LOCUS       40924_13930        8495 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Non-integrating (episomal) expression of mouse Oct3/4, Klf4 and 
            Sox2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8495)
  AUTHORS   Okita K, Nakagawa M, Hyenjong H, Ichisaka T, Yamanaka S
  TITLE     Generation of Mouse Induced Pluripotent Stem Cells Without Viral 
            Vectors.
  JOURNAL   Science. 2008 Oct 9. ():.
  PUBMED    18845712
REFERENCE   2  (bases 1 to 8495)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8495)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2008 Oct 9. ():."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8495
                     /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"
     CDS             1762..2817
                     /codon_start=1
                     /label=mPou5f1
                     /note="Mus musculus Oct-4 gene. Encodes a transcription
                     factor containing a POU homeodomain that plays a key role 
                     in embryonic development and stem cell pluripotency."
                     /translation="MAGHLASDFAFSPPPGGGDGSAGLEPGWVDSRTWLSFQGPPGGPG
                     IGPGSEVLGISPCPPAYEFCGGMAYCGPQVGLGLVPQVGVETLQPEGQAGARVESNSEG
                     TSSEPCADRPNAVKLEKVEPTPEESQDMKALQKELEQFAKLLKQKRITLGYTQADVGLT
                     LGVLFGKVFSQTTICRFEALQLSLKNMCKLRPLLEKWVEEADNNENLQEICKSETLVQA
                     RKRKRTSIENRVRWSLETMFLKCPKPSLQQITHIANQLGLEKDVVRVWFCNRRQKGKRS
                     SIEYSQREEYEATGTPFPGGAVSFPLPPGPHFGTPGYGSPHFTTLYSVPFPEGEAFPSV
                     PVTALGSPMHSN"
     CDS             2842..2907
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             2911..4332
                     /codon_start=1
                     /label=mKlf4
                     /note="Mouse Kruppel-like factor (Klf4) gene. Belongs to
                     the relatively large family of SP1-like transcription 
                     factors and is involved in the regulation of proliferation,
                     differentiation, apoptosis and somatic cell reprogramming."
                     /translation="MAVSDALLPSFSTFASGPAGREKTLRPAGAPTNRWREELSHMKRL
                     PPLPGRPYDLAATVATDLESGGAGAACSSNNPALLARRETEEFNDLLDLDFILSNSLTH
                     QESVAATVTTSASASSSSSPASSGPASAPSTCSFSYPIRAGGDPGVAASNTGGGLLYSR
                     ESAPPPTAPFNLADINDVSPSGGFVAELLRPELDPVYIPPQQPQPPGGGLMGKFVLKAS
                     LTTPGSEYSSPSVISVSKGSPDGSHPVVVAPYSGGPPRMCPKIKQEAVPSCTVSRSLEA
                     HLSAGPQLSNGHRPNTHDFPLGRQLPTRTTPTLSPEELLNSRDCHPGLPLPPGFHPHPG
                     PNYPPFLPDQMQSQVPSLHYQELMPPGSCLPEEPKPKRGRRSWPRKRTATHTCDYAGCG
                     KTYTKSSHLKAHLRTHTGEKPYHCDWDGCGWKFARSDELTRHYRKHTGHRPFQCQKCDR
                     AFSRSDHLALHMKRHF"
     CDS             4357..4422
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             4447..5403
                     /codon_start=1
                     /label=mSox2
                     /note="Mus musculus transcription factor SOX-2 gene.
