pEN-2xChimera vector (V007566)

LOCUS       40924_17399        4236 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEN-2xChimera, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4236)
  AUTHORS   Durr J, Papareddy R, Nakajima K, Gutierrez-Marcos J.
  TITLE     Highly efficient heritable targeted deletions of gene clusters and 
            non-coding regulatory regions in Arabidopsis using CRISPR/Cas9
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4236)
  AUTHORS   Durr J, Gutierrez-Marcos J.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-JAN-2017) School of Life Sciences, University of 
            Warwick, Gibbet Hill Campus, Coventry, West Midlands CV4 7AL, United
            Kingdom
REFERENCE   3  (bases 1 to 4236)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4236)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-JAN-2017) School of Life Sciences, University of Warwick, Gibbet
            Hill Campus, Coventry, West Midlands CV4 7AL, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      33..42
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      37..46
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    52..151
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      152..539
                     /label=U6-26
                     /note="U6-26"
                     /regulatory_class="promoter"
     misc_RNA        558..633
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(645..669)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      681..1068
                     /label=U6-26
                     /note="U6-26"
                     /regulatory_class="promoter"
     misc_feature    1085..1201
                     /label=sgRNA scaffold
                     /note="sgRNA scaffold"
     misc_RNA        1085..1160
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(1172..1196)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1226..1325)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1361..1379)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1397..1413)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1421..1437)
                     /label=lac operator
                     /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(1445..1475)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1490..1511)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1799..2387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2561..3418)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3419..3523)
                     /label=AmpR promoter
     rep_origin      complement(3601..4056)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     4197..4213
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4220..4236
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"