LOCUS       sequence-382234-        8896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of a mutated (mut, R42A), non-functional 
            version of the mitosis degradation (MD) domain of cyclin B1 fused to
            mouse Gata2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8896)
  AUTHORS   Silverio-Alves R, Kurochkin I, Rydstrom A, Vazquez Echegaray C, 
            Haider J, Nicholls M, Rode C, Thelaus L, Lindgren AY, Ferreira AG, 
            Brandao R, Larsson J, de Bruijn MFTR, Martin-Gonzalez J, Pereira CF
  TITLE     GATA2 mitotic bookmarking is required for definitive haematopoiesis.
  JOURNAL   Nat Commun. 2023 Aug 14;14(1):4645. doi: 10.1038/s41467-023-40391-x.
  PUBMED    37580379
REFERENCE   2  (bases 1 to 8896)
  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: "Nat 
            Commun."; date: "2023-08-14"; volume: "14(1):4645. doi"; pages: " 
            10.1038/s41467-023-40391-x"
COMMENT     Sequence Label: FUW-MDmut-Gata2
FEATURES             Location/Qualifiers
     source          1..8896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(17..36)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        208..587
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        589..787
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     742..762
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             805..985
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1032..1157
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1650..1883
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2068..2112
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2410..2527
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2592..3803
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3640..3659
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     misc_feature    5532..6120
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5585..5605)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6003..6014)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6123..6139)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6124..6140)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6645..6825
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6851..6868)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6857..6968
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(7114..7702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7194..7213)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7873..8664,8665..8733))
                     /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     8496..8515
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8734..8838)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
       61 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
      121 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
      181 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
      241 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      301 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      361 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      421 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      481 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      541 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      601 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      661 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      721 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      781 taagcagcgc gttttgcctg tactgggtct ctctggttag accagatctg agcctgggag
      841 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
      901 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
      961 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     1021 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     1081 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     1141 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1201 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1261 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     1321 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     1381 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     1441 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     1501 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     1561 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     1621 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     1681 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     1741 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     1801 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     1861 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     1921 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     1981 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     2041 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     2101 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     2161 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     2221 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     2281 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     2341 attcgattag tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt
     2401 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     2461 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     2521 aaattttcgg gtttattaca gggacagcag agatccagtt tggttaatta acccgtgtcg
     2581 gctccagatc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcctcac
     2641 ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt cctgatcctt ccgcccggac
     2701 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     2761 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     2821 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     2881 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     2941 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     3001 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     3061 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     3121 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     3181 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     3241 cttgaggcct tcgctaatgc gggaaagctc ttattcgggt gagatgggct ggggcaccat
     3301 ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg
     3361 ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc
     3421 ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccacctgc
     3481 cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg
     3541 ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt
     3601 ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta
     3661 tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat
     3721 catttgggtc aatatgtaat tttcagtgtt agactagtaa attgtccgct aaattctggc
     3781 cgtttttggc ttttttgtta gacgaagctt gggcccggga tccatggctg agaacaaggc
     3841 aaaagtgagc atggcaggag ccaagagggt gcccgtcacc gtgacagccg cttccaaacc
     3901 aggcctgagg cccgcaacag cactcggtga catcggcaat aaggtctctg aggaactgca
     3961 ggcccgggtg cctctcaagc gagaagctaa aactctgggc accggaaagg ggaccgtcaa
     4021 agctctcccc aagcctgtgg agaaagtccc tgtgtgcgaa ccagaggtgg cgcctgagca
     4081 gcctcgctgg atggcgcacc ccgccgtatt gaatgcgcag caccccgact cgcaccatcc
     4141 gggcctggcg cataactaca tggagccagc acagctgctg cctcccgacg aggtggatgt
     4201 cttcttcaac catctcgact cgcagggcaa cccttactac gccaacccgg cccacgcgcg
     4261 cgcgcgcgtt tcctacagcc cggcgcatgc ccgtctcacc ggaggccaga tgtgccgacc
     4321 acacttgttg cacagcccag gcttgccgtg gctggacggg ggcaaagcag ctctctctgc
     4381 cgccgctgcc catcaccaca gtccctggac cgtcagcccg ttctccaaga ccccgctgca
     4441 cccctcagct gctggagcac ccggagggcc tctgtctgtt tacccagggg ctgcgggtgg
     4501 gagcggggga ggcagtggga gctccgtggc ctccctcacc cccactgcag cccactcggg
     4561 ctcccatctc ttcggcttcc cacccacgcc acccaaagaa gtgtctccag accccagcac
     4621 aacaggagct gcttccccgg cctcttcttc tgcagggggt agtgtagccc ggggtgagga
     4681 caaggatggc gtcaagtacc aagtgtcact ctccgagagc atgaagatgg aaggcggcag
     4741 tcccctgcgc ccgggcctag ctaccatggg cacccagcct gcaacacacc acccgatacc
     4801 cacctatccc tcctatgtgc ccgccgcagc tcatgactat ggcagcagtc tcttccatcc
     4861 aggaggcttc ctgggtggcc ccgcctccag cttcacccct aagcagagaa gcaaggctcg
     4921 ctcctgctca gaaggccggg agtgtgtcaa ctgtggtgcc acagccaccc ctctctggcg
     4981 acgagatggc acgggccact acctgtgcaa tgcctgtggg ctctaccaca agatgaatgg
     5041 acagaaccgg ccgctcatca agcccaagcg gaggctgtct gctgccagaa gagcgggcac
     5101 ctgttgtgca aattgtcaga cgacaaccac caccttatgg cgccggaacg ccaacgggga
     5161 ccctgtgtgc aacgcctgtg gcctctacta caagctgcac aatgttaaca ggccactgac
     5221 catgaagaag gaagggatcc agacccggaa tcggaagatg tccagcaaat ccaagaaaag
     5281 caagaaaggg gctgaatgtt tcgaggagct ctccaagtgc atgcaagaga agtcaccgcc
     5341 cttcagtgcg gctgccctgg ctggacacat ggcacctgtg ggacacctcc caccttttag
     5401 tcactctgga cacatcctac ccacgcccac gcctatccac ccttcctcca gtctctcttt
     5461 tggccacccc cacccgtcca gcatggtgac tgccatgggc tagggatcca ttcgatatca
     5521 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     5581 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     5641 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     5701 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     5761 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     5821 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     5881 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     5941 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     6001 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     6061 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     6121 atcgataccg tcgacctcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     6181 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     6241 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     6301 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata
     6361 tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag aactacacac
     6421 cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
     6481 agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga
     6541 gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc
     6601 tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag
     6661 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     6721 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     6781 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc
     6841 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     6901 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     6961 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     7021 agcaaggggg aggattggga agacaatagc aggcatgtga gcaaaaggcc agcaaaaggc
     7081 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7141 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7201 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7261 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7321 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7381 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7441 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7501 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7561 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7621 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7681 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7741 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7801 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7861 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7921 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7981 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8041 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8101 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8161 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8221 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8281 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8341 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8401 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8461 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8521 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8581 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8641 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8701 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8761 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8821 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     8881 agatctcccg atcccc
//
