LOCUS       40924_10326        9690 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Non-integrating (episomal) expression of mouse p53DD - p53 
            carboxy-terminal dominant-negative fragment .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9690)
  AUTHORS   Okita K, Yamakawa T, Matsumura Y, Sato Y, Amano N, Watanabe A, 
            Goshima N, Yamanaka S
  TITLE     An Efficient Non-viral Method to Generate Integration-Free Human iPS
            Cells from Cord Blood and Peripheral Blood Cells.
  JOURNAL   Stem Cells. 2012 Nov 29. doi: 10.1002/stem.1293.
  PUBMED    23193063
REFERENCE   2  (bases 1 to 9690)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9690)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cells.
            2012 Nov 29. doi: 10.1002/stem.1293."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids,
                     forward primer"
     primer_bind     complement(2143..2162)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2208..2263
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2262..2281)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also
                     called rb-glob-pA-term-R"
     primer_bind     complement(2624..2640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2648..2664)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2672..2702)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2717..2738)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3217..5136
                     /codon_start=1
                     /product="Epstein-Barr nuclear antigen 1, also known as
                     EBNA-1"
                     /label=EBNA1
                     /note="crucial for latent viral infection, and for episomal
                     amplification of vectors containing the oriP origin (Okita 
                     et al., 2013)"
                     /translation="MSDEGPGTGPGNGLGEKGDTSGPEGSGGSGPQRRGGDNHGRGRGR
                     GRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGTHGGTGAGAGAGGAGAGGAG
                     AGGGAGAGGGAGGAGGAGGAGXGGGXGAGGGAGGXGGXGXGGGAGXGGGAGGAGAGGGA
                     GGGGGXGAGGGAGAGGGAGGAGAGGAGGAGGAGGGGGAGXGGGGGAGGAGAGGAGAGGG
                     AGGAGGAGAGGAGAGGAGAGGAGGGGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGG
                     AGGAGAGGGGGXGXGGXGGXGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRGGS
                     GGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPPPG
                     RRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGRRK
                     KGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSLYN
                     LRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLKDA
                     IKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDEGE
                     EGQE"
     rep_origin      5438..7227
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     primer_bind     complement(7768..7785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7939..8527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8701..9558)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9559..9663)
                     /label=AmpR promoter
     promoter        9690
                     /label=CAG
                     /note="CMV early enhancer fused to modified chicken 
                     beta-actin promoter"
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggsggggggg
      541 ggggssggbg kskcskrsrs gygcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgcccttca
     1741 ccaggatgac tgccatggag gagtcacagt cggatatcag cctcaagaga gcgctgccca
     1801 cctgcacaag cgcctctccc ccgcaaaaga aaaaaccact tgatggagag tatttcaccc
     1861 tcaagatccg cgggcgtaaa cgcttcgaga tgttccggga gctgaatgag gccttagagt
     1921 taaaggatgc ccatgctaca gaggagtctg gagacagcag ggctcactcc agctacctga
     1981 agaccaagaa gggccagtct acttcccgcc ataaaaaaac aatggtcaag aaagtggggc
     2041 ctgactcaga ctgaagggcg aattcactcc tcaggtgcag gctgcctatc agaaggtggt
     2101 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     2161 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     2221 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     2281 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     2341 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     2401 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     2461 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     2521 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     2581 atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg
