LOCUS       sequence-463020-        7174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of myristoylated Akt1 (myr-Akt) and Bcl2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7174)
  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 7174)
  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-myrAkt-P2A-BCL2
FEATURES             Location/Qualifiers
     source          1..7174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(100..204)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             234..827
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    890..1247
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1306..1722
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1652..1674
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1690..1706
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1732..2106
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2044..2060
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    2157..2181
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2191..2211
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3676..3702
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3721..3777
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     protein_bind    complement(4550..4574)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     LTR             4891..5360
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5490..5507)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5661..6249)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5741..5760)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6413..7174,1..30,31..99))
                     /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     7036..7055
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
       61 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
      121 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
      181 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg cataatgaaa
      241 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
      301 aatacataac tgagaataga aaagttcaga tcaaggtcag gaacagatgg aacagctgaa
      361 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
      421 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
      481 tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac
      541 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
      601 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
      661 agcccacaac ccctcactcg gcgcgccagt cctccgattg actgagtcgc ccgggtaccc
      721 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
      781 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc
      841 gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta agctggccag
      901 caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc
      961 ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttcg
     1021 gaacacccgg ccgcaaccct gggagacgtc ccagggactt cgggggccgt ttttgtggcc
     1081 cgacctgagt ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc ctaataggag
     1141 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
     1201 ttgctttcgg tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg
     1261 tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc
     1321 actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag
     1381 tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt
     1441 aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc
     1501 aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg
     1561 gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg
     1621 cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct
     1681 cgatcctccc tttatccagc cctcactcct tctctaggcg cccccatatg gccatatgag
     1741 atcttatatg gggcaccccc gccccttgta aacttccctg accctgacat gacaagagtt
     1801 actaacagcc cctctctcca agctcactta caggctctct acttagtcca gcacgaagtc
     1861 tggagacctc tggcggcagc ctaccaagaa caactggacc gaccggtggt acctcaccct
     1921 taccgagtcg gcgacacagt gtgggtccgc cgacaccaga ctaagaacct agaacctcgc
     1981 tggaaaggac cttacacagt cctgctgacc acccccaccg ccctcaaagt agacggcatc
     2041 gcagcttgga tacacgccgc ccacgtgaag gctgccgacc ccgggggtgg accatcctct
     2101 agactgccgg atctagctag ttaattaagg atcccagtgt ggtggtacgg gaaatcacaa
     2161 gtttgtacaa aaaagcaggc tccaccatgg atggggagca gcaagagcaa gcccaagtct
     2221 agaatgaacg acgtagccat tgtgaaggag ggctggctgc acaaacgagg ggaatatatt
     2281 aaaacctggc ggccacgcta cttcctcctc aagaacgatg gcacctttat tggctacaag
     2341 gaacggcctc aggatgtgga tcagcgagag tccccactca acaacttctc agtggcacaa
     2401 tgccagctga tgaagacaga gcggccaagg cccaacacct ttatcatccg ctgcctgcag
     2461 tggaccacag tcattgagcg caccttccat gtggaaacgc ctgaggagcg ggaagaatgg
     2521 gccaccgcca ttcagactgt ggccgatgga ctcaagaggc aggaagaaga gacgatggac
     2581 ttccgatcag gctcacccag tgacaactca ggggctgaag agatggaggt gtccctggcc
     2641 aagcccaagc accgtgtgac catgaacgag tttgagtacc tgaaactact gggcaagggc
     2701 acctttggga aagtgattct ggtgaaagag aaggccacag gccgctacta tgccatgaag
