LOCUS       sequence-114454-        8963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express Oct4, Klf4, Sox2, and c-Myc. 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 8963)
  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 8963)
  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 227 CoMiP 4in1 wo shRNA p53
FEATURES             Location/Qualifiers
     source          1..8963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..436
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     primer_bind     complement(562..579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(733..1321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(813..832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1811..1820
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2906..2962
                     /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             4385..4438
                     /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             5399..5458
                     /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             5462..6781
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
                     SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
                     LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
                     PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
                     LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
                     PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
                     KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
                     TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
     misc_feature    join(6819..7393,7394..7396,7397..7405)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6986..7003)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7213..7232
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7406..8107
                     /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(7539..7557)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7807..7826
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     8001..8024
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    8126..8714
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8179..8199)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8597..8608)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    8726..8950
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 ctagtgaatt cagatctgag ctagctgcag taacgccatt ttgcaaggca tggaaaaata
       61 ccaaaccaag aatagagaag ttcagatcaa gggcgggtac atgaaaatag ctaacgttgg
      121 gccaaacagg atatctgcgg tgagcagttt cggccccggc ccggggccaa gaacagatgg
      181 tcaccgcagt ttcggccccg gcccgaggcc aagaacagat ggtccccaga tatggcccaa
      241 ccctcagcag tttcttaaga cccatcagat gtttccaggc tcccccaagg acctgaaatg
      301 accctgcgcc ttatttgaat taaccaatca gcctgcttct cgcttctgtt cgcgcgcttc
      361 tgcttcccga gctctataaa agagctcaca acccctcact cggcgcgcca gtcctccgac
      421 agactgagtc ggccggtgga tccaccggta ttgggatctt cacacagcag gtaaggttgc
      481 gggccgggcc tgggccgggt ccgggccggg tattgcccgc ctaatgagcg ggcttttttt
      541 tcttacccct tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
      601 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
      661 cgcaggaaag aacgtgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      721 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
      781 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
      841 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
      901 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
      961 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1021 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1081 agcggccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1141 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1201 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1261 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1321 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1381 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1441 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1501 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1561 actcctgcaa accacgttgt ggtagaattg gtaaagagag tcgtgtaaaa tatcgagttc
     1621 gcacatcttg ttgtctgatt attgattttt ggcgaaacca tttgatcata tgacaagatg
     1681 tgtatctacc ttaacttaat gattttgata aaaatcatta ggtaccccgg cccgcactga
