LOCUS       sequence-438433-        9411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express Oct4, Klf4, Sox2, c-Myc and hairpin RNA p53. A single 
            plasmid reprogramming system using a mini-intronic plasmid (MIP)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9411)
  AUTHORS   Diecke S, Lu J, Lee J, Termglinchan V, Kooreman NG, Burridge PW, 
            Ebert AD, Churko JM, Sharma A, Kay MA, Wu JC
  TITLE     Novel codon-optimized mini-intronic plasmid for efficient, 
            inexpensive, and xeno-free induction of pluripotency.
  JOURNAL   Sci Rep. 2015 Jan 28;5:8081. doi: 10.1038/srep08081.
  PUBMED    25628230
REFERENCE   2  (bases 1 to 9411)
  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"; journalName: "Sci Rep."; 
            date: "2015-01-28"; volume: "5:8081. doi"; pages: " 
            10.1038/srep08081"
COMMENT     Sequence Label: MIP 247 CoMiP 4in1 with shRNA p53
FEATURES             Location/Qualifiers
     source          1..9411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        46..452
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     primer_bind     complement(578..595)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(749..1337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(829..848)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1827..1836
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2922..2978
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4401..4454
                     /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"
     CDS             5415..5474
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             5478..6794
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQSELQPPAPS
                     EDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTEL
                     LGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPNP
                     ARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDSL
                     LSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGSP
                     SAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNRK
                     CTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKAT
                     AYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
     misc_feature    join(6829..7403,7404..7406,7407..7415)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6996..7013)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7223..7242
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7416..8117
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(7549..7567)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7817..7836
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     8011..8034
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    8136..8724
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8189..8209)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8607..8618)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    8736..8960
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9118..9138)
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
ORIGIN
        1 ggctccccgg gcgcgactag tgaattcaga tctgagctag ctgcagtaac gccattttgc
       61 aaggcatgga aaaataccaa accaagaata gagaagttca gatcaagggc gggtacatga
      121 aaatagctaa cgttgggcca aacaggatat ctgcggtgag cagtttcggc cccggcccgg
      181 ggccaagaac agatggtcac cgcagtttcg gccccggccc gaggccaaga acagatggtc
      241 cccagatatg gcccaaccct cagcagtttc ttaagaccca tcagatgttt ccaggctccc
      301 ccaaggacct gaaatgaccc tgcgccttat ttgaattaac caatcagcct gcttctcgct
      361 tctgttcgcg cgcttctgct tcccgagctc tataaaagag ctcacaaccc ctcactcggc
      421 gcgccagtcc tccgacagac tgagtcggcc ggtggatcca ccggtattgg gatcttcaca
      481 cagcaggtaa ggttgcgggc cgggcctggg ccgggtccgg gccgggtatt gcccgcctaa
      541 tgagcgggct tttttttctt accccttctt ccgcttcctc gctcactgac tcgctgcgct
      601 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      661 cagaatcagg ggataacgca ggaaagaacg tgtgagcaaa aggccagcaa aaggccagga
      721 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      781 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      841 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      901 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
      961 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1021 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1081 acttatcgcc actggcagcg gccactggta acaggattag cagagcgagg tatgtaggcg
     1141 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1201 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttggtccg
     1261 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1321 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1381 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1441 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1501 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1561 catccatagt tgcctgactc ctgcaaacca cgttgtggta gaattggtaa agagagtcgt
