LOCUS       sequence-382235-        8896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression 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-MD-Gata2
FEATURES             Location/Qualifiers
     source          1..8896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(14..33)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        205..584
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        586..784
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     739..759
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             802..982
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1029..1154
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1647..1880
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2065..2109
                     /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    2407..2524
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2589..3800
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3637..3656
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     misc_feature    5529..6117
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5582..5602)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6000..6011)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6120..6136)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6121..6137)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6642..6822
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6848..6865)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6854..6965
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(7111..7699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7191..7210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7870..8661,8662..8730))
                     /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     8493..8512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8731..8835)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
       61 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
      121 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
      181 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
      241 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      301 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      361 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      421 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      481 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      541 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      601 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      661 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      721 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      781 gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
      841 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
      901 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
      961 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     1021 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     1081 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     1141 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     1201 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     1261 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     1321 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     1381 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     1441 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     1501 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     1561 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     1621 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     1681 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     1741 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     1801 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     1861 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     1921 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     1981 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     2041 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     2101 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     2161 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     2221 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     2281 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
     2341 cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat
     2401 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     2461 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     2521 ttttcgggtt tattacaggg acagcagaga tccagtttgg ttaattaacc cgtgtcggct
     2581 ccagatctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc
     2641 gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct
     2701 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     2761 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     2821 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     2881 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     2941 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     3001 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     3061 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     3121 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     3181 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     3241 gaggccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg gcaccatctg
     3301 gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg
     3361 gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc
     3421 gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg
     3481 taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc
     3541 tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc
     3601 tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc
     3661 gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat
     3721 ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt gtccgctaaa ttctggccgt
     3781 ttttggcttt tttgttagac gaagcttggg cccgggatcc atggctgaga acaaggcaaa
     3841 agtgagcatg gcaggagcca agagggtgcc cgtcaccgtg acagccgctt ccaaaccagg
     3901 cctgaggccc agaacagcac tcggtgacat cggcaataag gtctctgagg aactgcaggc
     3961 ccgggtgcct ctcaagcgag aagctaaaac tctgggcacc ggaaagggga ccgtcaaagc
     4021 tctccccaag cctgtggaga aagtccctgt gtgcgaacca gaggtggcgc ctgagcagcc
     4081 tcgctggatg gcgcaccccg ccgtattgaa tgcgcagcac cccgactcgc accatccggg
     4141 cctggcgcat aactacatgg agccagcaca gctgctgcct cccgacgagg tggatgtctt
     4201 cttcaaccat ctcgactcgc agggcaaccc ttactacgcc aacccggccc acgcgcgcgc
     4261 gcgcgtttcc tacagcccgg cgcatgcccg tctcaccgga ggccagatgt gccgaccaca
     4321 cttgttgcac agcccaggct tgccgtggct ggacgggggc aaagcagctc tctctgccgc
     4381 cgctgcccat caccacagtc cctggaccgt cagcccgttc tccaagaccc cgctgcaccc
     4441 ctcagctgct ggagcacccg gagggcctct gtctgtttac ccaggggctg cgggtgggag
     4501 cgggggaggc agtgggagct ccgtggcctc cctcaccccc actgcagccc actcgggctc
     4561 ccatctcttc ggcttcccac ccacgccacc caaagaagtg tctccagacc ccagcacaac
     4621 aggagctgct tccccggcct cttcttctgc agggggtagt gtagcccggg gtgaggacaa
     4681 ggatggcgtc aagtaccaag tgtcactctc cgagagcatg aagatggaag gcggcagtcc
     4741 cctgcgcccg ggcctagcta ccatgggcac ccagcctgca acacaccacc cgatacccac
     4801 ctatccctcc tatgtgcccg ccgcagctca tgactatggc agcagtctct tccatccagg
     4861 aggcttcctg ggtggccccg cctccagctt cacccctaag cagagaagca aggctcgctc
     4921 ctgctcagaa ggccgggagt gtgtcaactg tggtgccaca gccacccctc tctggcgacg
     4981 agatggcacg ggccactacc tgtgcaatgc ctgtgggctc taccacaaga tgaatggaca
     5041 gaaccggccg ctcatcaagc ccaagcggag gctgtctgct gccagaagag cgggcacctg
     5101 ttgtgcaaat tgtcagacga caaccaccac cttatggcgc cggaacgcca acggggaccc
     5161 tgtgtgcaac gcctgtggcc tctactacaa gctgcacaat gttaacaggc cactgaccat
     5221 gaagaaggaa gggatccaga cccggaatcg gaagatgtcc agcaaatcca agaaaagcaa
     5281 gaaaggggct gaatgtttcg aggagctctc caagtgcatg caagagaagt caccgccctt
     5341 cagtgcggct gccctggctg gacacatggc acctgtggga cacctcccac cttttagtca
     5401 ctctggacac atcctaccca cgcccacgcc tatccaccct tcctccagtc tctcttttgg
     5461 ccacccccac ccgtccagca tggtgactgc catgggctag ggatccattc gatatcaagc
     5521 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     5581 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     5641 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     5701 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     5761 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     5821 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     5881 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     5941 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     6001 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     6061 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     6121 gataccgtcg acctcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag
     6181 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag
     6241 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     6301 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc
     6361 ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag
     6421 ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
     6481 aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc
     6541 tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag
     6601 catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc
     6661 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6721 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6781 gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc
     6841 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     6901 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     6961 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     7021 aagggggagg attgggaaga caatagcagg catgtgagca aaaggccagc aaaaggccag
     7081 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7141 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7201 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7261 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7321 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7381 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7441 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7501 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7561 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7621 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7681 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7741 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7801 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7861 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7921 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7981 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     8041 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8101 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8161 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8221 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8281 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8341 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8401 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8461 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8521 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8581 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8641 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8701 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8761 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8821 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     8881 tctcccgatc ccctat
//