     1741 cccctggtgt tgcttttttt ttttaggccg caagctgaag cgtgtccact agctgactag
     1801 ctaaaccggt gccaccatgg ccggacacct ggccagcgac ttcgccttta gcccacctcc
     1861 tggcggaggg ggagatggac ctggcggccc tgaacctggc tgggtggacc ctcggacctg
     1921 gctgtccttt caaggccctc caggcggacc tggaattgga ccaggcgtgg gccctggcag
     1981 cgaagtgtgg ggcatccctc cctgccctcc cccctacgag ttttgcggcg gcatggccta
     2041 ctgtggacct caggtcggcg tgggactggt gcctcagggc ggcctggaaa cctctcagcc
     2101 tgagggcgaa gccggcgtgg gcgtggagag caactctgac ggcgccagcc ctgagccttg
     2161 taccgtgacc ccaggcgccg tgaagctgga aaaagagaag ctggaacaga accccgagga
     2221 aagccaggac atcaaggccc tgcagaaaga actggaacag ttcgccaagc tgctgaagca
     2281 gaagcggatc accctgggct acacacaggc tgacgtggga ctgacactgg gcgtgctgtt
     2341 cggcaaggtg ttcagccaga ccaccatctg cagattcgaa gccctgcagc tgtccttcaa
     2401 gaacatgtgc aagctgcggc ccctgctgca gaaatgggtg gaggaagccg acaacaacga
     2461 gaacctgcag gaaatctgca aggccgagac actggtgcag gcccggaagc ggaagcggac
     2521 cagcatcgag aacagagtgc ggggcaacct ggaaaacctg ttcctgcagt gccccaagcc
     2581 caccctgcag cagatcagcc acattgcaca gcagctggga ctggaaaagg acgtcgtgag
     2641 agtgtggttc tgcaaccggc ggcagaaggg caagagaagc agcagcgact acgcccagag
     2701 agaggacttc gaggccgctg gctccccttt tagcggcgga cccgtgtcct ttcctctggc
     2761 ccctggccct cactttggca cccctggcta cggcagccca cacttcaccg ccctgtacag
     2821 cagcgtgccc tttcccgagg gcgaggcctt ccctcctgtg tccgtgacca ccctgggcag
     2881 ccccatgcat agcaactctt ccggagccac caacttctcc ctgctgaagc aggccggcga
     2941 cgtggaggag aaccccggcc cctccatggc tgtgtccgac gccctgctgc ccagcttcag
     3001 cacctttgcc tccggccctg ctggcagaga gaaaacactg cggcaggctg gcgcccctaa
     3061 caacagatgg cgggaggaac tgagccacat gaagcggctg cctcccgtgc tgcccggcag
     3121 accttatgat ctggccgctg ccaccgtggc caccgatctg gaatctggcg gagccggcgc
     3181 tgcctgtggc ggaagcaacc tggcccccct gcccagacgg gagacagagg aattcaacga
     3241 cctgctggac ctggacttca tcctgagcaa cagcctgacc caccctcctg agtctgtggc
     3301 cgccaccgtg tctagcagcg ccagcgccag cagcagctct agccctagca gcagcggacc
     3361 tgccagcgcc cccagcacct gcagcttcac ctaccccatc agagccggca atgaccctgg
     3421 cgtggcccct ggcggaacag gaggaggact gctgtacggc agagagtccg cccctccccc
     3481 caccgccccc ttcaacctgg ccgacatcaa cgatgtgtcc cccagcggcg gctttgtggc
     3541 cgagctgctg cgacctgagc tggaccccgt gtacatcccc cctcagcagc cacagcctcc
     3601 cggcggagga ctgatgggca agttcgtgct gaaggccagc ctgagcgccc ctggcagcga
     3661 gtacggcagc cccagcgtga tcagcgtgtc caagggcagc cctgatggct ctcaccctgt
     3721 ggtggtggcc ccttacaatg gcggccctcc cagaacctgc cccaagatca agcaggaagc
     3781 cgtcagcagc tgtacacacc tgggagccgg ccctccactg agcaatggcc acagacctgc
     3841 cgcccacgac ttccctctgg gaaggcagct ccctagcaga accaccccta ccctgggcct
     3901 ggaagaggtg ctgtccagca gagactgcca ccctgccctg cctctgccac ctggctttca
     3961 ccctcaccct ggccccaact accccagctt cctgcccgac cagatgcagc ctcaggtgcc
     4021 ccccctgcac taccaagaac tgatgccccc aggcagctgc atgcccgagg aacccaagcc
     4081 caagcggggc agaagaagct ggccccggaa gagaaccgcc acacacacct gcgattacgc
     4141 cggctgcggc aagacctaca ccaagagcag ccacctgaag gcccacctga gaacccacac
     4201 cggcgagaag ccctaccact gcgactggga cggctgtggc tggaagttcg ccagaagcga
     4261 cgagctgacc cggcactaca gaaagcacac cggccaccgg cccttccagt gccagaagtg
     4321 cgaccgggcc ttcagcagaa gcgaccacct ggctctgcac atgaagagac acttcggctc
     4381 cggagagggc cggggctctc tgctgacctg tggcgacgtg gaggagaacc ccggcccctc
     4441 catgtacaac atgatggaaa ccgagctgaa gccccctggc cctcagcaaa catctggcgg
     4501 cggagggggc aattctacag ccgccgctgc cggcggaaac cagaagaaca gccccgacag
     4561 agtgaagcgg cccatgaacg ccttcatggt ctggtcccgc ggccagcggc ggaagatggc
     4621 ccaagaaaac cccaagatgc acaacagcga gatcagcaag agactgggcg ccgagtggaa
     4681 gctgctgtcc gagacagaga agagaccctt catcgacgag gccaagcggc tgagagccct
     4741 gcacatgaaa gagcaccccg actacaagta ccggcccaga agaaagacca agaccctgat
     4801 gaagaaggac aagtacaccc tgcctggcgg actgctggcc cctggcggca attctatggc
     4861 cagcggagtg ggagtgggag ctggactggg agccggcgtg aaccagcgga tggacagcta
     4921 cgcccacatg aacggctggt ccaacggcag ctacagcatg atgcaggacc agctgggcta
     4981 ccctcagcac cctggcctga atgcccatgg cgccgctcag atgcagccca tgcacagata
     5041 cgacgtgtcc gccctgcagt acaacagcat gaccagcagc cagacctaca tgaatggcag
     5101 ccccacctac agcatgtcct acagccagca gggcacacct ggcatggccc tgggcagcat
     5161 gggcagcgtg gtcaagagcg aggccagcag cagccctcct gtggtcacca gcagctccca