     1621 gtaaaatatc gagttcgcac atcttgttgt ctgattattg atttttggcg aaaccatttg
     1681 atcatatgac aagatgtgta tctaccttaa cttaatgatt ttgataaaaa tcattaggta
     1741 ccccggcccg cactgacccc tggtgttgct tttttttttt aggccgcaag ctgaagcgtg
     1801 tccactagct gactagctaa accggtgcca ccatggccgg acacctggcc agcgacttcg
     1861 cctttagccc acctcctggc ggagggggag atggacctgg cggccctgaa cctggctggg
     1921 tggaccctcg gacctggctg tcctttcaag gccctccagg cggacctgga attggaccag
     1981 gcgtgggccc tggcagcgaa gtgtggggca tccctccctg ccctcccccc tacgagtttt
     2041 gcggcggcat ggcctactgt ggacctcagg tcggcgtggg actggtgcct cagggcggcc
     2101 tggaaacctc tcagcctgag ggcgaagccg gcgtgggcgt ggagagcaac tctgacggcg
     2161 ccagccctga gccttgtacc gtgaccccag gcgccgtgaa gctggaaaaa gagaagctgg
     2221 aacagaaccc cgaggaaagc caggacatca aggccctgca gaaagaactg gaacagttcg
     2281 ccaagctgct gaagcagaag cggatcaccc tgggctacac acaggctgac gtgggactga
     2341 cactgggcgt gctgttcggc aaggtgttca gccagaccac catctgcaga ttcgaagccc
     2401 tgcagctgtc cttcaagaac atgtgcaagc tgcggcccct gctgcagaaa tgggtggagg
     2461 aagccgacaa caacgagaac ctgcaggaaa tctgcaaggc cgagacactg gtgcaggccc
     2521 ggaagcggaa gcggaccagc atcgagaaca gagtgcgggg caacctggaa aacctgttcc
     2581 tgcagtgccc caagcccacc ctgcagcaga tcagccacat tgcacagcag ctgggactgg
     2641 aaaaggacgt cgtgagagtg tggttctgca accggcggca gaagggcaag agaagcagca
     2701 gcgactacgc ccagagagag gacttcgagg ccgctggctc cccttttagc ggcggacccg
     2761 tgtcctttcc tctggcccct ggccctcact ttggcacccc tggctacggc agcccacact
     2821 tcaccgccct gtacagcagc gtgccctttc ccgagggcga ggccttccct cctgtgtccg
     2881 tgaccaccct gggcagcccc atgcatagca actcttccgg agccaccaac ttctccctgc
     2941 tgaagcaggc cggcgacgtg gaggagaacc ccggcccctc catggctgtg tccgacgccc
     3001 tgctgcccag cttcagcacc tttgcctccg gccctgctgg cagagagaaa acactgcggc
     3061 aggctggcgc ccctaacaac agatggcggg aggaactgag ccacatgaag cggctgcctc
     3121 ccgtgctgcc cggcagacct tatgatctgg ccgctgccac cgtggccacc gatctggaat
     3181 ctggcggagc cggcgctgcc tgtggcggaa gcaacctggc ccccctgccc agacgggaga
     3241 cagaggaatt caacgacctg ctggacctgg acttcatcct gagcaacagc ctgacccacc
     3301 ctcctgagtc tgtggccgcc accgtgtcta gcagcgccag cgccagcagc agctctagcc
     3361 ctagcagcag cggacctgcc agcgccccca gcacctgcag cttcacctac cccatcagag
     3421 ccggcaatga ccctggcgtg gcccctggcg gaacaggagg aggactgctg tacggcagag
     3481 agtccgcccc tccccccacc gcccccttca acctggccga catcaacgat gtgtccccca
     3541 gcggcggctt tgtggccgag ctgctgcgac ctgagctgga ccccgtgtac atcccccctc
     3601 agcagccaca gcctcccggc ggaggactga tgggcaagtt cgtgctgaag gccagcctga
     3661 gcgcccctgg cagcgagtac ggcagcccca gcgtgatcag cgtgtccaag ggcagccctg
     3721 atggctctca ccctgtggtg gtggcccctt acaatggcgg ccctcccaga acctgcccca
     3781 agatcaagca ggaagccgtc agcagctgta cacacctggg agccggccct ccactgagca
     3841 atggccacag acctgccgcc cacgacttcc ctctgggaag gcagctccct agcagaacca
     3901 cccctaccct gggcctggaa gaggtgctgt ccagcagaga ctgccaccct gccctgcctc
     3961 tgccacctgg ctttcaccct caccctggcc ccaactaccc cagcttcctg cccgaccaga
     4021 tgcagcctca ggtgcccccc ctgcactacc aagaactgat gcccccaggc agctgcatgc
     4081 ccgaggaacc caagcccaag cggggcagaa gaagctggcc ccggaagaga accgccacac
     4141 acacctgcga ttacgccggc tgcggcaaga cctacaccaa gagcagccac ctgaaggccc
     4201 acctgagaac ccacaccggc gagaagccct accactgcga ctgggacggc tgtggctgga
     4261 agttcgccag aagcgacgag ctgacccggc actacagaaa gcacaccggc caccggccct
     4321 tccagtgcca gaagtgcgac cgggccttca gcagaagcga ccacctggct ctgcacatga
     4381 agagacactt cggctccgga gagggccggg gctctctgct gacctgtggc gacgtggagg
     4441 agaaccccgg cccctccatg tacaacatga tggaaaccga gctgaagccc cctggccctc
     4501 agcaaacatc tggcggcgga gggggcaatt ctacagccgc cgctgccggc ggaaaccaga
     4561 agaacagccc cgacagagtg aagcggccca tgaacgcctt catggtctgg tcccgcggcc
     4621 agcggcggaa gatggcccaa gaaaacccca agatgcacaa cagcgagatc agcaagagac
     4681 tgggcgccga gtggaagctg ctgtccgaga cagagaagag acccttcatc gacgaggcca
     4741 agcggctgag agccctgcac atgaaagagc accccgacta caagtaccgg cccagaagaa
     4801 agaccaagac cctgatgaag aaggacaagt acaccctgcc tggcggactg ctggcccctg
     4861 gcggcaattc tatggccagc ggagtgggag tgggagctgg actgggagcc ggcgtgaacc
     4921 agcggatgga cagctacgcc cacatgaacg gctggtccaa cggcagctac agcatgatgc
     4981 aggaccagct gggctaccct cagcaccctg gcctgaatgc ccatggcgcc gctcagatgc
     5041 agcccatgca cagatacgac gtgtccgccc tgcagtacaa cagcatgacc agcagccaga
     5101 cctacatgaa tggcagcccc acctacagca tgtcctacag ccagcagggc acacctggca
     5161 tggccctggg cagcatgggc agcgtggtca agagcgaggc cagcagcagc cctcctgtgg
     5221 tcaccagcag ctcccacagc agagcccctt gtcaggccgg cgacctgcgg gacatgatca
     5281 gcatgtacct gccaggcgcc gaagtgcctg aacctgccgc ccctagcaga ctgcacatga
