LOCUS       sequence-382209-        8173 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
            mTurquoise in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8173)
  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 8173)
  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-mTurquoise
FEATURES             Location/Qualifiers
     source          1..8173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        247..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        628..826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     781..801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             844..1024
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1071..1196
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1689..1922
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2107..2151
                     /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    2449..2566
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2631..3842
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3679..3698
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(4103..4105,4106..4819)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2010)"
                     /label=mTurquoise
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4145..4166)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4406..4425)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4753..4774
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4848..5436
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4901..4921)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5319..5330)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5439..5455)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5440..5456)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5961..6141
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6167..6184)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6173..6284
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(6430..7018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6510..6529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7189..7980,7981..8049))
                     /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     7812..7831
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8050..8154)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     1021 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     1081 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1141 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1201 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1261 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1321 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1381 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1441 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1501 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     1561 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     1621 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     1681 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     1741 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     1801 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     1861 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     1921 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     1981 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     2041 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     2101 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     2161 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     2221 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     2281 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     2341 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg
     2401 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa
     2461 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     2521 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     2581 ggacagcaga gatccagttt ggttaattaa cccgtgtcgg ctccagatct ggcctccgcg
     2641 ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga
     2701 cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
     2761 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
     2821 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
     2881 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     2941 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     3001 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
     3061 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
     3121 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
     3181 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
     3241 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg
     3301 ggaaagctct tattcgggtg agatgggctg gggcaccatc tggggaccct gacgtgaagt
     3361 ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg
     3421 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
     3481 ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt
     3541 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
     3601 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
     3661 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
     3721 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
     3781 ttcagtgtta gactagtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
     3841 acgaagcttg ggcccgggat ccatggctga gaacaaggca aaagtgagca tggcaggagc
     3901 caagagggtg cccgtcaccg tgacagccgc ttccaaacca ggcctgaggc ccgcaacagc
     3961 actcggtgac atcggcaata aggtctctga ggaactgcag gcccgggtgc ctctcaagcg
     4021 agaagctaaa actctgggca ccggaaaggg gaccgtcaaa gctctcccca agcctgtgga
     4081 gaaagtccct gtgtgcgaac cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     4141 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     4201 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     4261 gcccgtgccc tggcccaccc tcgtgaccac cctgtcctgg ggcgtgcagt gcttcgcccg
     4321 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4381 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4441 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4501 cggcaacatc ctggggcaca agctggagta caactacatc agcgacaacg tctatatcac
     4561 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     4621 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4681 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga
     4741 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4801 ggacgagctg tacaagtaag gatccattcg atatcaagct tatcgataat caacctctgg
     4861 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     4921 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     4981 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5041 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     5101 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5161 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     5221 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     5281 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     5341 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     5401 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgagacc
     5461 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
     5521 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa
     5581 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     5641 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca
     5701 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac
     5761 tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca
     5821 atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
     5881 agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag
     5941 agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     6001 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6061 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6121 cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct
     6181 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     6241 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     6301 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     6361 aatagcaggc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6421 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6481 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6541 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6601 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6661 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6721 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6781 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6841 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6901 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6961 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7021 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7081 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7141 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7201 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7261 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7321 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7381 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7441 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7501 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7561 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7621 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7681 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7741 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7801 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7861 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7921 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7981 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8041 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8101 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8161 tccccgaaaa gtg
//
