LOCUS       Exported                3928 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector CMVp-HHRibo-gRNA1-HDVRibo-pA, complete sequence.
ACCESSION   KJ796498
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3928)
  AUTHORS   Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK.
  TITLE     Multiplexed and programmable regulation of gene networks with an 
            integrated RNA and CRISPR/Cas toolkit in human cells
  JOURNAL   Mol. Cell 54 (4), 698-710 (2014)
  PUBMED    24837679
REFERENCE   2  (bases 1 to 3928)
  AUTHORS   Perli SD.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2014) Electrical Engineering and Computer Science,
            Massachusetts Institute of Technology, 77, Massachusetts Avenue, 
            Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 3928)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3928
                     /organism="Cloning vector CMVp-HHRibo-gRNA1-HDVRibo-pA"
                     /mol_type="other DNA"
                     /db_xref="taxon:1520530"
     promoter        complement(620..724)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      751..1206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     regulatory      1335..1923
                     /regulatory_class="promoter"
                     /note="pmCMV"
     enhancer        1395..1698
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1699..1902
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    1930..1972
                     /label=5_prime HH self-cleaving ribozyme
                     /note="5_prime HH self-cleaving ribozyme"
     misc_feature    1973..2072
                     /label=gRNA1
                     /note="gRNA1"
     misc_RNA        1993..2068
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2073..2140
                     /label=3_prime HDV self-cleaving ribozyme
                     /note="3_prime HDV self-cleaving ribozyme"
     regulatory      2158..2607
                     /regulatory_class="polyA_signal_sequence"
                     /gene="HSV"
     gene            2158..2607
                     /gene="HSV"
                     /label=HSV
     polyA_signal    2389..2436
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      complement(2928..3516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3687..619)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="AIJ03097.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"