     5341 gccagcacta ccagagcggc cctgtgcctg gcaccgccat caatggcacc ctgcccctga
     5401 gccacatgac gcgtcagtgc acaaactacg ccctgttgaa gctggctgga gacgtggagt
     5461 ccaaccctgg accctccatg cccctcaacg ttagcttcac caacaggaac tatgacctcg
     5521 actacgactc ggtgcagccg tatttctact gcgacgagga ggagaacttc taccagcagc
     5581 agcagagcga gctgcagccc ccggcgccca gcgaggatat ctggaagaaa ttcgagctgc
     5641 tgcccacccc gcccctgtcc cctagccgcc gctccgggct ctgctcgccc tcctacgttg
     5701 cggtcacacc cttctccctt cggggagaca acgacggcgg tggcgggagc ttctccacgg
     5761 ccgaccagct ggagatggtg accgagctgc tgggaggaga catggtgaac cagagtttca
     5821 tctgcgaccc ggacgacgag accttcatca aaaacatcat catccaggac tgtatgtgga
     5881 gcggcttctc ggccgccgcc aagctcgtct cagagaagct ggcctcctac caggctgcgc
     5941 gcaaagacag cggcagcccg aaccccgccc gcggccacag cgtctgctcc acctccagct
     6001 tgtacctgca ggatctgagc gccgccgcct cagagtgcat cgacccctcg gtggtcttcc
     6061 cctaccctct caacgacagc agctcgccca agtcctgcgc ctcgcaagac tccagcgcct
     6121 tctctccgtc ctcggattct ctgctctcct cgacggagtc ctccccgcag ggcagccccg
     6181 agcccctggt gctccatgag gagacaccgc ccaccaccag cagcgactct gaggaggaac
     6241 aagaagatga ggaagaaatc gatgttgttt ctgtggaaaa gaggcaggct cctggcaaaa
     6301 ggtcagagtc tggatcacct tctgctggag gccacagcaa acctcctcac agcccactgg
     6361 tcctcaagag gtgccacgtc tccacacatc agcacaacta cgcagcgcct ccctccactc
     6421 ggaaggacta tcctgctgcc aagagggtca agttggacag tgtcagagtc ctgagacaga
     6481 tcagcaacaa ccgaaaatgc accagcccca ggtcctcgga caccgaggag aatgtcaaga
     6541 ggcgaacaca caacgtcttg gagcgccaga ggaggaacga gctaaaacgg agcttttttg
     6601 ccctgcgtga ccagatcccg gagttggaaa acaatgaaaa ggcccccaag gtagttatcc
     6661 ttaaaaaagc cacagcatac atcctgtccg tccaagcaga ggagcaaaag ctcatttctg
     6721 aagaggactt gttgcggaaa cgacgagaac agttgaaaca caaacttgaa cagctacgga
     6781 actcttgtgc gtaagtcgac gaattcggta ccgcgggccc gggatccgcc cctctccctc
     6841 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     6901 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     6961 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     7021 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     7081 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     7141 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     7201 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     7261 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     7321 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     7381 cctttgaaaa acacgatgat aatatggcca caaccatggt gagcaagggc gaggaggtca
     7441 tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg
     7501 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     7561 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg
     7621 gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc
     7681 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga
     7741 cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca
     7801 acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg
     7861 agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga
     7921 aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc
     7981 aactgcccgg ctactactac gtggacacca agctggacat cacctcccac aacgaggact
     8041 acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc ctgtacggca
     8101 tggacgagct gtacaagtaa agcggccgcg tcgacaatca acctctggat tacaaaattt
     8161 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     8221 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     8281 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     8341 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     8401 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     8461 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     8521 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     8581 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     8641 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     8701 tctccctttg ggccgcctcc ccgcctggac tcgagctgtg ccttctagtt gccagccatc
     8761 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     8821 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     8881 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     8941 ggatgcggtg ggctctatgg agatctgata tctctagtcc cgggctgcag gaattccaaa
     9001 aaagactcca gtggtaatct actctcttga agtagattac cactggagtc ggcccaaaca
     9061 aggcttttct ccaagggata tttatagtct caaaacacac aattacttta cagttagggt
     9121 gagtttcctt ttgtgctgtt ttttaaaata ataatttagt atttgtatct cttatagaaa
     9181 tccaagccta tcatgtaaaa tgtagctagt attaaaaaga acagattatc tgtcttttat
     9241 cgcacattaa gcctctatag ttactaggaa atattatatg caaattaacc ggggcagggg
     9301 agtagccgag cttctcccac aagtctgtgc gagggggccg gcgcgggcct agagatggcg
     9361 gcgtcactag agtcgaccca tgggggcccg ccccaactgg ggtaaccttt g
//
