LOCUS       sequence-462996-        6737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of p53DD and HRAS(G12V) (DDIR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6737)
  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 6737)
  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: pMXs-p53DD-IRES-HRAS(G12V)
FEATURES             Location/Qualifiers
     source          1..6737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..133
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    222..246
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(738..755)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     965..984
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1134..1166,1167..1169,1170..1703)
                     /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(1718..1742)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1759..2347
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1812..1832)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2230..2241)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2527..2996
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3126..3143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3297..3885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3377..3396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4049..4840,4841..4909))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4672..4691
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4910..5014)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5044..5637
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5700..6057
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6116..6532
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6462..6484
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6500..6516
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(6542..6737,1..179)
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
ORIGIN
        1 tcggcgacac agtgtgggtc cgccgacacc agactaagaa cctagaacct cgctggaaag
       61 gaccttacac agtcctgctg accaccccca ccgccctcaa agtagacggc atcgcagctt
      121 ggatacacgc cgcccacgtg aaggctgccg accccggggg tggaccatcc tctagactgc
      181 cggatctagc tagttaatcg acggtatcga taagctatca aacaagtttg tacaaaaaag
      241 caggctccac catgggaacc aattcagtcg actggatccg gtaccatgac tgccatggag
      301 gagtcacagt cggatatcag cctcaagaga gcgctgccca cctgcacaag cgcctctccc
      361 ccgcaaaaga aaaaaccact tgatggagag tatttcaccc tcaagatccg cgggcgtaaa
      421 cgcttcgaga tgttccggga gctgaatgag gccttagagt taaaggatgc ccatgctaca
      481 gaggagtctg gagacagcag ggctcactcc agctacctga agaccaagaa gggccagtct
      541 acttcccgcc ataaaaaaac aatggtcaag aaagtggggc ctgactcaga ctaacgttac
      601 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
      661 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
      721 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
      781 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
      841 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
      901 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
      961 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     1021 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     1081 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaaatgaccg
     1141 aatacaagct ggtggtggtg ggcgccgtcg gtgtgggcaa gagtgcgctg accatccagc
     1201 tgatccagaa ccattttgtg gacgaatacg accccactat agaggattcc taccggaagc
     1261 aggtggtcat tgatggggag acgtgcctgt tggacatcct ggataccgcc ggccaggagg
     1321 agtacagcgc catgcgggac cagtacatgc gcaccgggga gggcttcctg tgtgtgtttg
     1381 ccatcaacaa caccaagtct tttgaggaca tccaccagta cagggagcag atcaaacggg
     1441 tgaaggactc ggatgacgtg cccatggtgc tggtggggaa caagtgtgac ctggctgcac
     1501 gcactgtgga atctcggcag gctcaggacc tcgcccgaag ctacggcatc ccctacatcg
     1561 agacctcggc caagacccgg cagggagtgg aggatgcctt ctacacgttg gtgcgtgaga
     1621 tccggcagca caagctgcgg aagctgaacc ctcctgatga gagtggcccc ggctgcatga
     1681 gctgcaagtg tgtgctctcc taactcgaga tatctagacc cagctttctt gtacaaagtg
     1741 gttcgatagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1801 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1861 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     1921 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     1981 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2041 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2101 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     2161 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     2221 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2281 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     2341 tccccgcatc gataccgtcg acgataaaat aaaagatttt atttagtctc cagaaaaagg
     2401 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     2461 aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg
     2521 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     2581 ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     2641 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt
     2701 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     2761 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     2821 ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg
     2881 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     2941 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcacatg
     3001 cagcatgtat caaaattaat ttggtttttt ttcttaagta tttacattaa atggccatag
     3061 ttgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3121 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3181 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3241 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3301 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3361 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3421 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3481 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3541 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3601 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3661 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3721 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3781 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3841 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3901 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3961 agattatcaa aaaggatctt cacctagatc cttttgcggc cggccgcaaa tcaatctaaa
     4021 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4081 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4141 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4201 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4261 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4321 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4381 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4441 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4501 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4561 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4621 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4681 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4741 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4801 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4861 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4921 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4981 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5041 cataatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
     5101 ggcatggaaa aatacataac tgagaataga aaagttcaga tcaaggtcag gaacagatgg
     5161 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     5221 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     5281 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
     5341 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     5401 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     5461 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgattg actgagtcgc
     5521 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     5581 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg
     5641 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta
     5701 agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg
     5761 cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact
     5821 gacgagttcg gaacacccgg ccgcaaccct gggagacgtc ccagggactt cgggggccgt
     5881 ttttgtggcc cgacctgagt ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc
     5941 ctaataggag ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc
     6001 gtctgaattt ttgctttcgg tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag
     6061 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca
     6121 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc
     6181 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg
     6241 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca
     6301 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat
     6361 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
     6421 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
     6481 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg cccccatatg
     6541 gccatatgag atcttatatg gggcaccccc gccccttgta aacttccctg accctgacat
     6601 gacaagagtt actaacagcc cctctctcca agctcactta caggctctct acttagtcca
     6661 gcacgaagtc tggagacctc tggcggcagc ctaccaagaa caactggacc gaccggtggt
     6721 acctcaccct taccgag
//
