENCE   2  (bases 1 to 5010)
  AUTHORS   Pogorelko GV, Fursova OV, Zabotina OA.
  TITLE     Entry T-vector with monocothyledonous plants specific elements 
            (mono_T-Vector)
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5010)
  AUTHORS   Pogorelko GV, Fursova OV, Zabotina OA.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-SEP-2011) BBMB, Iowa State University, ISU, Ames, IA 
            50011, USA
REFERENCE   4  (bases 1 to 5010)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5010
                     /organism="Cloning vector pMono_T-vector"
                     /mol_type="other DNA"
                     /db_xref="taxon:1133977"
     regulatory      33..42
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      37..46
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    61..162
                     /label=Zea mays ubiquitin signal peptide sequence
                     /note="Zea mays ubiquitin signal peptide sequence"
     misc_feature    179..1025
                     /note="fragment for XcmI restriction enzyme removal 
                     providing single T-overhand ends for TA direct cloning of 
                     PCR fragments modified by removal of first A from start 
                     ATG-codon and stop codon to fit into the same ORF with 
                     signal peptide and GFP"
     CDS             complement(189..1004)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AEX93341.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1064..1780
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=green fluorescent protein
                     /note="GFP; plant optimized"
                     /protein_id="AEX93343.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      1800..2046
                     /regulatory_class="terminator"
                     /map="NOS terminator"
     regulatory      complement(2104..2207)
                     /regulatory_class="promoter"
                     /note="Sp6"
     promoter        complement(2135..2153)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2171..2187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2195..2211
                     /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(2219..2249)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2264..2285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2378..3271
                     /label=ColE1 replication origin
                     /note="ColE1 replication origin"
     rep_origin      complement(2573..3161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3332..4192)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin resistance"
                     /protein_id="AEX93342.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4193..4297)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4368..4830)
                     /direction=LEFT
                     /label=F1 origin
                     /note="F1 origin"
     rep_origin      complement(4375..4830)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     4971..4987
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4994..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"