LOCUS       sequence-204154-        7946 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 7946)
  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 7946)
  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-AF9.643
FEATURES             Location/Qualifiers
     source          1..7946
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     25..44
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             113..712
                     /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(113..132)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     609..629
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    773..1340
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    790..1342
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(934..951)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1161..1180
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1354..1356,1357..1359,1360..2073)
                     /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(1399..1420)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1660..1679)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2007..2028
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2175..2689
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2851..2867
                     /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(2875..2881,2882..2899,2900..2905))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2920..2941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3058..3075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3229..3817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3309..3328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3988..4779,4780..4848))
                     /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     4611..4630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4849..4953)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5021..5039
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5077..5099)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5199..5218
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5412..5434
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5562..5581
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5781..6297
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6361..6702
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             6769..7185
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7115..7137
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7153..7169
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     ncRNA           7374..7388
                     /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(7607..7630)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc
       61 tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga
      121 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
      181 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
      241 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
      301 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
      361 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
      421 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
      481 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
      541 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
      601 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
      661 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
      721 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcca attccgcccc
      781 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
      841 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
      901 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
      961 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     1021 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     1081 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     1141 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     1201 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     1261 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     1321 tttgaaaaac acgatgataa tatggccaca accatggtga gcaagggcga ggagctgttc
     1381 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1441 gtgtctggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1501 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1561 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1621 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1681 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1741 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1801 aacgtctata tcatggccga caagcagaag aacggcatca aggcgaactt caagatccgc
     1861 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1921 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1981 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2041 atcactctcg gcatggacga gctgtacaag taatgaatta attaagaatt atcaagctta
     2101 tcgataccgt cgacctgcag ccaagcttat cgataaaata aaagatttta tttagtctcc
     2161 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     2221 cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg
     2281 aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     2341 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag
     2401 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     2461 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     2521 aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta
     2581 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     2641 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg ggtaacagtt
     2701 tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta atcctgactc
     2761 aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc attatggaca
     2821 gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc cacacaacat
     2881 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2941 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3001 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3061 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3121 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3181 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3241 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3301 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3361 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3421 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3481 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3541 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3601 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3661 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3721 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3781 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3841 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3901 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3961 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4021 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4081 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4141 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4201 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4261 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4321 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4381 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4441 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4501 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4561 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4621 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4681 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4741 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4801 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4861 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4921 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4981 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     5041 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     5101 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     5161 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     5221 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     5281 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5341 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     5401 gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag
     5461 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     5521 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     5581 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     5641 cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt tttgactcaa
     5701 caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt
     5761 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     5821 gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa
     5881 ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc
     5941 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt
     6001 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     6061 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     6121 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc
     6181 ccgggtaccc gtattcccaa taaagcctct tgctgtttgc atccgaatcg tggactcgct
     6241 gatccttggg agggtctcct cagattgatt gactgcccac ctcgggggtc tttcatttgg
     6301 aggttccacc gagatttgga gacccctgcc cagggaccac cgaccccccc gccgggaggt
     6361 aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc
     6421 taatgtttgc gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt
     6481 ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt
     6541 gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca
     6601 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
     6661 ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt
     6721 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt
     6781 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac
     6841 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc
     6901 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag
     6961 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc
     7021 tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct
     7081 ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg
     7141 cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggaat tagatctctc
     7201 gactagggat aacagggtaa ttgtttgaat gaggcttcag tactttacag aatcgttgcc
     7261 tgcacatctt ggaaacactt gctgggatta cttcgacttc ttaacccaac agaaggctcg
     7321 agaaggtata ttgctgttga cagtgagcga tggagataag atcaagaaga atagtgaagc
     7381 cacagatgta ttcttcttga tcttatctcc agtgcctact gcctcggact tcaaggggct
     7441 agaattcgag caattatctt gtttactaaa actgaatacc ttgctatctc tttgatacat
     7501 ttttacaaag ctgaattaaa atggtataaa ttaaatcact tttttcaatt gacgcgtgat
     7561 ctaattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc
     7621 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc
     7681 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc
     7741 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat
     7801 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg
     7861 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag
     7921 gctgggaagg ggtgggtccg ggggcg
//
