LOCUS       Exported                3192 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pm!sp-gg, complete sequence.
ACCESSION   MH015244
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3192)
  AUTHORS   Chavez A, Pruitt BW, Tuttle M, Shapiro RS, Cecchi RJ, Winston J, 
            Turczyk BM, Tung M, Collins JJ, Church GM.
  TITLE     Precise Cas9 targeting enables genomic mutation prevention
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. (2018) In press
  PUBMED    29555762
REFERENCE   2  (bases 1 to 3192)
  AUTHORS   Chavez A, Pruitt BW, Tuttle M, Shapiro RS, Cecchi RJ, Winston J, 
            Turczyk BM, Tung M, Collins JJ, Church GM.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAR-2018) Synthetic Biology, Wyss Institute, 3 
            Blackfan Circle, Floor 5, Boston, MA 02115, USA
REFERENCE   3  (bases 1 to 3192)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3192
                     /organism="Cloning vector pm!sp-gg"
                     /mol_type="other DNA"
                     /db_xref="taxon:2138192"
     regulatory      1..50
                     /regulatory_class="promoter"
                     /note="pLTetO promoter"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    26..44
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    67..87
                     /label=golden gate cassette
                     /note="golden gate cassette"
     misc_feature    complement(67..73)
                     /label=SapI
                     /note="SapI"
     misc_feature    74..80
                     /label=padding
                     /note="padding"
     misc_feature    81..87
                     /label=SapI
                     /note="SapI"
     misc_feature    88..123
                     /note="DR sequence; SP Cas9"
     repeat_region   88..123
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     CDS             complement(985..1518)
                     /codon_start=1
                     /gene="aac(3)I"
                     /product="Aac(3)I"
                     /label=aac(3)I
                     /protein_id="AVR59335.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(985..1518)
                     /gene="aac(3)I"
                     /label=aac(3)I
     promoter        complement(1707..1735)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      2164..3024
                     /label=p15A ori
                     /note="p15A ori"
     rep_origin      complement(2379..2924)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."