     2641 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2701 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2761 ctttccagtc gggaaacctg tcgtgccagc ggatctcaat tctcatgttt gacagcttat
     2821 catcgataag ctgatcctca caggccgcac ccagcttttc ttccgttgcc ccagtagcat
     2881 ctctgtctgg tgaccttgaa gaggaagagg aggggtcccg agaatcccca tccctaccgt
     2941 ccagcaaaaa gggggacgag gaatttgagg cctggcttga ggctcaggac gcaaatcttg
     3001 aggatgttca gcgggagttt tccgggctgc gagtaattgg tgatgaggac gaggatggtt
     3061 cggaggatgg ggaattttca gacctggatc tgtctgacag cgaccatgaa ggggatgagg
     3121 gtgggggggc tgttggaggg ggcaggagtc tgcactccct gtattcactg agcgtcgtct
     3181 aataaagatg tctattgatc tcttttagtg tgaatcatgt ctgacgaggg gccaggtaca
     3241 ggacctggaa atggcctagg agagaaggga gacacatctg gaccagaagg ctccggcggc
     3301 agtggacctc aaagaagagg gggtgataac catggacgag gacggggaag aggacgagga
     3361 cgaggaggcg gaagaccagg agccccgggc ggctcaggat cagggccaag acatagagat
     3421 ggtgtccgga gaccccaaaa acgtccaagt tgcattggct gcaaagggac ccacggtgga
     3481 acaggagcag gagcaggagc gggaggggca ggagcaggag gggcaggagc aggaggaggg
     3541 gcaggagcag gaggaggggc aggaggggca ggaggggcag gaggggcagg agsaggagga
     3601 ggggsaggag caggaggagg ggcaggaggg gsaggagggg saggagsagg aggaggggca
     3661 ggagsaggag gaggggcagg aggggcagga gcaggaggag gggcaggagg gggaggaggg
     3721 gsaggagcag gaggaggggc aggagcagga ggaggggcag gaggggcagg agcaggaggg
     3781 gcaggagggg caggaggggc aggaggagga ggaggggcag gagsaggagg gggaggaggg
     3841 gcaggagggg caggagcagg aggggcagga gcaggaggag gggcaggagg ggcaggaggg
     3901 gcaggagcag gaggggcagg agcaggaggg gcaggagcag gaggggcagg aggrggaggr
     3961 gcaggagggg caggagcagg aggggcagga ggggcaggag caggaggggc aggaggggca
     4021 ggagcaggag gggcaggagg ggcaggagca ggaggagggg caggaggggc aggagcagga
     4081 gggggaggag gggsaggags aggaggggsa ggaggggsag gagcaggagg ggcaggaggg
     4141 gcaggagcag gaggaggggc aggagcagga ggggcaggag caggaggtgg aggccggggt
     4201 cgaggaggca gtggaggccg gggtcgagga ggtagtggag gccggggtcg aggaggtagt
     4261 ggaggccgcc ggggtagagg acgtgaaaga gccagggggg gaagtcgtga aagagccagg
     4321 gggagaggtc gtggacgtgg agaaaagagg cccaggagtc ccagtagtca gtcatcatca
     4381 tccgggtctc caccgcgcag gccccctcca ggtagaaggc catttttcca ccctgtaggg
     4441 gaagccgatt attttgaata ccaccaagaa ggtggcccag atggtgagcc tgacgtgccc
     4501 ccgggagcga tagagcaggg ccccgcagat gacccaggag aaggcccaag cactggaccc
     4561 cggggtcagg gtgatggagg caggcgcaaa aaaggagggt ggtttggaaa gcatcgtggt
     4621 caaggaggtt ccaacccgaa atttgagaac attgcagaag gtttaagagc tctcctggct
     4681 aggagtcacg tagaaaggac taccgacgaa ggaacttggg tcgccggtgt gttcgtatat
     4741 ggaggtagta agacctccct ttacaaccta aggcgaggaa ctgcccttgc tattccacaa
     4801 tgtcgtctta caccattgag tcgtctcccc tttggaatgg cccctggacc cggcccacaa
     4861 cctggcccgc taagggagtc cattgtctgt tatttcatgg tctttttaca aactcatata
     4921 tttgctgagg ttttgaagga tgcgattaag gaccttgtta tgacaaagcc cgctcctacc
     4981 tgcaatatca gggtgactgt gtgcagcttt gacgatggag tagatttgcc tccctggttt
     5041 ccacctatgg tggaaggggc tgccgcggag ggtgatgacg gagatgacgg agatgaagga
     5101 ggtgatggag atgagggtga ggaagggcag gagtgatgta acttgttagg agacgccctc
     5161 aatcgtatta aaagccgtgt attcccccgc actaaagaat aaatccccag tagacatcat
     5221 gcgtgctgtt ggtgtatttc tggccatctg tcttgtcacc attttcgtcc tcccaacatg
     5281 gggcaattgg gcatacccat gttgtcacgt cactcagctc cgcgctcaac accttctcgc
     5341 gttggaaaac attagcgaca tttacctggt gagcaatcag acatgcgacg gctttagcct
     5401 ggcctcctta aattcaccta agaatgggag caaccagcag gaaaaggaca agcagcgaaa
     5461 attcacgccc ccttgggagg tggcggcata tgcaaaggat agcactccca ctctactact
     5521 gggtatcata tgctgactgt atatgcatga ggatagcata tgctacccgg atacagatta
     5581 ggatagcata tactacccag atatagatta ggatagcata tgctacccag atatagatta
     5641 ggatagccta tgctacccag atataaatta ggatagcata tactacccag atatagatta
     5701 ggatagcata tgctacccag atatagatta ggatagccta tgctacccag atatagatta
     5761 ggatagcata tgctacccag atatagatta ggatagcata tgctatccag atatttgggt
     5821 agtatatgct acccagatat aaattaggat agcatatact accctaatct ctattaggat
