LOCUS       sequence-471090-        7348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of HRAS(G12V) and SNAP-p53DD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7348)
  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 7348)
  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-SNAP-p53DD
FEATURES             Location/Qualifiers
     source          1..7348
                     /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(319..351,352..354,355..888)
                     /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(1045..1062)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1272..1291
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1446..1991
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     protein_bind    complement(2329..2353)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2370..2958
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2423..2443)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2841..2852)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3138..3607
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3737..3754)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3908..4496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3988..4007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4660..5451,5452..5520))
                     /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     5283..5302
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5521..5625)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5655..6248
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6311..6668
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6727..7143
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7073..7095
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7111..7127
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(7153..7348,1..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 gtactactga attcaagctt
      301 ggatcccaat gagccaccat gaccgaatac aagctggtgg tggtgggcgc cgtcggtgtg
      361 ggcaagagtg cgctgaccat ccagctgatc cagaaccatt ttgtggacga atacgacccc
      421 actatagagg attcctaccg gaagcaggtg gtcattgatg gggagacgtg cctgttggac
      481 atcctggata ccgccggcca ggaggagtac agcgccatgc gggaccagta catgcgcacc
      541 ggggagggct tcctgtgtgt gtttgccatc aacaacacca agtcttttga ggacatccac
      601 cagtacaggg agcagatcaa acgggtgaag gactcggatg acgtgcccat ggtgctggtg
      661 gggaacaagt gtgacctggc tgcacgcact gtggaatctc ggcaggctca ggacctcgcc
      721 cgaagctacg gcatccccta catcgaaacc tcggccaaga cccggcaggg agtggaggat
      781 gccttctaca cgttggtgcg tgagatccgg cagcacaagc tgcggaagct gaaccctcct
      841 gatgagagtg gccccggctg catgagctgc aagtgtgtgc tctcctaaca acaccatgga
      901 gcgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
      961 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     1021 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     1081 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     1141 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     1201 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     1261 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     1321 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     1381 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     1441 caggaatgga caaagactgc gaaatgaagc gcaccaccct ggatagccct ctgggcaagc
     1501 tggaactgtc tgggtgcgaa cagggcctgc accgtatcat cttcctgggc aaaggaacat
     1561 ctgccgccga cgccgtggaa gtgcctgccc cagccgccgt gctgggcgga ccagagccac
     1621 tgatgcaggc caccgcctgg ctcaacgcct actttcacca gcctgaggcc atcgaggagt
     1681 tccctgtgcc agccctgcac cacccagtgt tccagcagga gagctttacc cgccaggtgc
     1741 tgtggaaact gctgaaagtg gtgaagttcg gagaggtcat cagctacagc cacctggccg
     1801 ccctggccgg caatcccgcc gccaccgccg ccgtgaaaac cgccctgagc ggaaatcccg
     1861 tgcccattct gatcccctgc caccgggtgg tgcagggcga cctggacgtg gggggctacg
     1921 agggcgggct cgccgtgaaa gagtggctgc tggcccacga gggccacaga ctgggcaagc
     1981 ctgggctggg tcctgcaggc ggatccatga ctgccatgga ggagtcacag tcggatatca
     2041 gcctcaagag agcgctgccc acctgcacaa gcgcctctcc cccgcaaaag aaaaaaccac
     2101 ttgatggaga gtatttcacc ctcaagatcc gcgggcgtaa acgcttcgag atgttccggg
     2161 agctgaatga ggccttagag ttaaaggatg cccatgctac agaggagtct ggagacagca
     2221 gggctcactc cagctacctg aagaccaaga agggccagtc tacttcccgc cataaaaaaa
     2281 caatggtcaa gaaagtgggg cctgactcag actgacgctg atatctagac ccagctttct
     2341 tgtacaaagt ggttcgatag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     2401 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2461 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2521 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2581 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2641 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2701 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2761 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     2821 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2881 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2941 tttgggccgc ctccccgcat cgataccgtc gacgataaaa taaaagattt tatttagtct
     3001 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     3061 gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggtc
     3121 aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     3181 ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     3241 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag
     3301 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     3361 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     3421 tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat
     3481 tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
     3541 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
     3601 tctttcacat gcagcatgta tcaaaattaa tttggttttt tttcttaagt atttacatta
     3661 aatggccata gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3721 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3781 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3841 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3901 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3961 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4021 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4081 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4141 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4201 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4261 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4321 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4381 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4441 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4501 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4561 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttgcgg ccggccgcaa
     4621 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4681 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4741 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4801 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4861 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4921 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4981 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5041 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5101 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5161 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5221 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5281 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5341 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5401 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5461 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5521 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5581 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5641 agtgccacct gcataatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     5701 ccattttgca aggcatggaa aaatacataa ctgagaatag aaaagttcag atcaaggtca
     5761 ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     5821 cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     5881 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc
     5941 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     6001 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     6061 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgatt
     6121 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     6181 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     6241 ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc
     6301 accgggaggt aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga
     6361 ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc
     6421 gtggtggaac tgacgagttc ggaacacccg gccgcaaccc tgggagacgt cccagggact
     6481 tcgggggccg tttttgtggc ccgacctgag tccaaaaatc ccgatcgttt tggactcttt
     6541 ggtgcacccc cctaatagga gggatatgtg gttctggtag gagacgagaa cctaaaacag
     6601 ttcccgcctc cgtctgaatt tttgctttcg gtttgggacc gaagccgcgc cgcgcgtctt
     6661 gtctgctgca gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa
     6721 aattagggcc agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc
     6781 gagcggatcg ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc
     6841 tctgcagaat ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac
     6901 ctcatcaccc aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag
     6961 gtcccctaca tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc
     7021 tttgtacacc ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa
     7081 cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc
     7141 gcccccatat ggccatatga gatcttatat ggggcacccc cgccccttgt aaacttccct
     7201 gaccctgaca tgacaagagt tactaacagc ccctctctcc aagctcactt acaggctctc
     7261 tacttagtcc agcacgaagt ctggagacct ctggcggcag cctaccaaga acaactggac
     7321 cgaccggtgg tacctcaccc ttaccgag
//
