LOCUS       sequence-80596-e        1040 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Doxycycline-inducible lentiviral expression of SpCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1040)
  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 1040)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.1246981"; journalName: "Science"; date: "2014-01-3-
            3"; volume: "343"; issue: "6166"; pages: "80-4"
COMMENT     Sequence Label: pCW-Cas9
FEATURES             Location/Qualifiers
     source          1..1040
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(52..60,61..96,97..99,100..135,136..138,
                     139..174,175..180,181..798))
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             complement(799..852)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(933..953)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 aaaggagcaa catagttaag aatacgatat cttggatcga tccttactta gttacccggg
       61 gagcatgtca aggtcaaaat cgtcaagagc gtcagcaggc agcatatcaa ggtcaaagtc
      121 gtcaagggca tcggctggga gcatgtctaa gtcaaaatcg tcaagggcgt cggtcggccc
      181 gccgctttcg cactttagct gtttctccag gccacatatg attagttcca ggccgaaaag
      241 gaaggcaggt tcggctccct gccggtcgaa cagctcaatt gcttgtctca gaagtggggg
      301 catagaatcg gtggtaggtg tctctctttc ctcttttgct acttgatgct cctgttcctc
      361 caatacgcag cccagtgtaa agtggcccac ggcggacaga gcgtacagtg cgttctccag
      421 ggagaagcct tgctgacaca ggaacgcgag ctgattttcc agggtttcgt actgtttctc
      481 tgttgggcgg gtgccgagat gcactttagc cccgtcgcga tgtgagagga gagcacagcg
      541 gtatgacttg gcgttgttcc gcagaaagtc ttgccatgac tcgccttcca gggggcagaa
      601 gtgggtatga tgcctgtcca gcatctcgat tggcagggca tcgagcaggg cccgcttgtt
      661 cttcacgtgc cagtacaggg taggctgctc aactcccagc ttttgagcga gtttccttgt
      721 cgtcaggcct tcgataccga caccattgag taattccaga gctccgttta tgactttgct
      781 cttgtccagt ctagacattg gaccagggtt ttcttcaaca tcaccacaag tgaggagaga
      841 acctctacct tcggcaccgg gcttgcgggt catgcaccag gtgcgcggtc cttcgggcac
      901 ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt tgcggggcgc
      961 ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc cggggagcac
     1021 gacggcgctg cccagaccct
//
