LOCUS       sequence-463826-        3700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for p53DD and HRAS(G12V) (DDIR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3700)
  AUTHORS   Wang NB, Adewumi HO, Lende-Dorn BA, Beitz AM, O'Shea TM, Galloway KE
  TITLE     Compact transcription factor cassettes generate functional, 
            engraftable motor neurons by direct conversion.
  JOURNAL   Cell Syst. 2025 Mar 10:101206. doi: 10.1016/j.cels.2025.101206.
  PUBMED    40086435
REFERENCE   2  (bases 1 to 3700)
  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: "Cell Syst. 
            2025 Mar 10:101206. doi: 10.1016/j.cels.2025.101206."
COMMENT     Sequence Label: pENTR-p53DD-IRES-HRAS(G12V)
FEATURES             Location/Qualifiers
     source          1..3700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(949..966)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1176..1195
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1345..1377,1378..1380,1381..1914)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase H-Ras"
                     /label=H-Ras (G12V)
                     /translation="MTEYKLVVVGAVGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK
                     DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI
                     RQHKLRKLNPPDESGPGCMSCKCVLS"
     protein_bind    complement(1929..2028)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2140..2159)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2151..2960
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2218..2237)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3050..3638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3539..3558
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatgggaac caattcagtc
      481 gactggatcc ggtaccatga ctgccatgga ggagtcacag tcggatatca gcctcaagag
      541 agcgctgccc acctgcacaa gcgcctctcc cccgcaaaag aaaaaaccac ttgatggaga
      601 gtatttcacc ctcaagatcc gcgggcgtaa acgcttcgag atgttccggg agctgaatga
      661 ggccttagag ttaaaggatg cccatgctac agaggagtct ggagacagca gggctcactc
      721 cagctacctg aagaccaaga agggccagtc tacttcccgc cataaaaaaa caatggtcaa
      781 gaaagtgggg cctgactcag actaacgtta ctggccgaag ccgcttggaa taaggccggt
      841 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
      901 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
      961 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     1021 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     1081 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     1141 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     1201 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     1261 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     1321 gggacgtggt tttcctttga aaaaatgacc gaatacaagc tggtggtggt gggcgccgtc
     1381 ggtgtgggca agagtgcgct gaccatccag ctgatccaga accattttgt ggacgaatac
     1441 gaccccacta tagaggattc ctaccggaag caggtggtca ttgatgggga gacgtgcctg
     1501 ttggacatcc tggataccgc cggccaggag gagtacagcg ccatgcggga ccagtacatg
     1561 cgcaccgggg agggcttcct gtgtgtgttt gccatcaaca acaccaagtc ttttgaggac
     1621 atccaccagt acagggagca gatcaaacgg gtgaaggact cggatgacgt gcccatggtg
     1681 ctggtgggga acaagtgtga cctggctgca cgcactgtgg aatctcggca ggctcaggac
     1741 ctcgcccgaa gctacggcat cccctacatc gagacctcgg ccaagacccg gcagggagtg
     1801 gaggatgcct tctacacgtt ggtgcgtgag atccggcagc acaagctgcg gaagctgaac
     1861 cctcctgatg agagtggccc cggctgcatg agctgcaagt gtgtgctctc ctaactcgag
     1921 atatctagac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1981 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctgc
     2041 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     2101 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     2161 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     2221 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     2281 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     2341 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     2401 attttatccg tactcctggt gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     2461 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     2521 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     2581 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     2641 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     2701 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     2761 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     2821 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     2881 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     2941 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacattat tcagattggg
     3001 ccccgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3061 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3121 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3181 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3241 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3301 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3361 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3421 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3481 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3541 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3601 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3661 tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
