LOCUS       sequence-463023-        5889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of HRAS(G12V).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5889)
  AUTHORS   Wang NB, Lende-Dorn BA, Beitz AM, Han P, Adewumi HO, O'Shea TM, 
            Galloway KE
  TITLE     Proliferation history and transcription factor levels drive direct 
            conversion to motor neurons.
  JOURNAL   Cell Syst. 2025 Apr 16;16(4):101205. doi: 
            10.1016/j.cels.2025.101205. Epub 2025 Mar 13.
  PUBMED    40086434
REFERENCE   2  (bases 1 to 5889)
  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."; date: "2025-04-16"; volume: "16(4):101205. doi"; pages: " 
            10.1016/j.cels.2025.101205. Epub 2025 Mar 13"
COMMENT     Sequence Label: pMXs-HRAS(GV12)
FEATURES             Location/Qualifiers
     source          1..5889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(26..130)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             160..753
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    816..1173
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1232..1648
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1578..1600
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1616..1632
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1658..2032
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1970..1986
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    2075..2099
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(2139..2171,2172..2174,2175..2708)
                     /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(2723..2747)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2764..3352
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2817..2837)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3235..3246)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3532..4001
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4131..4148)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4302..4890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4382..4401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5054..5845,5846..5889,1..25))
                     /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     5677..5696
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
       61 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
      121 gggttccgcg cacatttccc cgaaaagtgc cacctgcata atgaaagacc ccacctgtag
      181 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag
      241 aatagaaaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag
      301 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
      361 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
      421 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
      481 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
      541 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
      601 cactcggcgc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
      661 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
      721 tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc
      781 cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt
      841 ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta ctagttagct
      901 aactagctct gtatctggcg gacccgtggt ggaactgacg agttcggaac acccggccgc
      961 aaccctggga gacgtcccag ggacttcggg ggccgttttt gtggcccgac ctgagtccaa
     1021 aaatcccgat cgttttggac tctttggtgc acccccctaa taggagggat atgtggttct
     1081 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
     1141 ggaccgaagc cgcgccgcgc gtcttgtctg ctgcagcatc gttctgtgtt gtctctgtct
     1201 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1261 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1321 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1381 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1441 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1501 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1561 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1621 tccagccctc actccttctc taggcgcccc catatggcca tatgagatct tatatggggc
     1681 acccccgccc cttgtaaact tccctgaccc tgacatgaca agagttacta acagcccctc
     1741 tctccaagct cacttacagg ctctctactt agtccagcac gaagtctgga gacctctggc
     1801 ggcagcctac caagaacaac tggaccgacc ggtggtacct cacccttacc gagtcggcga
     1861 cacagtgtgg gtccgccgac accagactaa gaacctagaa cctcgctgga aaggacctta
     1921 cacagtcctg ctgaccaccc ccaccgccct caaagtagac ggcatcgcag cttggataca
     1981 cgccgcccac gtgaaggctg ccgaccccgg gggtggacca tcctctagac tgccggatct
     2041 agctagttaa tcgacggtat cgataagcta tcaaacaagt ttgtacaaaa aagcaggctc
     2101 caccatggga accaattcag tcgactggat ccggtaccat gacggaatat aagctggtgg
     2161 tggtgggcgc cgtcggtgtg ggcaagagtg cgctgaccat ccagctgatc cagaaccatt
     2221 ttgtggacga atacgacccc actatagagg attcctaccg gaagcaggtg gtcattgatg
     2281 gggagacgtg cctgttggac atcctggata ccgccggcca ggaggagtac agcgccatgc
     2341 gggaccagta catgcgcacc ggggagggct tcctgtgtgt gtttgccatc aacaacacca
     2401 agtcttttga ggacatccac cagtacaggg agcagatcaa acgggtgaag gactcggatg
     2461 acgtgcccat ggtgctggtg gggaacaagt gtgacctggc tgcacgcact gtggaatctc
     2521 ggcaggctca ggacctcgcc cgaagctacg gcatccccta catcgagacc tcggccaaga
     2581 cccggcaggg agtggaggat gccttctaca cgttggtgcg tgagatccgg cagcacaagc
     2641 tgcggaagct gaaccctcct gatgagagtg gccccggctg catgagctgc aagtgtgtgc
     2701 tctcctgact cgagatatct agacccagct ttcttgtaca aagtggttcg atagcttatc
     2761 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2821 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2881 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2941 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3001 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3061 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3121 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     3181 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3241 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3301 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     3361 cgtcgacgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     3421 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     3481 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
     3541 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     3601 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     3661 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     3721 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     3781 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     3841 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     3901 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     3961 ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa
     4021 ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc
     4081 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4141 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4201 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4261 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4321 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4381 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4441 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4501 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4561 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4621 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4681 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4741 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4801 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4861 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4921 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4981 atcttcacct agatcctttt gcggccggcc gcaaatcaat ctaaagtata tatgagtaaa
     5041 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5101 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5161 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5221 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5281 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5341 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5401 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5461 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5521 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5581 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5641 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5701 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5761 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5821 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5881 gaataaggg
//
