LOCUS       sequence-10192-e        3035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3035)
  TITLE     shMDC1 plasmids
REFERENCE   2  (bases 1 to 3035)
  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"
COMMENT     Sequence Label: RasV12 in pENTR1A (w99-1)
FEATURES             Location/Qualifiers
     source          1..3035
                     /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(532..564,565..567,568..1101)
                     /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(1264..1363)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1475..1494)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1486..2295
                     /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
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1553..1572)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2385..2973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2874..2893
                     /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 gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatcccag tgtggtggta cagcttgggg ggccagggcc ggggccgagc gatgacggaa
      541 tataagctgg tggtggtggg cgccgtcggt gtgggcaaga gtgcgctgac catccagctg
      601 atccagaacc attttgtgga cgaatacgac cccactatag aggattccta ccggaagcag
      661 gtggtcattg atggggagac gtgcctgttg gacatcctgg ataccgccgg ccaggaggag
      721 tacagcgcca tgcgggacca gtacatgcgc accggggagg gcttcctgtg tgtgtttgcc
      781 atcaacaaca ccaagtcttt tgaggacatc caccagtaca gggagcagat caaacgggtg
      841 aaggactcgg atgacgtgcc catggtgctg gtggggaaca agtgtgacct ggctgcacgc
      901 actgtggaat ctcggcaggc tcaggacctc gcccgaagct acggcatccc ctacatcgag
      961 acctcggcca agacccggca gggagtggag gatgccttct acacgttggt gcgtgagatc
     1021 cggcagcaca agctgcggaa gctgaaccct cctgatgaga gtggccccgg ctgcatgagc
     1081 tgcaagtgtg tgctctcctg acgcagcaca agctcaggac atggaggtgc cggatgcagg
     1141 aaggaggtgc agacggaagg aggaggaagg aaggacggaa gcaaggaagg aaggaagggc
     1201 tgctggagcc cagtcacccc ccaagctgta ggaattcgcc agcacagtgg tcgagatatc
     1261 tagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     1321 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgcagctc
     1381 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     1441 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     1501 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     1561 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     1621 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     1681 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     1741 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     1801 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgtcc
     1861 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     1921 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     1981 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     2041 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     2101 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     2161 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     2221 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     2281 gatgagtttt tctaatcaga attggttaat tggttgtaac attattcaga ttgggccccg
     2341 ttccactgag cgtcagaccc ggtagaaaag atcaaaggat cttcttgaga tccttttttt
     2401 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2461 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2521 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2581 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2641 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2701 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2761 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2821 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2881 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2941 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3001 ttcctggcct tttgctggcc ttttgctcac atgtt
//
