LOCUS       sequence-463010-        6743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of HRAS(G12V) and p53DD (RIDD).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6743)
  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 6743)
  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-HRAS(G12V)-IRES-p53DD
FEATURES             Location/Qualifiers
     source          1..6743
                     /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"
     CDS             join(286..318,319..321,322..855)
                     /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(1005..1022)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1232..1251
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     protein_bind    complement(1724..1748)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1765..2353
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1818..1838)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2236..2247)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2533..3002
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3132..3149)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3303..3891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3383..3402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4055..4846,4847..4915))
                     /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     4678..4697
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4916..5020)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5050..5643
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5706..6063
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6122..6538
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6468..6490
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6506..6522
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(6548..6743,1..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 cgaatacaag
      301 ctggtggtgg tgggcgccgt cggtgtgggc aagagtgcgc tgaccatcca gctgatccag
      361 aaccattttg tggacgaata cgaccccact atagaggatt cctaccggaa gcaggtggtc
      421 attgatgggg agacgtgcct gttggacatc ctggataccg ccggccagga ggagtacagc
      481 gccatgcggg accagtacat gcgcaccggg gagggcttcc tgtgtgtgtt tgccatcaac
      541 aacaccaagt cttttgagga catccaccag tacagggagc agatcaaacg ggtgaaggac
      601 tcggatgacg tgcccatggt gctggtgggg aacaagtgtg acctggctgc acgcactgtg
      661 gaatctcggc aggctcagga cctcgcccga agctacggca tcccctacat cgagacctcg
      721 gccaagaccc ggcagggagt ggaggatgcc ttctacacgt tggtgcgtga gatccggcag
      781 cacaagctgc ggaagctgaa ccctcctgat gagagtggcc ccggctgcat gagctgcaag
      841 tgtgtgctct cctaactcga gcgttactgg ccgaagccgc ttggaataag gccggtgtgc
      901 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
      961 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     1021 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     1081 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     1141 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     1201 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     1261 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     1321 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     1381 cgtggttttc ctttgaaaaa atgactgcca tggaggagtc acagtcggat atcagcctca
     1441 agagagcgct gcccacctgc acaagcgcct ctcccccgca aaagaaaaaa ccacttgatg
     1501 gagagtattt caccctcaag atccgcgggc gtaaacgctt cgagatgttc cgggagctga
     1561 atgaggcctt agagttaaag gatgcccatg ctacagagga gtctggagac agcagggctc
     1621 actccagcta cctgaagacc aagaagggcc agtctacttc ccgccataaa aaaacaatgg
     1681 tcaagaaagt ggggcctgac tcagactaac tcgagatatc tagacccagc tttcttgtac
     1741 aaagtggttc gatagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     1801 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     1861 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     1921 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     1981 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2041 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2101 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2161 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     2221 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2281 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2341 gccgcctccc cgcatcgata ccgtcgacga taaaataaaa gattttattt agtctccaga
     2401 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     2461 tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa
     2521 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     2581 gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     2641 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc
     2701 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     2761 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     2821 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
     2881 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     2941 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     3001 cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg
     3061 ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3121 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3181 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3241 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3301 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3361 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3421 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3481 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3541 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3601 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3661 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3721 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3781 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3841 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3901 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3961 gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccggc cgcaaatcaa
     4021 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4081 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4141 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4201 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4261 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4321 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4381 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4441 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4501 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4561 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4621 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4681 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4741 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4801 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4861 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4921 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4981 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5041 cacctgcata atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     5101 ttgcaaggca tggaaaaata cataactgag aatagaaaag ttcagatcaa ggtcaggaac
     5161 agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
     5221 ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     5281 cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca
     5341 gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
     5401 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
     5461 ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc cgattgactg
     5521 agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
     5581 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
     5641 atttgggggc tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg
     5701 gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat
     5761 tttatgcgcc tgcgtcggta ctagttagct aactagctct gtatctggcg gacccgtggt
     5821 ggaactgacg agttcggaac acccggccgc aaccctggga gacgtcccag ggacttcggg
     5881 ggccgttttt gtggcccgac ctgagtccaa aaatcccgat cgttttggac tctttggtgc
     5941 acccccctaa taggagggat atgtggttct ggtaggagac gagaacctaa aacagttccc
     6001 gcctccgtct gaatttttgc tttcggtttg ggaccgaagc cgcgccgcgc gtcttgtctg
     6061 ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta
     6121 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg
     6181 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc
     6241 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat
     6301 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc
     6361 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt
     6421 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc
     6481 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgcccc
     6541 catatggcca tatgagatct tatatggggc acccccgccc cttgtaaact tccctgaccc
     6601 tgacatgaca agagttacta acagcccctc tctccaagct cacttacagg ctctctactt
     6661 agtccagcac gaagtctgga gacctctggc ggcagcctac caagaacaac tggaccgacc
     6721 ggtggtacct cacccttacc gag
//
