LOCUS       sequence-463831-        3706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for HRAS(G12V) and p53DD (RIDD).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3706)
  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 3706)
  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-HRAS(G12V)-IRES-p53DD
FEATURES             Location/Qualifiers
     source          1..3706
                     /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"
     CDS             join(497..529,530..532,533..1066)
                     /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"
     primer_bind     complement(1216..1233)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1443..1462
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     protein_bind    complement(1935..2034)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2146..2165)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2157..2966
                     /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(2224..2243)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3056..3644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3545..3564
                     /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 ccgaatacaa gctggtggtg gtgggcgccg tcggtgtggg
      541 caagagtgcg ctgaccatcc agctgatcca gaaccatttt gtggacgaat acgaccccac
      601 tatagaggat tcctaccgga agcaggtggt cattgatggg gagacgtgcc tgttggacat
      661 cctggatacc gccggccagg aggagtacag cgccatgcgg gaccagtaca tgcgcaccgg
      721 ggagggcttc ctgtgtgtgt ttgccatcaa caacaccaag tcttttgagg acatccacca
      781 gtacagggag cagatcaaac gggtgaagga ctcggatgac gtgcccatgg tgctggtggg
      841 gaacaagtgt gacctggctg cacgcactgt ggaatctcgg caggctcagg acctcgcccg
      901 aagctacggc atcccctaca tcgagacctc ggccaagacc cggcagggag tggaggatgc
      961 cttctacacg ttggtgcgtg agatccggca gcacaagctg cggaagctga accctcctga
     1021 tgagagtggc cccggctgca tgagctgcaa gtgtgtgctc tcctaactcg agcgttactg
     1081 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     1141 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     1201 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     1261 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     1321 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     1381 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     1441 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     1501 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     1561 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa aatgactgcc
     1621 atggaggagt cacagtcgga tatcagcctc aagagagcgc tgcccacctg cacaagcgcc
     1681 tctcccccgc aaaagaaaaa accacttgat ggagagtatt tcaccctcaa gatccgcggg
     1741 cgtaaacgct tcgagatgtt ccgggagctg aatgaggcct tagagttaaa ggatgcccat
     1801 gctacagagg agtctggaga cagcagggct cactccagct acctgaagac caagaagggc
     1861 cagtctactt cccgccataa aaaaacaatg gtcaagaaag tggggcctga ctcagactaa
     1921 ctcgagatat ctagacccag ctttcttgta caaagttggc attataagaa agcattgctt
     1981 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2041 agctgcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     2101 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     2161 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     2221 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     2281 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     2341 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     2401 tcaagcattt tatccgtact cctggtgatg catggttact caccactgcg atccccggaa
     2461 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     2521 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     2581 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     2641 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     2701 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     2761 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     2821 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     2881 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     2941 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cattattcag
     3001 attgggcccc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3061 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3121 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3181 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3541 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3601 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3661 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
