LOCUS       sequence-471184-        8676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NUP98::KDM5A in bicistronic retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8676)
  AUTHORS   Michmerhuizen NL, Heikamp EB, Iacobucci I, Umeda M, Arthur B, Mishra
            V, Park CS, Di Giacomo D, Hiltenbrand R, Gao Q, Radko-Juettner S, 
            Lott J, Martucci C, Subramanyam V, Hatton C, Wenge DV, Baviskar P, 
            Portola P, Claquin A, Chandra B, Baggett DW, Khalighifar A, Huang H,
            Zhou P, Long L, Shi H, Sun Y, Papachristou EK, Sekhar Reddy 
            Chilamakuri C, de Luna Vitorino FN, Gongora JM, Wu H, Pounds SB, 
            Janke LJ, Kentsis A, D'Santos CS, Garcia BA, Kriwacki RW, Chi H, 
            Klco JM, Armstrong SA, Mullighan CG
  TITLE     KAT6A and KAT7 Histone Acetyltransferase Complexes Are Molecular 
            Dependencies and Therapeutic Targets in NUP98-Rearranged Acute 
            Myeloid Leukemia.
  JOURNAL   Cancer Discov. 2025 Jun 19. doi: 10.1158/2159-8290.CD-24-1772.
  PUBMED    40536430
REFERENCE   2  (bases 1 to 8676)
  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"; journalName: "Cancer 
            Discov. 2025 Jun 19. doi: 10.1158/2159-8290.CD-24-1772."
COMMENT     Sequence Label: CL20_MSCV-NUP98::KDM5A-ires-GFP
FEATURES             Location/Qualifiers
     source          1..8676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     LTR             1129..1643
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(1745..2458,2459..2461,2462..2464))
                     /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(1790..1811)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2139..2158
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2398..2419
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2476..3028
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2478..3045
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2638..2657)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2867..2884
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(5309..5725)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(5325..5341)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(5357..5379)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5792..6133
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             6197..6713
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(6913..6932)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7060..7082)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7276..7295)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7395..7417
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7455..7473)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7541..7645
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7646..7714,7715..8506)
                     /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     complement(7864..7883)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa ctatttcagg
     1021 agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac ttaatattcg
     1081 actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat gaaagacccc
     1141 cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga gaccacaagt
     1201 cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg cagtctatcg
     1261 gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg ggagcagaag
     1321 cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca cagggtcatt
     1381 tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc tgagggcggg
     1441 accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg cttaccacag
     1501 atatcctgtt tggcccatat tctgctgtct ctctgttcct aaccttgatc tgaacttctc
     1561 tattctcagt tatgtatttt ccatgccttg caaaatggcg ttacttaagc tagcttgcca
     1621 aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa aatcttttat
     1681 tttatcgata agcttggctg caggtcgacg gtatcgataa gcttgataat tcttaattaa
     1741 ttcattactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     1801 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca
