LOCUS       Exported                5550 bp ds-DNA     linear   SYN 12-JUN-2021
DEFINITION  EBNA episome plasmid for CAG promoter constitutive expression of 
            dCas9-VP192-P300core followed by T2A-GFP as a reporter. Includes p53
            shRNA expression cassette..
ACCESSION   .
VERSION     .
KEYWORDS    pCXLE-dCas9VPP300-T2A-GFP-shP53
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5550)
  AUTHORS   Weltner J, Balboa D, Katayama S, Bespalov M, Krjutskov K, Jouhilahti
            EM, Trokovic R, Kere J, Otonkoski T
  TITLE     Human pluripotent reprogramming with CRISPR activators.
  JOURNAL   Nat Commun. 2018 Jul 6;9(1):2643. doi: 10.1038/s41467-018-05067-x.
  PUBMED    29980666
REFERENCE   2  (bases 1 to 5550)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5550
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    complement(168..201)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      388..2177
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     promoter        2647..2960
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2879..2899
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     complement(3111..3128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3282..3870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3362..3381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4041..4901)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4664..4683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4902..5006)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5037..5416
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgtaacttgt taggagacgc cctcaatcgt attaaaagcc gtgtattccc ccgcactaaa
       61 gaataaatcc ccagtagaca tcatgcgtgc tgttggtgta tttctggcca tctgtcttgt
      121 caccattttc gtcctcccaa catggggcaa ttgccggaac ccttaatata acttcgtata
      181 atgtatgcta tacgaagtta ttaggtccct cgaagaggtt cactagcgga tctcaattgg
      241 gcatacccat gttgtcacgt cactcagctc cgcgctcaac accttctcgc gttggaaaac
      301 attagcgaca tttacctggt gagcaatcag acatgcgacg gctttagcct ggcctcctta
      361 aattcaccta agaatgggag caaccagcag gaaaaggaca agcagcgaaa attcacgccc
      421 ccttgggagg tggcggcata tgcaaaggat agcactccca ctctactact gggtatcata
      481 tgctgactgt atatgcatga ggatagcata tgctacccgg atacagatta ggatagcata
      541 tactacccag atatagatta ggatagcata tgctacccag atatagatta ggatagccta
      601 tgctacccag atataaatta ggatagcata tactacccag atatagatta ggatagcata
      661 tgctacccag atatagatta ggatagccta tgctacccag atatagatta ggatagcata
      721 tgctacccag atatagatta ggatagcata tgctatccag atatttgggt agtatatgct
      781 acccagatat aaattaggat agcatatact accctaatct ctattaggat agcatatgct
      841 acccggatac agattaggat agcatatact acccagatat agattaggat agcatatgct
      901 acccagatat agattaggat agcctatgct acccagatat aaattaggat agcatatact
      961 acccagatat agattaggat agcatatgct acccagatat agattaggat agcctatgct
     1021 acccagatat agattaggat agcatatgct atccagatat ttgggtagta tatgctaccc
     1081 atggcaacat tagcccaccg tgctctcagc gacctcgtga atatgaggac caacaaccct
     1141 gtgcttggcg ctcaggcgca agtgtgtgta atttgtcctc cagatcgcag caatcgcgcc
     1201 cctatcttgg cccgcccacc tacttatgca ggtattcccc ggggtgccat tagtggtttt
     1261 gtgggcaagt ggtttgaccg cagtggttag cggggttaca atcagccaag ttattacacc
     1321 cttattttac agtccaaaac cgcagggcgg cgtgtggggg ctgacgcgtg cccccactcc
     1381 acaatttcaa aaaaaagagt ggccacttgt ctttgtttat gggccccatt ggcgtggagc
     1441 cccgtttaat tttcgggggt gttagagaca accagtggag tccgctgctg tcggcgtcca
     1501 ctctctttcc ccttgttaca aatagagtgt aacaacatgg ttcacctgtc ttggtccctg
     1561 cctgggacac atcttaataa ccccagtatc atattgcact aggattatgt gttgcccata
     1621 gccataaatt cgtgtgagat ggacatccag tctttacggc ttgtccccac cccatggatt
     1681 tctattgtta aagatattca gaatgtttca ttcctacact agtatttatt gcccaagggg
     1741 tttgtgaggg ttatattggt gtcatagcac aatgccacca ctgaaccccc cgtccaaatt
     1801 ttattctggg ggcgtcacct gaaaccttgt tttcgagcac ctcacataca ccttactgtt
     1861 cacaactcag cagttattct attagctaaa cgaaggagaa tgaagaagca ggcgaagatt
     1921 caggagagtt cactgcccgc tccttgatct tcagccactg cccttgtgac taaaatggtt
     1981 cactaccctc gtggaatcct gaccccatgt aaataaaacc gtgacagctc atggggtggg
     2041 agatatcgct gttccttagg acccttttac taaccctaat tcgatagcat atgcttcccg
     2101 ttgggtaaca tatgctattg aattagggtt agtctggata gtatatacta ctacccggga
     2161 agcatatgct acccgtttag ggttaacaag ggggccttat aaacactatt gctaatgccc
     2221 tcttgagggt ccgcttatcg gtagctacac aggcccctct gattgacgtt ggtgtagcct
     2281 cccgtagtct tcctgggccc ctgggaggta catgtccccc agcattggtg taagagcttc
     2341 agccaagagt tacacataaa ggcaatgttg tgttgcagtc cacagactgc aaagtctgct
     2401 ccaggatgaa agccactcag tgttggcaaa tgtgcacatc catttataag gatgtcaact
     2461 acagtcagag aacccctttg tgtttggtcc ccccccgtgt cacatgtgga acagggccca
     2521 gttggcaagt tgtaccaacc aactgaaggg attacatgca ctgccccgcg aagaaggggc
     2581 agagatgtcg tagtcaggtt tagttcgtcc ggggcgggga tcgatcctct agagtcgacg
     2641 ctagcggatc cgacgccgcc atctctaggc ccgcgccggc cccctcgcac agacttgtgg
     2701 gagaagctcg gctactcccc tgccccggtt aatttgcata taatatttcc tagtaactat
     2761 agaggcttaa tgtgcgataa aagacagata atctgttctt tttaatacta gctacatttt
     2821 acatgatagg cttggatttc tataagagat acaaatacta aattattatt ttaaaaaaca
     2881 gcacaaaagg aaactcaccc taactgtaaa gtaattgtgt gttttgagac tataaatatc
     2941 ccttggagaa aagccttgtt tgggccgact ccagtggtaa tctacttcaa gagagtagat
     3001 taccactgga gtcttttttg gaattcctgc agcccggggg atccgctgca ttaatgaatc
     3061 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3121 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3181 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3241 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3301 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3361 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3421 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3481 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3541 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3601 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3661 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3721 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3781 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3841 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3901 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3961 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4021 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4081 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4141 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4201 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4261 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4321 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4381 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4441 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4501 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4561 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4621 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4681 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4741 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4801 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4861 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4921 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4981 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca
     5041 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     5101 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5161 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5221 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5281 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     5341 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     5401 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     5461 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
     5521 ggcggggggg gggggggggc gcgcgccagg
//
