LOCUS       Exported                1146 bp ds-DNA     linear   SYN 18-MAY-2021
DEFINITION  Doxycycline-inducible lentiviral expression of SpCas9.
ACCESSION   .
VERSION     .
KEYWORDS    pCW-Cas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1146)
  AUTHORS   Wang T, Wei JJ, Sabatini DM, Lander ES
  TITLE     Genetic screens in human cells using the CRISPR-Cas9 system.
  JOURNAL   Science. 2014 Jan 3;343(6166):80-4. doi: 10.1126/science.1246981. 
            Epub 2013 Dec 12.
  PUBMED    24336569
REFERENCE   2  (bases 1 to 1146)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 18, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..1146
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(330..350)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(357..422)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      422..431
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(448..762)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     primer_bind     complement(451..475)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    522..540
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    558..576
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    593..611
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    629..647
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    665..683
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    700..718
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    736..754
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    816..933
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tcttccagtc tgtggaagaa gctgtcgtcc accttggcca tctcgttgct gaagatctct
       61 tgcagatagc agatccggtt cttccgtctg gtgtatcttc ttctggcggt tctcttcagc
      121 cgggtggcct cggctgtttc gccgctgtcg aacagcaggg ctccgatcag gttcttcttg
      181 atgctgtgcc ggtcggtgtt gcccagcacc ttgaatttct tgctgggcac cttgtactcg
      241 tcggtgatca cggcccagcc cacagagttg gtgccgatgt ccaggccgat gctgtacttc
      301 ttgtcggctg ctgggactcc gtggataccg accttccgct tcttctttgg ggccatctta
      361 tcgtcatcgt ctttgtaatc aatatcatga tccttgtagt ctccgtcgtg gtccttatag
      421 tccatggtgg cgctagccaa ttctccaggc gatctgacgg ttcactaaac gagctctgct
      481 tatataggcc tcccaccgta cacgcctacc tcgacatacg ttctctatca ctgataggga
      541 gtaaactcga catacgttct ctatcactga tagggataaa ctcgacatac gttctctatc
      601 actgataggg agtaaactcg acatacgttc tctatcactg atagggagta aactcgacat
      661 acgttctcta tcactgatag ggagtaaact cgacatcgtt ctctatcact gatagggagt
      721 aaactcgaca tacgttctct atcactgata gggagtaaac tcgacatatc gattcgcggc
      781 caaagtggat ctctgctgtc cctgtaataa acccgaaaat tttgaatttt tgtaatttgt
      841 ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct ttcccctgca
      901 ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc catttgtctc
      961 gaggtcgaga attgtcccct cggggttggg aggtgggtct gaaacgataa tggtgaatat
     1021 ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa acctaccaag
     1081 cctcctacta tcattatgaa taattttata taccacagcc aatttgttat gttaaaccaa
     1141 ttccac
//
