LOCUS       sequence-204157-        7959 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  retrovirally express MLL-AF9 shRNA with puro resistance and GFP 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7959)
  AUTHORS   Xu Y, Milazzo JP, Somerville TDD, Tarumoto Y, Huang YH, Ostrander 
            EL, Wilkinson JE, Challen GA, Vakoc CR
  TITLE     A TFIID-SAGA Perturbation that Targets MYB and Suppresses Acute 
            Myeloid Leukemia.
  JOURNAL   Cancer Cell. 2018 Jan 8;33(1):13-28.e8. doi: 
            10.1016/j.ccell.2017.12.002.
  PUBMED    29316427
REFERENCE   2  (bases 1 to 7959)
  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.ccell.2017.12"; journalName: "Cancer Cell"; date: 
            "2018-01-8- 8"; volume: "33"; issue: "1"; pages: "13-28.e8"
COMMENT     Sequence Label: LEP-shMLL-AF9.1039
FEATURES             Location/Qualifiers
     source          1..7959
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     40..59
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             128..727
                     /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(128..147)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     624..644
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    788..1355
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    805..1357
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(949..966)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1176..1195
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1369..1371,1372..1374,1375..2088)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1414..1435)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1675..1694)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2022..2043
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2190..2704
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2866..2882
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2890..2896,2897..2914,2915..2920))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2935..2956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3073..3090)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3244..3832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3324..3343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4003..4794,4795..4863))
                     /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     4626..4645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4864..4968)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5036..5054
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5092..5114)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5214..5233
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5427..5449
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5577..5596
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5796..6312
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6376..6717
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             6784..7200
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7130..7152
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7168..7184
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     ncRNA           7389..7403
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     primer_bind     complement(7622..7645)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc
       61 gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa
      121 gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
      181 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
      241 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
      301 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
      361 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
      421 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
      481 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
      541 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
      601 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
      661 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
      721 tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc
      781 atccaattcc gccccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
      841 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
      901 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
      961 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     1021 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     1081 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     1141 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     1201 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     1261 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     1321 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag
     1381 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1441 ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg caagctgacc
     1501 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1561 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1621 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1681 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1741 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1801 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcg
     1861 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1921 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1981 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2041 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatg aattaattaa
     2101 gaattatcaa gcttatcgat accgtcgacc tgcagccaag cttatcgata aaataaaaga
     2161 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     2221 tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt
     2281 cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta
     2341 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct
     2401 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     2461 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     2521 ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac
     2581 tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg
     2641 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt
     2701 tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga gtcgaatatt
     2761 aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata ctcctgaaat
     2821 agttcattat ggacagcgca gaaagagctg gggagaattg tgaaattgtt atccgctcac
     2881 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2941 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3001 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3061 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3661 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4981 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     5041 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     5101 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     5161 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     5221 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     5281 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     5341 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     5401 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg
     5461 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     5521 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     5581 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     5641 cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct
     5701 ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa
     5761 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     5821 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag
     5881 agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc
     5941 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     6001 ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     6061 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     6121 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
     6181 gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg
     6241 aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg
     6301 gggtctttca tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc
     6361 cccccgccgg gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt
     6421 gtttgtgccg gcatctaatg tttgcgcctg cgtctgtact agttagctaa ctagctctgt
     6481 atctggcgga cccgtggtgg aactgacgag ttctgaacac ccggccgcaa ccctgggaga
     6541 cgtcccaggg actttggggg ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg
     6601 aatccgaccc cgtcaggata tgtggttctg gtaggagacg agaacctaaa acagttcccg
     6661 cctccgtctg aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag
     6721 cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat
     6781 tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag
     6841 cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct
     6901 gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc
     6961 atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc
     7021 ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt
     7081 gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct
     7141 cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc
     7201 ggaattagat ctctcgacta gggataacag ggtaattgtt tgaatgaggc ttcagtactt
     7261 tacagaatcg ttgcctgcac atcttggaaa cacttgctgg gattacttcg acttcttaac
     7321 ccaacagaag gctcgagaag gtatattgct gttgacagtg agcgatccac ttacaaaaga
     7381 agataatagt gaagccacag atgtattatc ttcttttgta agtggagtgc ctactgcctc
     7441 ggacttcaag gggctagaat tcgagcaatt atcttgttta ctaaaactga ataccttgct
     7501 atctctttga tacattttta caaagctgaa ttaaaatggt ataaattaaa tcactttttt
     7561 caattgacgc gtgatctaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     7621 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     7681 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     7741 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
     7801 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
     7861 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
     7921 tttctgggct cagaggctgg gaaggggtgg gtccggggg
//