     5221 cagcagagcc ccttgtcagg ccggcgacct gcgggacatg atcagcatgt acctgccagg
     5281 cgccgaagtg cctgaacctg ccgcccctag cagactgcac atgagccagc actaccagag
     5341 cggccctgtg cctggcaccg ccatcaatgg caccctgccc ctgagccaca tgacgcgtca
     5401 gtgcacaaac tacgccctgt tgaagctggc tggagacgtg gagtccaacc ctggaccctc
     5461 catgcccctc aacgttagct tcaccaacag gaactatgac ctcgactacg actcggtgca
     5521 gccgtatttc tactgcgacg aggaggagaa cttctaccag cagcagcagc agagcgagct
     5581 gcagcccccg gcgcccagcg aggatatctg gaagaaattc gagctgctgc ccaccccgcc
     5641 cctgtcccct agccgccgct ccgggctctg ctcgccctcc tacgttgcgg tcacaccctt
     5701 ctcccttcgg ggagacaacg acggcggtgg cgggagcttc tccacggccg accagctgga
     5761 gatggtgacc gagctgctgg gaggagacat ggtgaaccag agtttcatct gcgacccgga
     5821 cgacgagacc ttcatcaaaa acatcatcat ccaggactgt atgtggagcg gcttctcggc
     5881 cgccgccaag ctcgtctcag agaagctggc ctcctaccag gctgcgcgca aagacagcgg
     5941 cagcccgaac cccgcccgcg gccacagcgt ctgctccacc tccagcttgt acctgcagga
     6001 tctgagcgcc gccgcctcag agtgcatcga cccctcggtg gtcttcccct accctctcaa
     6061 cgacagcagc tcgcccaagt cctgcgcctc gcaagactcc agcgccttct ctccgtcctc
     6121 ggattctctg ctctcctcga cggagtcctc cccgcagggc agccccgagc ccctggtgct
     6181 ccatgaggag acaccgccca ccaccagcag cgactctgag gaggaacaag aagatgagga
     6241 agaaatcgat gttgtttctg tggaaaagag gcaggctcct ggcaaaaggt cagagtctgg
     6301 atcaccttct gctggaggcc acagcaaacc tcctcacagc ccactggtcc tcaagaggtg
     6361 ccacgtctcc acacatcagc acaactacgc agcgcctccc tccactcgga aggactatcc
     6421 tgctgccaag agggtcaagt tggacagtgt cagagtcctg agacagatca gcaacaaccg
     6481 aaaatgcacc agccccaggt cctcggacac cgaggagaat gtcaagaggc gaacacacaa
     6541 cgtcttggag cgccagagga ggaacgagct aaaacggagc ttttttgccc tgcgtgacca
     6601 gatcccggag ttggaaaaca atgaaaaggc ccccaaggta gttatcctta aaaaagccac
     6661 agcatacatc ctgtccgtcc aagcagagga gcaaaagctc atttctgaag aggacttgtt
     6721 gcggaaacga cgagaacagt tgaaacacaa acttgaacag ctacggaact cttgtgcgtg
     6781 atgagtcgac gaattcggta ccgcgggccc gggatccgcc cctctccctc ccccccccct
     6841 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     6901 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     6961 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     7021 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     7081 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     7141 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     7201 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     7261 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     7321 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     7381 acacgatgat aatatggcca caaccatggt gagcaagggc gaggaggtca tcaaagagtt
     7441 catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg
     7501 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     7561 cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc
     7621 gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt
     7681 caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc
     7741 ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc
     7801 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta
     7861 cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg
     7921 ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg
     7981 ctactactac gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt
     8041 ggaacagtac gagcgctccg agggccgcca ccacctgttc ctgtacggca tggacgagct
     8101 gtacaagtaa agcggccgcg tcgacaatca acctctggat tacaaaattt gtgaaagatt
     8161 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     8221 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     8281 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     8341 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     8401 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     8461 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     8521 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     8581 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     8641 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     8701 ggccgcctcc ccgcctggac tcgagctgtg ccttctagtt gccagccatc tgttgtttgc
     8761 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     8821 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     8881 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     8941 ggctctatgg agatctgata tct
//
