LOCUS       sequence-203715-        7972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retrovirally express mouse TAF4 HFD with 3*FLAG at N terminal, GFP 
            marker and puro resistance, .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7972)
  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 7972)
  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: MSCV-PIG-FLAG-TAF4(827-909)
FEATURES             Location/Qualifiers
     source          1..7972
                     /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"
     CDS             1422..1442
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1443..1508
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        1783..2282
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1818..1841)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2215..2234
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2303..2902
                     /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(2303..2322)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2799..2819
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2963..3530
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2980..3532
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3124..3141)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3351..3370
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3544..3546,3547..3549,3550..4263)
                     /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(3589..3610)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3850..3869)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4197..4218
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4365..4879
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5041..5057
                     /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(5065..5071,5072..5089,5090..5095))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5110..5131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5248..5265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5419..6007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5499..5518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6178..6969,6970..7038))
                     /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     6801..6820
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7039..7143)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7211..7229
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7267..7289)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7389..7408
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7602..7624
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7752..7771
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7971..7972,1..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 gatctgccat gccaaagaag aagcgtaagg
     1441 tagattacaa agaccatgac ggtgattata aagatcatga tatcgattac aaggatgacg
     1501 atgacaaggg aggcctcgag gacaccttcc tcctccccgc gcctttgcag aggaggatac
     1561 tggagatagg taaaaagcac ggcatcacgg aactgcatcc agatgtagtg agttacgtgt
     1621 ctcatgccac acagcaaagg ctgcagaatc ttgtagaaaa aatatctgaa acggctcagc
     1681 agaaaaactt ctcttataag gacgacgaca gatacgagca agcgagtgat gtccgggctc
     1741 agctcaaatt ctttgaacaa ctggatcagt gagaattcta ccgggtaggg gaggcgcttt
     1801 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa
     1861 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct
     1921 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc
     1981 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc
     2041 agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc
     2101 agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc
     2161 tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct
     2221 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt
     2281 cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     2341 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     2401 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     2461 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     2521 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     2581 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     2641 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     2701 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     2761 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     2821 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     2881 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     2941 cgcacgaccc catgcatcca attccgcccc ccccccctaa cgttactggc cgaagccgct
     3001 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     3061 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     3121 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     3181 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     3241 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     3301 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     3361 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     3421 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     3481 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     3541 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3601 gacggcgacg taaacggcca caagttcagc gtgtctggcg agggcgaggg cgatgccacc
     3661 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     3721 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     3781 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3841 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3901 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3961 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     4021 aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     4081 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     4141 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     4201 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     4261 taatgaatta attaagaatt atcaagctta tcgataccgt cgacctgcag ccaagcttat
     4321 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     4381 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact
     4441 gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac
     4501 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     4561 atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     4621 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     4681 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
     4741 agtcctccga tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
     4801 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
     4861 gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac attctgaggg
     4921 taggagtcga atattaagta atcctgactc aattagccac tgttttgaat ccacatactc
     4981 caatactcct gaaatagttc attatggaca gcgcagaaag agctggggag aattgtgaaa
     5041 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5101 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5161 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5221 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5281 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5341 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5401 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5461 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5521 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5581 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5641 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5701 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5761 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5821 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5881 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5941 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6001 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6061 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6121 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6181 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6241 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6301 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6361 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6421 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6481 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6541 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6601 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6661 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6721 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6781 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6841 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6901 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6961 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7021 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7081 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7141 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     7201 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     7261 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     7321 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     7381 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     7441 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     7501 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     7561 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     7621 aacgacggcg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     7681 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     7741 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     7801 atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata
     7861 tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac
     7921 gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg aa
//