     2761 atcctcaaga aggaggtcat cgtcgccaag gatgaggttg cccacacgct tactgagaac
     2821 cgtgtcctgc agaactctag gcatcccttc cttacggccc tcaagtactc attccagacc
     2881 cacgaccgcc tctgctttgt catggagtat gccaacgggg gcgagctctt cttccacctg
     2941 tctcgagagc gcgtgttctc cgaggaccgg gcccgcttct atggtgcgga gattgtgtct
     3001 gccctggact acttgcactc cgagaagaac gtggtgtacc gggacctgaa gctggagaac
     3061 ctcatgctgg acaaggacgg gcacatcaag ataacggact tcgggctgtg caaggagggg
     3121 atcaaggatg gtgccactat gaagacattc tgcggaacgc cggagtacct ggcccctgag
     3181 gtgctggagg acaacgacta cggccgtgca gtggactggt gggggctggg cgtggtcatg
     3241 tatgagatga tgtgtggccg cctgcccttc tacaaccagg accacgagaa gctgttcgag
     3301 ctgatcctca tggaggagat ccgcttcccg cgcacactcg gccctgaggc caagtccctg
     3361 ctctccgggc tgctcaagaa ggaccctaca cagaggctcg gtgggggctc tgaggatgcc
     3421 aaggagatca tgcagcaccg gttctttgcc aacatcgtgt ggcaggatgt gtatgagaag
     3481 aagctgagcc cacctttcaa gccccaggtc acctctgaga ctgacaccag gtatttcgat
     3541 gaggagttca cagctcagat gatcaccatc acgccgcctg atcaagatga cagcatggag
     3601 tgtgtggaca gtgagcggag gccgcacttc ccccagttct tctactcagc cagtggcaca
     3661 gcccccgggg gaagctatcc ttatgacgtg cctgactatg ccgtcgacac cggttccgga
     3721 gccacgaact tctctctgtt aaagcaagca ggagacgtgg aagaaaaccc cggtcctagg
     3781 ctgcagatgg cgcacgctgg gagaacaggg tacgataacc gggagatagt gatgaagtac
     3841 atccattata agctgtcgca gaggggctac gagtgggatg cgggagatgt gggcgccgcg
     3901 cccccggggg ccgcccccgc accgggcatc ttctcctccc agcccgggca cacgccccat
     3961 ccagccgcat cccgggaccc ggtcgccagg acctcgccgc tgcagacccc ggctgccccc
     4021 ggcgccgccg cggggcctgc gctcagcccg gtgccacctg tggtccacct gaccctccgc
     4081 caggccggcg acgacttctc ccgccgctac cgccgcgact tcgccgagat gtccagccag
     4141 ctgcacctga cgcccttcac cgcgcgggga cgctttgcca cggtggtgga ggagctcttc
     4201 agggacgggg tgaactgggg gaggattgtg gccttctttg agttcggtgg ggtcatgtgt
     4261 gtggagagcg tcaaccggga gatgtcgccc ctggtggaca acatcgccct gtggatgact
     4321 gagtacctga accggcacct gcacacctgg atccaggata acggaggctg ggatgccttt
     4381 gtggaactgt acggccccag catgcggcct ctgtttgatt tctcctggct gtctctgaag
     4441 actctgctca gtttggccct ggtgggagct tgcatcaccc tgggtgccta tctgggccac
     4501 aagtgactgg atccggtacc gaattcgcgg ccgcactcga gatatctaga cccagctttc
     4561 ttgtacaaag tggtgatatt ccagctgagc gccggtcgct accattacca gttggtctgg
     4621 tgtcaaaaat aataataacc gggcaggcca tgtctgcccg tatttcgcgt aaggaaatcc
     4681 attatgtact atttaaactc gagcggccgc cagcacagtg gtcgacgata aaataaaaga
     4741 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     4801 tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt
     4861 tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     4921 taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa
     4981 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     5041 agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     5101 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
     5161 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg
     5221 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag
     5281 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
     5341 ccgtcagcgg gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta
     5401 agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc
     5461 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5521 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5581 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5641 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5701 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5761 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5821 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5881 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5941 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6001 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6061 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6121 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6181 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6241 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6301 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg
     6361 cggccggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6421 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6481 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6541 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6601 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6661 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6721 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6781 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6841 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6901 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6961 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7021 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7081 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7141 tgtaacccac tcgtgcaccc aactgatctt cagc
//
