LOCUS       sequence-476287-        7709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for overexpression of human RUNX1 in mammalian cells. Contains 
            mutations that prevent phosphorylation at some sites of the 
            translated protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7709)
  AUTHORS   Kwon N, Lu YC, Thompson EN, Mancuso RI, Wang L, Zhang PX, Krause DS
  TITLE     CDK9 phosphorylates RUNX1 to promote megakaryocytic fate in 
            megakaryocytic-erythroid progenitors.
  JOURNAL   Blood. 2024 Oct 24;144(17):1800-1812. doi: 10.1182/blood.2024023963.
  PUBMED    39102635
REFERENCE   2  (bases 1 to 7709)
  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: "10.1182/blood"; 
            journalName: "Blood"; date: "2024-10-24- 24"; volume: "144"; issue: 
            "17"; pages: "1800-1812"
COMMENT     Sequence Label: pMSCV-RUNX1-4A
FEATURES             Location/Qualifiers
     source          1..7709
                     /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"
     protein_bind    1418..1442
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1454..1480
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(2844..2868)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        2883..3382
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2918..2941)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3315..3334
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3403..4002
                     /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(3403..3422)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3899..3919
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4102..4616
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4778..4794
                     /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(4802..4808,4809..4826,4827..4832))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4847..4868
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4985..5002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5156..5744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5236..5255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5915..6706,6707..6775))
                     /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     6538..6557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6776..6880)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6948..6966
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7004..7026)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7126..7145
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7339..7361
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7489..7508
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7708..7709,1..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 gatcatcaca agtttgtaca aaaaagcagg
     1441 cttcaccatg gcatacccat acgacgtgcc tgactacgcc tcccgtatcc ccgtagatgc
     1501 cagcacgagc cgccgcttca cgccgccttc caccgcgctg agcccaggca agatgagcga
     1561 ggcgttgccg ctgggcgccc cggacgccgg cgctgccctg gccggcaagc tgaggagcgg
     1621 cgaccgcagc atggtggagg tgctggccga ccacccgggc gagctggtgc gcaccgacag
     1681 ccccaacttc ctctgctccg tgctgcctac gcactggcgc tgcaacaaga ccctgcccat
     1741 cgctttcaag gtggtggccc taggggatgt tccagatggc actctggtca ctgtgatggc
     1801 tggcaatgat gaaaactact cggctgagct gagaaatgct accgcagcca tgaagaacca
     1861 ggttgcaaga tttaatgacc tcaggtttgt cggtcgaagt ggaagaggga aaagcttcac
     1921 tctgaccatc actgtcttca caaacccacc gcaagtcgcc acctaccaca gagccatcaa
     1981 aatcacagtg gatgggcccc gagaacctcg aagacatcgg cagaaactag atgatcggac
     2041 caagcccggg agcttgtcct tttccgagcg gctcagtgaa ctggagcagc tgcggcgcac
     2101 agccatgagg gtcagcccac accacccagc ccccacgccc aaccctcgtg cctccctgaa
     2161 ccactccact gcctttaacc ctcagcctca gagtcagatg caggatacaa ggcagatcca
     2221 accagcccca ccgtggtcct acgatcagtc ctaccaatac ctgggatcca ttgccgctcc
     2281 ttctgtgcac ccagcagcgc ccattgcacc tggacgtgcc agcggcatga caaccctctc
     2341 tgcagaactt tccagtcgac tctcaacggc acccgacctg acagcgttca gcgacccgcg
     2401 ccagttcccc gcgctgccct ccatctccga cccccgcatg cactatccag gcgccttcac
     2461 ctactccccg acgccggtca cctcgggcat cggcatcggc atgtcggcca tgggctcggc
     2521 cacgcgctac cacacctacc tgccgccgcc ctaccccggc tcgtcgcaag cgcagggagg
     2581 cccgttccaa gccagctcgc cctcctacca cctgtactac ggcgcctcgg ccggctccta
     2641 ccagttctcc atggtgggcg gcgagcgctc gccgccgcgc atcctgccgc cctgcaccaa
     2701 cgcctccacc ggctccgcgc tgctcaaccc cagcctcccg aaccagagcg acgtggtgga
     2761 ggccgagggc agccacagca actcccccac caacatggcg ccctccgcgc gcctggagga
     2821 ggccgtgtgg cggccctact gagacccagc tttcttgtac aaagtggtga tcgcgttcta
     2881 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
     2941 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
     3001 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
     3061 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
     3121 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
     3181 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
     3241 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc
     3301 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
     3361 ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc
     3421 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     3481 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     3541 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     3601 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     3661 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     3721 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     3781 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     3841 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     3901 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     3961 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg
     4021 cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa gattttattt
     4081 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     4141 gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa
     4201 ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc
     4261 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt
     4321 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     4381 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     4441 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc
     4501 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     4561 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatgggt
     4621 aacagtttct tgaagttgga gaacaacatt ctgagggtag gagtcgaata ttaagtaatc
     4681 ctgactcaat tagccactgt tttgaatcca catactccaa tactcctgaa atagttcatt
     4741 atggacagcg cagaaagagc tggggagaat tgtgaaattg ttatccgctc acaattccac
     4801 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4861 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4921 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4981 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5041 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5101 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5161 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5221 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5281 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5341 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5401 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5461 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5521 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5581 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5641 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5701 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5761 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5821 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5881 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5941 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6001 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6061 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6121 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6181 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6241 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6301 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6361 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6421 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6481 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6541 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6601 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6661 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6721 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6781 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6841 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6901 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     6961 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     7021 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     7081 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     7141 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     7201 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     7261 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     7321 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc
     7381 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     7441 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     7501 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     7561 gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt
     7621 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat
     7681 tttatttagt ctccagaaaa aggggggaa
//