     1861 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     1921 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcgccttga
     1981 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
     2041 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     2101 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
     2161 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     2221 gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca
     2281 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
     2341 tgccgtaggt ggcatcgccc tcgccctcgc cagacacgct gaacttgtgg ccgtttacgt
     2401 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     2461 ccatggttgt ggccatatta tcatcgtgtt tttcaaagga aaaccacgtc cccgtggttc
     2521 ggggggccta gacgtttttt taacctcgac taaacacatg taaagcatgt gcaccgaggc
     2581 cccagatcag atcccataca atggggtacc ttctgggcat ccttcagccc cttgttgaat
     2641 acgcttgagg agagccattt gactctttcc acaactatcc aactcacaac gtggcactgg
     2701 ggttgtgccg cctttgcagg tgtatcttat acacgtggct tttggccgca gaggcacctg
     2761 tcgccaggtg gggggttccg ctgcctgcaa agggtcgcta cagacgttgt ttgtcttcaa
     2821 gaagcttcca gaggaactgc ttccttcacg acattcaaca gaccttgcat tcctttggcg
     2881 agaggggaaa gacccctagg aatgctcgtc aagaagacag ggccaggttt ccgggccctc
     2941 acattgccaa aagacggcaa tatggtggaa aataacatat agacaaacgc acaccggcct
     3001 tattccaagc ggcttcggcc agtaacgtta gggggggggg gcggaattcg cccttatgtc
     3061 ccaaactaac caagcatctg ctaactggtc tctttaagat cctccattgg tagtttgtag
     3121 ctcattatga aggaaggagg tggtgctgga cctgggctaa ctggcccctg cttctttgca
     3181 cagtttatac agatgtaatc ttcattttca gccatttctg gagatacacc cacacaaact
     3241 tgatgaaacc actcatcaca gccaccatca cattgtaccc agtctacctt gtccttgcag
     3301 ggcctttggc agttctgtgc tgcgcacaca gcattctcat catcagactc ctctgctcct
     3361 gaccagtcat actttgaggg gatgtccagc acctttttct ctcttttctt ttcagtgctc
     3421 tctttcacaa gttcaacttt ggctgcagca gccttctcct tctttttctt tctctcttct
     3481 tcttttgcta gtttcttggc cagtttattc agctcctttg atttgtctgc acctaatttt
     3541 aatttcttct ttctaggttt gtccattttc tttaactcct tggacttctg ttttccttct
     3601 ccaaaaagtt gctctacctt ttctagcttc cgtttccgtt tcttctctga agagtccttt
     3661 cctttcactt ttagtggttt ctcttccatg ctgtcatcct cttccttctt cttagggtct
     3721 gacatcggat tccggaagag aggagagtct ccaaaaggtg agtatgttag actattgatg
     3781 tgctgctgga gaacagcctg ctgggcagca gaagcatttg gatctgtcaa agctactggg
     3841 gcctgggggg ctccaaagcc caaagtggct gtcgtagtat taaatccagg ggccccaaag
     3901 gctcctgtac caagaggccc tccaatctta ggttggttgt tcccaaacaa agatgcctgt
     3961 ccagcaccaa gagctgttcc aaatcctgca ccaagcccag ttccaagagt cccaggtgct
     4021 ggtttacttc caaaaatact attcccactg gtatttgtgc caaaaccaaa gagcccgcca
     4081 ctggttgtac caaatgaagg tgcactggtt gtgctgcttc caaatgtagt aagcttgtta
     4141 ttgccaaaca gggtcgaacc aacagcacca aatccagtat tggtctgccc aaagagacct
     4201 gttcctgttc caaatgctgt cccagtgctg gtgtttgtag ctgtcccaaa aagacctcca
     4261 ggctgtgagg cttgggttac tccaaataat cccatggtgt tggtgcttgg ctgtcctatg
     4321 gtgctggtat taccaaagga aaagccagtg ttctgggtgg ttgtagcctg gccaaatggt
     4381 ttgctgaaga ggctggtagt ctgctgattc tgttggccaa agagaccacc tggatttgtt
     4441 ccaaatccag ttgtactagt tccaaaggca gttttgttct gaccatatgc aaagcctgaa
     4501 ttagtggtgg aggagctgaa gagtcctgtt gcgctggaag tggctggaga agacccaaac
     4561 aagccagttg tggtacctgc tcccacctgg ttctgtgggc ccttcctgtt agcctgataa
     4621 tcctctaaac gaagttcctc tagtgacttg ctttcatatt ctttcatagc agtaatacac
     4681 tggtgcttgg tacttatgtt agtgctaact ccagctttga ccatagtatc tgtaccagtt
     4741 ggagggttaa atttaatagt agtcccagta ggagcagctg taaaactact tggcccaaag
     4801 agggagccag atgtgctgcc aaaaggatta gaggtggtat ttgtggttcc aaagagtcct
     4861 ccactgctag tactggttcc aaaattgcca aagccagttg gtttattttg tgcaaaggca
     4921 ttgttttggg atgagaagag actggtccct gtgcttgcag ttccaaacaa ggtatttgct
     4981 gttcctgttg acgtaccaaa cccaaagcca gtgcttgtgg aggtagctgg ctggctaaat
     5041 gaactggttc caaacaatcc tcctggttta gtctgtgaat ttccaaagag gcctccagta
     5101 ttgttgctag aaccaaatgc agatgttcca aatgcccctc cactagtagt gccaaagcca
