LOCUS       sequence-429278-        5053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express codon optimized Lin-28 and wild type Nanog. 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 5053)
  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 5053)
  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 339 CoMiP coLin28 2A Nanog IRES eGFP
FEATURES             Location/Qualifiers
     source          1..5053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        13..419
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     primer_bind     complement(545..562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(716..1304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(796..815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1794..1803
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2451..2516
                     /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"
     regulatory      4094..4103
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4100..4102,4103..4105,4106..4816)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4100..4102,4103..4105,4106..4816)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4145..4166)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4406..4425)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4753..4774
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4829..5053
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 aatctagctg cagtaacgcc attttgcaag gcatggaaaa ataccaaacc aagaatagag
       61 aagttcagat caagggcggg tacatgaaaa tagctaacgt tgggccaaac aggatatctg
      121 cggtgagcag tttcggcccc ggcccggggc caagaacaga tggtcaccgc agtttcggcc
      181 ccggcccgag gccaagaaca gatggtcccc agatatggcc caaccctcag cagtttctta
      241 agacccatca gatgtttcca ggctccccca aggacctgaa atgaccctgc gccttatttg
      301 aattaaccaa tcagcctgct tctcgcttct gttcgcgcgc ttctgcttcc cgagctctat
      361 aaaagagctc acaacccctc actcggcgcg ccagtcctcc gacagactga gtcggccggt
      421 ggatccaccg gtattgggat cttcacacag caggtaaggt tgcgggccgg gcctgggccg
      481 ggtccgggcc gggtattgcc cgcctaatga gcgggctttt ttttcttacc ccttcttccg
      541 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      601 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacgtgt
      661 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgccg gcgtttttcc
      721 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      781 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      841 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      901 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      961 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1021 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcggcc actggtaaca
     1081 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1141 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1201 gaaaaagagt tggtagctct tggtccggca aacaaaccac cgctggtagc ggtggttttt
     1261 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1321 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1381 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1441 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1501 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcctg caaaccacgt
     1561 tgtggtagaa ttggtaaaga gagtcgtgta aaatatcgag ttcgcacatc ttgttgtctg
     1621 attattgatt tttggcgaaa ccatttgatc atatgacaag atgtgtatct accttaactt
     1681 aatgattttg ataaaaatca ttaggtaccc cggcccgcac tgacccctgg tgttgctttt
     1741 tttttttagg ccgcaagctg aagcgtgtcc actagctgac tagctaaccg cgggccacca
     1801 tgggcagcgt gtccaaccag cagtttgccg gcggatgcgc caaggccgcc gaggaagctc
     1861 ctgaagaggc ccccgaagat gccgctagag ccgccgatga accccagctg ctgcatggcg
     1921 ccggaatctg caagtggttc aacgtgcgga tgggcttcgg cttcctgagc atgacagcca
     1981 gagccggcgt ggccctggac ccccctgtgg atgtgttcgt gcaccagagc aagctgcaca
     2041 tggaaggctt cagatccctg aaagagggcg aggccgtgga attcaccttc aagaagtccg
     2101 ccaagggcct ggaatccatc agagtgaccg gccctggcgg cgtgttctgc atcggcagcg
     2161 agagaaggcc caagggcaag agcatgcaga agcggcggag caagggcgac cggtgctaca
     2221 actgtggcgg cctggaccac cacgccaaag agtgcaagct gcctccccag cccaagaagt
     2281 gccacttctg ccagagcatc agccacatgg tggccagctg ccccctgaag gcccagcagg
     2341 gaccttctgc ccagggcaag cccacctact tcagagagga agaggaagag atccacagcc
     2401 ccaccctgct gcccgaggcc cagaacgggg atctcaaaat tgtcgctcct gtcaaacaaa
     2461 ctcttaactt tgatttactc aaactggctg gggatgtaga aagcaatcca ggtccactca
     2521 tgaacaattc gcccttcacc atgagtgtgg atccagcttg tccccaaagc ttgccttgct
     2581 ttgaagcatc cgactgtaaa gaatcttcac ctatgcctgt gatttgtggg cctgaagaaa
     2641 actatccatc cttgcaaatg tcttctgctg agatgcctca cacggagact gtctctcctc
     2701 ttccctcctc catggatctg cttattcagg acagccctga ttcttccacc agtcccaaag
     2761 gcaaacaacc cacttctgca gagaatagtg tcgcaaaaaa ggaagacaag gtcccagtca
     2821 agaaacagaa gaccagaact gtgttctctt ccacccagct gtgtgtactc aatgatagat
     2881 ttcagagaca gaaatacctc agcctccagc agatgcaaga actctccaac atcctgaacc
     2941 tcagctacaa acaggtgaag acctggttcc agaaccagag aatgaaatct aagaggtggc
     3001 agaaaaacaa ctggccgaag aatagcaatg gtgtgacgca gaaggcctca gcacctacct
     3061 accccagcct ctactcttcc taccaccagg gatgcctggt gaacccgact gggaaccttc
     3121 caatgtggag caaccagacc tggaacaatt caacctggag caaccagacc cagaacatcc
     3181 agtcctggag caaccactcc tggaacactc agacctggtg cacccaatcc tggaacaatc
     3241 aggcctggaa cagtcccttc tataactgtg gagaggaatc tctgcagtcc tgcatgcagt
     3301 tccagccaaa ttctcctgcc agtgacttgg aggctgcttt ggaagctgct ggggaaggcc
     3361 ttaatgtaat acagcagacc actaggtatt ttagtactcc acaaaccatg gatttattcc
     3421 taaactactc catgaacatg caacctgaag acgtgtgaaa ctagtggatc tccacgtggc
     3481 ggctagtact ccggtattgc ggtacccttg tacgcctgtt ttatactccc ttcccgtaac
     3541 ttagacgcac aaaaccaagt tcaatagaag ggggtacaaa ccagtaccac cacgaacaag
     3601 cacttctgtt tccccggtga tgtcgtatag actgcttgcg tggttgaaag cgacggatcc
     3661 gttatccgct tatgtacttc gagaagccca gtaccacctc ggaatcttcg atgcgttgcg
     3721 ctcagcactc aaccccagag tgtagcttag gctgatgagt ctggacatcc ctcaccggtg
     3781 acggtggtcc aggctgcgtt ggcggcctac ctatggctaa cgccatggga cgctagttgt
     3841 gaacaaggtg tgaagagcct attgagctac ataagaatcc tccggcccct gaatgcggct
     3901 aatcccaacc tcggagcagg tggtcacaaa ccagtgattg gcctgtcgta acgcgcaagt
     3961 ccgtggcgga accgactact ttgggtgtcc gtgtttcctt ttattttatt gtggctgctt
     4021 atggtgacaa tcacagattg ttatcataaa gcgaattgga taggatcaag cttatcgata
     4081 ccgtcgaggc cgcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     4141 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     4201 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     4261 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     4321 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4381 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4441 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4501 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     4561 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     4621 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4681 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4741 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4801 ggacgagctg tacaagaagc tttaatgact gtgccttcta gttgccagcc atctgttgtt
     4861 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4921 taaaatgagg aaattgcgtc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4981 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     5041 gtgggctcta tgg
//