     5881 agcatatgct acccggatac agattaggat agcatatact acccagatat agattaggat
     5941 agcatatgct acccagatat agattaggat agcctatgct acccagatat aaattaggat
     6001 agcatatact acccagatat agattaggat agcatatgct acccagatat agattaggat
     6061 agcctatgct acccagatat agattaggat agcatatgct atccagatat ttgggtagta
     6121 tatgctaccc atggcaacat tagcccaccg tgctctcagc gacctcgtga atatgaggac
     6181 caacaaccct gtgcttggcg ctcaggcgca agtgtgtgta atttgtcctc cagatcgcag
     6241 caatcgcgcc cctatcttgg cccgcccacc tacttatgca ggtattcccc ggggtgccat
     6301 tagtggtttt gtgggcaagt ggtttgaccg cagtggttag cggggttaca atcagccaag
     6361 ttattacacc cttattttac agtccaaaac cgcagggcgg cgtgtggggg ctgacgcgtg
     6421 cccccactcc acaatttcaa aaaaaagagt ggccacttgt ctttgtttat gggccccatt
     6481 ggcgtggagc cccgtttaat tttcgggggt gttagagaca accagtggag tccgctgctg
     6541 tcggcgtcca ctctctttcc ccttgttaca aatagagtgt aacaacatgg ttcacctgtc
     6601 ttggtccctg cctgggacac atcttaataa ccccagtatc atattgcact aggattatgt
     6661 gttgcccata gccataaatt cgtgtgagat ggacatccag tctttacggc ttgtccccac
     6721 cccatggatt tctattgtta aagatattca gaatgtttca ttcctacact agtatttatt
     6781 gcccaagggg tttgtgaggg ttatattggt gtcatagcac aatgccacca ctgaaccccc
     6841 cgtccaaatt ttattctggg ggcgtcacct gaaaccttgt tttcgagcac ctcacataca
     6901 ccttactgtt cacaactcag cagttattct attagctaaa cgaaggagaa tgaagaagca
     6961 ggcgaagatt caggagagtt cactgcccgc tccttgatct tcagccactg cccttgtgac
     7021 taaaatggtt cactaccctc gtggaatcct gaccccatgt aaataaaacc gtgacagctc
     7081 atggggtggg agatatcgct gttccttagg acccttttac taaccctaat tcgatagcat
     7141 atgcttcccg ttgggtaaca tatgctattg aattagggtt agtctggata gtatatacta
     7201 ctacccggga agcatatgct acccgtttag ggttaacaag ggggccttat aaacactatt
     7261 gctaatgccc tcttgagggt ccgcttatcg gtagctacac aggcccctct gattgacgtt
     7321 ggtgtagcct cccgtagtct tcctgggccc ctgggaggta catgtccccc agcattggtg
     7381 taagagcttc agccaagagt tacacataaa ggcaatgttg tgttgcagtc cacagactgc
     7441 aaagtctgct ccaggatgaa agccactcag tgttggcaaa tgtgcacatc catttataag
     7501 gatgtcaact acagtcagag aacccctttg tgtttggtcc ccccccgtgt cacatgtgga
     7561 acagggccca gttggcaagt tgtaccaacc aactgaaggg attacatgca ctgccccgcg
     7621 aagaaggggc agagatgtcg tagtcaggtt tagttcgtcc ggggcgggga tcgatcctct
     7681 agagtcgacg ctagcggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7741 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7801 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7861 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7921 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7981 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8041 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8101 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     8161 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8221 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8281 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8341 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     8401 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8461 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8521 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8581 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8641 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8701 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8761 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8821 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8881 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8941 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     9001 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     9061 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     9121 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     9181 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9241 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9301 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9361 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9421 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9481 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9541 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9601 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9661 gcgcacattt ccccgaaaag tgccacctgg
//