     5161 gtattctgtc caaatgttga agttgtgcca aagccacctg tgccaccccc aaagggtgtt
     5221 ccaaatgatt tgttaaacat cttcaaaatg agtctttgtt tcagaaagcg cgaagggcga
     5281 attcgttaac ctcgagagat ctaattccgg cgcctagaga aggagtgagg gctggataaa
     5341 gggaggatcg aggcggggtc gaacgaggag gttcaagggg gagagacggg gcggatggag
     5401 gaagaggagg cggaggctta gggtgtacaa agggcttgac ccagggaggg gggtcaaaag
     5461 ccaaggcttc ccaggtcacg atgtagggga cctggtctgg gtgtccatgc gggccaggtg
     5521 aaaagacctt gatcttaacc tgggtgatga ggtctcggtt aaaggtgccg tctcgcggcc
     5581 atccgacgtt aaaggttggc cattctgcag agcagaaggt aacccaacgt ctcttcttga
     5641 catctaccga ctggttgtga gcgatccgct cgacatcttt ccagtgacct aaggtcaaac
     5701 ttaagggagt ggtaacagtc tggccctaat tttcagacaa atacagaaac acagtcagac
     5761 agagacaaca cagaacgatg ctgcagcgct gcagcagaca agacgcgcgg cttcggttcc
     5821 aaaccgaaag caaaaattca gacggaggcg ggaactgttt taggttctcg tctcctacca
     5881 gaaccacata tcctgacggg gtcggattcc acatcgactc ccttcctcag gtcgggccac
     5941 aaaaacggcc cccaaagtcc ctgggacgtc tcccagggtt gcggccgggt gttcagaact
     6001 cgtcagttcc accacgggtc cgccagatac agagctagtt agctaactag tacagacgca
     6061 ggcgcaaaca ttagatgccg gcacaaacac gcacaaagac agagacagac acgaaacgac
     6121 cgctggccag cttacctccc ggcggggggg tcggtggtcc ctgggcaggg gtctccaaat
     6181 ctcggtggaa cctccaaatg aaagaccccc gaggtgggca gtcaatcaat ctgaggagac
     6241 cctcccaagg atcagcgagt ccacgattcg gatgcaaaca gcaagaggct ttattgggaa
     6301 tacgggtacc cgggcgacgc agtctatcgg aggactggcg cgccgagtga ggggttgtgg
     6361 gctcttttat tgagctcggg gagcagaagc gcgcgaacag aagcgagaag cgaactgatt
     6421 ggttagttca aataaggcac agggtcattt caggtccttg gggcaccctg gaaacatctg
     6481 atggttctct agaaactgct gagggcggga ccgcatctgg ggaccatctg ttcttggccc
     6541 tgagccgggg caggaactgc ttaccacaga tatcctgttt ggcccatatt ctgctgtctc
     6601 tctgttccta accttgatct gaacttctct attctcagtt atgtattttc catgccttgc
     6661 aaaatggcgt tacttaagct agcttgccaa acctacaggt ggggtctttc attcccccct
     6721 ttttctggag actaaataaa atcttttatt ttatctatgg ctcgtactct ataggcttca
     6781 gctggtgata ttgttgagtc aaaactagag cctggaccac tgatatcctg tctttaacaa
     6841 attggactaa tcgcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
     6901 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
     6961 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
     7021 tgggcgccat ctccttgcat gcaccattcc ttgcgccgtc gttttacaac gtcgtgactg
     7081 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     7141 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     7201 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     7261 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     7321 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     7381 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     7441 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     7501 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     7561 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     7621 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     7681 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     7741 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     7801 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     7861 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     7921 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     7981 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     8041 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     8101 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     8161 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     8221 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     8281 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     8341 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     8401 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     8461 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     8521 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     8581 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     8641 agcgtcagac cccgtagaaa agatcaaagg atcttc
//
