LOCUS       sequence-391639-        6307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Ascl1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6307)
  AUTHORS   Wang H, Keepers B, Qian Y, Xie Y, Colon M, Liu J, Qian L
  TITLE     Cross-lineage potential of Ascl1 uncovered by comparing diverse 
            reprogramming regulatomes.
  JOURNAL   Cell Stem Cell. 2022 Oct 6;29(10):1491-1504.e9. doi: 
            10.1016/j.stem.2022.09.006.
  PUBMED    36206732
REFERENCE   2  (bases 1 to 6307)
  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"; doi: 
            "10.1016/j.stem.2022.09"; journalName: "Cell Stem Cell"; date: 
            "2022-10-6- 6"; volume: "29"; issue: "10"; pages: "1491-1504.e9"
COMMENT     Sequence Label: pMX-Ascl1-puro
FEATURES             Location/Qualifiers
     source          1..6307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             24..617
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    680..1037
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1096..1512
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1442..1464
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1480..1496
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1522..1896
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1834..1850
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1919..1928
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2644..2664)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2649..2978
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2829..2964
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2988..3587
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2988..3007)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3484..3504
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3814..4283
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4413..4430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4584..5172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4664..4683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5336..6127,6128..6196))
                     /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     5959..5978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6197..6301)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tccccgaaaa gtgccacctg cataatgaaa gaccccacct gtaggtttgg caagctagct
       61 taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga aaagttcaga
      121 tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
      181 agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg
      241 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
      301 cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
      361 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
      421 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt
      481 cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
      541 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
      601 agcgggggtc tttcatttgg gggctcgtcc gggatcggga gacccctgcc cagggaccac
      661 cgacccacca ccgggaggta agctggccag caacttatct gtgtctgtcc gattgtctag
      721 tgtctatgac tgattttatg cgcctgcgtc ggtactagtt agctaactag ctctgtatct
      781 ggcggacccg tggtggaact gacgagttcg gaacacccgg ccgcaaccct gggagacgtc
      841 ccagggactt cgggggccgt ttttgtggcc cgacctgagt ccaaaaatcc cgatcgtttt
      901 ggactctttg gtgcaccccc ctaataggag ggatatgtgg ttctggtagg agacgagaac
      961 ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttgggaccg aagccgcgcc
     1021 gcgcgtcttg tctgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat
     1081 ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg
     1141 aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt
     1201 accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt
     1261 aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac
     1321 ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg
     1381 gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc
     1441 ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct
     1501 tctctaggcg cccccatatg gccatatgag atcttatatg gggcaccccc gccccttgta
     1561 aacttccctg accctgacat gacaagagtt actaacagcc cctctctcca agctcactta
     1621 caggctctct acttagtcca gcacgaagtc tggagacctc tggcggcagc ctaccaagaa
     1681 caactggacc gaccggtggt acctcaccct taccgagtcg gcgacacagt gtgggtccgc
     1741 cgacaccaga ctaagaacct agaacctcgc tggaaaggac cttacacagt cctgctgacc
     1801 acccccaccg ccctcaaagt agacggcatc gcagcttgga tacacgccgc ccacgtgaag
     1861 gctgccgacc ccgggggtgg accatcctct agactgccgg atctagctag ttaattaagc
     1921 caccatggag agctctggca agatggagag tggagccggc cagcagccgc agcccccgca
     1981 gcccttcctg cctcccgcag cctgcttctt tgcgaccgcg gcggcggcgg cagcggcggc
     2041 ggccgcggca gctcagagcg cgcagcagca acagccgcag gcgccgccgc agcaggcgcc
     2101 gcagctgagc ccggtggccg acagccagcc ctcagggggc ggtcacaagt cagcggccaa
     2161 gcaggtcaag cgccagcgct cgtcctctcc ggaactgatg cgctgcaaac gccggctcaa
     2221 cttcagcggc ttcggctaca gcctgccaca gcagcagccg gccgccgtgg cgcgccgcaa
     2281 cgagcgcgag cgcaaccggg tcaagttggt caacctgggc tttgccaccc tccgggagca
     2341 tgtccccaac ggcgcggcca acaagaagat gagcaaggtg gagacgctgc gctcggcggt
     2401 cgagtacatc cgcgcgctgc agcagctgct ggacgagcac gacgcggtga gcgctgcctt
     2461 tcaggcgggc gtcctgtcgc ccaccatctc ccccaactac tccaacgact tgaactctat
     2521 ggcgggttct ccggtctcgt cctactcctc cgacgaggga tcctacgacc ctcttagccc
     2581 agaggaacaa gagctgctgg actttaccaa ctggttctga ccagcacagt ggtcgacagc
     2641 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2701 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2761 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2821 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2881 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     2941 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttaccatg accgagtaca
     3001 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     3061 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     3121 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     3181 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     3241 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     3301 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     3361 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     3421 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     3481 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     3541 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
     3601 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcgtcgacg ataaaataaa
     3661 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     3721 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     3781 agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg
     3841 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc
     3901 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     3961 cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     4021 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     4081 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg
     4141 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
     4201 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     4261 tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg gttttttttc
     4321 ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg cgcggggaga
     4381 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4441 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4501 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4561 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4621 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4681 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4741 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4801 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4861 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4921 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4981 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5041 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5101 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5161 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5221 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5281 ttgcggccgg ccgcaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5341 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5401 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5461 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5521 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5581 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5641 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5701 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5761 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5821 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5881 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5941 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6001 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6061 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6121 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6181 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6241 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6301 gcacatt
//