                     Belongs to the SRY-related HMG-box (SOX) family of 
                     transcription factors, which is involved in the regulation 
                     of embryonic development and in the determination of cell 
                     fate"
                     /translation="MYNMMETELKPPGPQQASGGGGGGGNATAAATGGNQKNSPDRVKR
                     PMNAFMVWSRGQRRKMAQENPKMHNSEISKRLGAEWKLLSETEKRPFIDEAKRLRALHM
                     KEHPDYKYRPRRKTKTLMKKDKYTLPGGLLAPGGNSMASGVGVGAGLGAGVNQRMDSYA
                     HMNGWSNGSYSMMQEQLGYPQHPGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGS
                     PTYSMSYSQQGTPGMALGSMGSVVKSEASSSPPVVTSSSHSRAPCQAGDLRDMISMYLP
                     GAEVPEPAAPSRLHMAQHYQSGPVPGTAINGTLPLSHM"
     primer_bind     complement(5516..5535)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5581..5636
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5635..5654)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also
                     called rb-glob-pA-term-R"
     primer_bind     complement(5997..6013)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6021..6037)
                     /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(6045..6075)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6090..6111)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6169..6365
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     polyA_signal    6371..6505
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6573..6590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6744..7332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7506..8363)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8364..8468)
                     /label=AmpR promoter
     promoter        8495
                     /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 ggcgaggggc
      541 ggggcgggcg aggcggagag gtgcggcggc 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 tcgcccttaa
     1741 aaagcaggct ccaccttccc catggctgga cacctggctt cagacttcgc cttctcaccc
     1801 ccaccaggtg ggggtgatgg gtcagcaggg ctggagccgg gctgggtgga ttctcgaacc
     1861 tggctaagct tccaagggcc tccaggtggg cctggaatcg gaccaggctc agaggtattg
     1921 gggatctccc catgtccgcc cgcatacgag ttctgcggag ggatggcata ctgtggacct
     1981 caggttggac tgggcctagt cccccaagtt ggcgtggaga ctttgcagcc tgagggccag
     2041 gcaggagcac gagtggaaag caactcagag ggaacctcct ctgagccctg tgccgaccgc
     2101 cccaatgccg tgaagttgga gaaggtggaa ccaactcccg aggagtccca ggacatgaaa
     2161 gccctgcaga aggagctaga acagtttgcc aagctgctga agcagaagag gatcaccttg
     2221 gggtacaccc aggccgacgt ggggctcacc ctgggcgttc tctttggaaa ggtgttcagc
     2281 cagaccacca tctgtcgctt cgaggccttg cagctcagcc ttaagaacat gtgtaagctg
     2341 cggcccctgc tggagaagtg ggtggaggaa gccgacaaca atgagaacct tcaggagata
     2401 tgcaaatcgg agaccctggt gcaggcccgg aagagaaagc gaactagcat tgagaaccgt
     2461 gtgaggtgga gtctggagac catgtttctg aagtgcccga agccctccct acagcagatc
     2521 actcacatcg ccaatcagct tgggctagag aaggatgtgg ttcgagtatg gttctgtaac
     2581 cggcgccaga agggcaaaag atcaagtatt gagtattccc aacgagaaga gtatgaggct
     2641 acagggacac ctttcccagg gggggctgta tcctttcctc tgcccccagg tccccacttt
     2701 ggcaccccag gctatggaag cccccacttc accacactct actcagtccc ttttcctgag
     2761 ggcgaggcct ttccctctgt tcccgtcact gctctgggct ctcccatgca ttcaaacggg
     2821 gatctcaaaa ttgtcgctcc tgtcaaacaa actcttaact ttgatttact caaactggct
     2881 ggggatgtag aaagcaatcc aggtccactc atggctgtca gcgacgctct gctcccgtcc
     2941 ttctccacgt tcgcgtccgg cccggcggga agggagaaga cactgcgtcc agcaggtgcc
     3001 ccgactaacc gttggcgtga ggaactctct cacatgaagc gacttccccc acttcccggc
     3061 cgcccctacg acctggcggc gacggtggcc acagacctgg agagtggcgg agctggtgca
     3121 gcttgcagca gtaacaaccc ggccctccta gcccggaggg agaccgagga gttcaacgac
     3181 ctcctggacc tagactttat cctttccaac tcgctaaccc accaggaatc ggtggccgcc
     3241 accgtgacca cctcggcgtc agcttcatcc tcgtcttccc cagcgagcag cggccctgcc
     3301 agcgcgccct ccacctgcag cttcagctat ccgatccggg ccgggggtga cccgggcgtg
     3361 gctgccagca acacaggtgg agggctcctc tacagccgag aatctgcgcc acctcccacg
     3421 gcccccttca acctggcgga catcaatgac gtgagcccct cgggcggctt cgtggctgag
     3481 ctcctgcggc cggagttgga cccagtatac attccgccac agcagcctca gccgccaggt
     3541 ggcgggctga tgggcaagtt tgtgctgaag gcgtctctga ccacccctgg cagcgagtac
     3601 agcagccctt cggtcatcag tgttagcaaa ggaagcccag acggcagcca ccccgtggta
     3661 gtggcgccct acagcggtgg cccgccgcgc atgtgcccca agattaagca agaggcggtc
     3721 ccgtcctgca cggtcagccg gtccctagag gcccatttga gcgctggacc ccagctcagc
     3781 aacggccacc ggcccaacac acacgacttc cccctggggc ggcagctccc caccaggact
     3841 acccctacac tgagtcccga ggaactgctg aacagcaggg actgtcaccc tggcctgcct
     3901 cttcccccag gattccatcc ccatccgggg cccaactacc ctcctttcct gccagaccag
     3961 atgcagtcac aagtcccctc tctccattat caagagctca tgccaccggg ttcctgcctg
