LOCUS       sequence-200617-        8107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retrovirally express TAF12 shRNA with Neo resistance and mCherry 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8107)
  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 8107)
  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: LENC-shTAF12.364
FEATURES             Location/Qualifiers
     source          1..8107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     ncRNA           1592..1606
                     /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"
     promoter        1790..2289
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1825..1848)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2222..2241
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2303..3106
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2366..2385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2976..2995
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    3112..3678
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3129..3680
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3273..3290)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3500..3519
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3692..4402
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3837..3855)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4105..4124
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             4469..4983
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5152..5168)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5152..5168)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5165..5187)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5176..5192
                     /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(5200..5206,5207..5224,5225..5230))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5245..5266
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5383..5400)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5554..6142)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5634..5653)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6313..7104,7105..7173))
                     /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     6936..6955
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7174..7278)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7346..7364
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7402..7424)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7524..7543
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7737..7759
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7887..7906
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8106..8107,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctctcga ctagggataa cagggtaatt
     1441 gtttgaatga ggcttcagta ctttacagaa tcgttgcctg cacatcttgg aaacacttgc
     1501 tgggattact tcgacttctt aacccaacag aaggctcgag aaggtatatt gctgttgaca
     1561 gtgagcgaca ggtattgacc aagaagaaat agtgaagcca cagatgtatt tcttcttggt
     1621 caatacctgg tgcctactgc ctcggacttc aaggggctag aattcgagca attatcttgt
     1681 ttactaaaac tgaatacctt gctatctctt tgatacattt ttacaaagct gaattaaaat
     1741 ggtataaatt aaatcacttt tttcaattga cgcgtgatct aattctaccg ggtaggggag
     1801 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
     1861 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
     1921 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
     1981 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
     2041 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     2101 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     2161 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
     2221 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     2281 ggcctttcga cctgcagcca atatgggatc ggccattgaa caagatggat tgcacgcagg
     2341 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2401 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2461 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     2521 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2581 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     2641 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     2701 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     2761 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     2821 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     2881 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     2941 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3001 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3061 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcaa ttccgccccc
     3121 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     3181 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     3241 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     3301 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     3361 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     3421 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     3481 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     3541 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     3601 tgtttagtcg aggttaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     3661 tgaaaaacac gatgataata tggccacaac catggtgagc aagggcgagg aggataacat
     3721 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     3781 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     3841 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     3901 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     3961 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     4021 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     4081 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     4141 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     4201 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     4261 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     4321 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     4381 catggacgag ctgtacaagt aagtcgacct gcagccaagc ttatcgataa aataaaagat
     4441 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     4501 agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc
     4561 agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa
     4621 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc
     4681 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     4741 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     4801 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact
     4861 gcgtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     4921 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     4981 catgggtaac agtttcttga agttggagaa caacattctg agggtaggag tcgaatatta
     5041 agtaatcctg actcaattag ccactgtttt gaatccacat actccaatac tcctgaaata
     5101 gttcattatg gacagcgcag aagagctggg gagaattaat tcgtaatcat ggtcatagct
     5161 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5221 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5281 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5341 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5401 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5461 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5521 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5581 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5641 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5701 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5761 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5821 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5881 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5941 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6001 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6061 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6121 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6181 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6241 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6301 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6361 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6421 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6481 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6541 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6601 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6661 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6721 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6781 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6841 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6901 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6961 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7021 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7081 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7141 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7201 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7261 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     7321 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     7381 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     7441 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     7501 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     7561 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     7621 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     7681 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     7741 gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac
     7801 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     7861 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     7921 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt
     7981 gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg
     8041 aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag
     8101 gggggaa
//
