LOCUS       sequence-204161-        7965 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  retrovirally express TADA1 shRNA with puro resistance and GFP marker
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7965)
  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 7965)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-shTADA1.1580
FEATURES             Location/Qualifiers
     source          1..7965
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     46..65
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             134..733
                     /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(134..153)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     630..650
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    794..1361
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    811..1363
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(955..972)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1182..1201
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1375..1377,1378..1380,1381..2094)
                     /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(1420..1441)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1681..1700)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2028..2049
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2196..2710
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2872..2888
                     /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(2896..2902,2903..2920,2921..2926))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2941..2962
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3079..3096)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3250..3838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3330..3349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4009..4800,4801..4869))
                     /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     4632..4651
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4870..4974)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5042..5060
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5098..5120)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5220..5239
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5433..5455
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5583..5602
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5802..6318
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6382..6723
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             6790..7206
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7136..7158
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7174..7190
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     ncRNA           7395..7409
                     /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(7628..7651)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
       61 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca
      121 gcccaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
      181 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
      241 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
      301 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
      361 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
      421 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
      481 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
      541 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
      601 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
      661 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
      721 gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc
      781 ccatgcatcc aattccgccc ccccccccta acgttactgg ccgaagccgc ttggaataag
      841 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
      901 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
      961 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     1021 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     1081 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     1141 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     1201 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     1261 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     1321 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg
     1381 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1441 gtaaacggcc acaagttcag cgtgtctggc gagggcgagg gcgatgccac ctacggcaag
     1501 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1561 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1621 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1681 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1741 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1801 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1861 aaggcgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1921 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1981 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2041 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatgaatt
     2101 aattaagaat tatcaagctt atcgataccg tcgacctgca gccaagctta tcgataaaat
     2161 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     2221 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     2281 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     2341 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     2401 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     2461 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     2521 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     2581 atagactgcg tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     2641 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     2701 ggtctttcat gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg
     2761 aatattaagt aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc
     2821 tgaaatagtt cattatggac agcgcagaaa gagctgggga gaattgtgaa attgttatcc
     2881 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2941 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3001 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3061 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3121 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3181 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3241 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3301 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3361 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3421 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3481 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3541 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3601 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3661 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3721 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3781 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3841 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3901 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3961 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4021 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4081 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4141 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4201 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4261 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4321 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4381 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4441 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4501 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4561 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4621 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4681 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4741 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4801 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4861 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4921 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4981 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5041 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     5101 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     5161 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     5221 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     5281 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     5341 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     5401 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     5461 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     5521 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     5581 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     5641 acgatgcgtc cggcgtagag gcgattagtc caatttgtta aagacaggat atcagtggtc
     5701 caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag
     5761 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     5821 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga
     5881 gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag
     5941 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     6001 gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     6061 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     6121 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag
     6181 tcctccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc ttgctgtttg
     6241 catccgaatc gtggactcgc tgatccttgg gagggtctcc tcagattgat tgactgccca
     6301 cctcgggggt ctttcatttg gaggttccac cgagatttgg agacccctgc ccagggacca
     6361 ccgacccccc cgccgggagg taagctggcc agcggtcgtt tcgtgtctgt ctctgtcttt
     6421 gtgcgtgttt gtgccggcat ctaatgtttg cgcctgcgtc tgtactagtt agctaactag
     6481 ctctgtatct ggcggacccg tggtggaact gacgagttct gaacacccgg ccgcaaccct
     6541 gggagacgtc ccagggactt tgggggccgt ttttgtggcc cgacctgagg aagggagtcg
     6601 atgtggaatc cgaccccgtc aggatatgtg gttctggtag gagacgagaa cctaaaacag
     6661 ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg
     6721 ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct
     6781 gaaaattagg gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat
     6841 gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc
     6901 tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga
     6961 gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac
     7021 caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag
     7081 ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt
     7141 gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta
     7201 ggcgccggaa ttagatctct cgactaggga taacagggta attgtttgaa tgaggcttca
     7261 gtactttaca gaatcgttgc ctgcacatct tggaaacact tgctgggatt acttcgactt
     7321 cttaacccaa cagaaggctc gagaaggtat attgctgttg acagtgagcg acactgtgtg
     7381 ttcagtgatg tatagtgaag ccacagatgt atacatcact gaacacacag tggtgcctac
     7441 tgcctcggac ttcaaggggc tagaattcga gcaattatct tgtttactaa aactgaatac
     7501 cttgctatct ctttgataca tttttacaaa gctgaattaa aatggtataa attaaatcac
     7561 ttttttcaat tgacgcgtga tctaattcta ccgggtaggg gaggcgcttt tcccaaggca
     7621 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
     7681 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
     7741 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
     7801 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
     7861 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
     7921 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc ggggg
//