     4021 ccagaggagc ccaagccaaa gaggggaaga aggtcgtggc cccggaaaag aacagccacc
     4081 cacacttgtg actatgcagg ctgtggcaaa acctatacca agagttctca tctcaaggca
     4141 cacctgcgaa ctcacacagg cgagaaacct taccactgtg actgggacgg ctgtgggtgg
     4201 aaattcgccc gctccgatga actgaccagg cactaccgca aacacacagg gcaccggccc
     4261 tttcagtgcc agaagtgcga cagggccttt tccaggtcgg accaccttgc cttacacatg
     4321 aagaggcact ttggggatct caaaattgtc gctcctgtca aacaaactct taactttgat
     4381 ttactcaaac tggctgggga tgtagaaagc aatccaggtc cactcatgaa caattcgccc
     4441 ttcaccatgt ataacatgat ggagacggag ctgaagccgc cgggcccgca gcaagcttcg
     4501 gggggcggcg gcggaggagg caacgccacg gcggcggcga ccggcggcaa ccagaagaac
     4561 agcccggacc gcgtcaagag gcccatgaac gccttcatgg tatggtcccg ggggcagcgg
     4621 cgtaagatgg cccaggagaa ccccaagatg cacaactcgg agatcagcaa gcgcctgggc
     4681 gcggagtgga aacttttgtc cgagaccgag aagcggccgt tcatcgacga ggccaagcgg
     4741 ctgcgcgctc tgcacatgaa ggagcacccg gattataaat accggccgcg gcggaaaacc
     4801 aagacgctca tgaagaagga taagtacacg cttcccggag gcttgctggc ccccggcggg
     4861 aacagcatgg cgagcggggt tggggtgggc gccggcctgg gtgcgggcgt gaaccagcgc
     4921 atggacagct acgcgcacat gaacggctgg agcaacggca gctacagcat gatgcaggag
     4981 cagctgggct acccgcagca cccgggcctc aacgctcacg gcgcggcaca gatgcaaccg
     5041 atgcaccgct acgacgtcag cgccctgcag tacaactcca tgaccagctc gcagacctac
     5101 atgaacggct cgcccaccta cagcatgtcc tactcgcagc agggcacccc cggtatggcg
     5161 ctgggctcca tgggctctgt ggtcaagtcc gaggccagct ccagcccccc cgtggttacc
     5221 tcttcctccc actccagggc gccctgccag gccggggacc tccgggacat gatcagcatg
     5281 tacctccccg gcgccgaggt gccggagccc gctgcgccca gtagactgca catggcccag
     5341 cactaccaga gcggcccggt gcccggcacg gccattaacg gcacactgcc cctgtcgcac
     5401 atgtgagggc tggaagggcg aattgcctgc aggaattcac tcctcaggtg caggctgcct
     5461 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     5521 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     5581 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     5641 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     5701 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     5761 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     5821 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     5881 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     5941 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
     6001 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6061 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6121 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     6181 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     6241 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6301 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     6361 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6421 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6481 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     6541 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6601 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6661 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6721 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6781 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6841 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6901 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6961 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7021 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7081 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7141 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7201 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7261 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7321 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7381 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7441 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7501 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7561 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7621 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7681 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7741 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7801 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7861 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7921 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7981 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8041 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8101 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8161 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8221 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8281 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8341 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8401 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8461 gttccgcgca catttccccg aaaagtgcca cctgg
//
