LOCUS       sequence-382210-        8173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression 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-MD-mTurquoise
FEATURES             Location/Qualifiers
     source          1..8173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(37..56)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        228..607
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        609..807
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     762..782
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             825..1005
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1052..1177
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1670..1903
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2088..2132
                     /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    2430..2547
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2612..3823
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3660..3679
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(4084..4086,4087..4800)
                     /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(4126..4147)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4387..4406)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4734..4755
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4829..5417
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4882..4902)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5300..5311)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5420..5436)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5421..5437)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5942..6122
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6148..6165)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6154..6265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(6411..6999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6491..6510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7170..7961,7962..8030))
                     /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     7793..7812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8031..8135)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
       61 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
      121 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
      181 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
      241 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      301 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      361 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      421 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      481 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      541 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      601 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      661 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      721 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      781 tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct ctctggttag
      841 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
      901 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
      961 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     1021 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     1081 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     1141 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     1201 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     1261 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     1321 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     1381 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     1441 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     1501 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     1561 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     1621 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     1681 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     1741 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     1801 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     1861 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     1921 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     1981 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     2041 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     2101 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     2161 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     2221 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     2281 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag
     2341 agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt gcgccaattc
     2401 tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac
     2461 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     2521 aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag agatccagtt
     2581 tggttaatta acccgtgtcg gctccagatc tggcctccgc gccgggtttt ggcgcctccc
     2641 gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt
     2701 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     2761 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     2821 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     2881 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     2941 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     3001 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     3061 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     3121 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     3181 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     3241 gggcggcaag aacccaaggt cttgaggcct tcgctaatgc gggaaagctc ttattcgggt
     3301 gagatgggct ggggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac
     3361 tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg
     3421 tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa
     3481 tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca
     3541 gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga
     3601 gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct
     3661 gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta
     3721 ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agactagtaa
     3781 attgtccgct aaattctggc cgtttttggc ttttttgtta gacgaagctt gggcccggga
     3841 tccatggctg agaacaaggc aaaagtgagc atggcaggag ccaagagggt gcccgtcacc
     3901 gtgacagccg cttccaaacc aggcctgagg cccagaacag cactcggtga catcggcaat
     3961 aaggtctctg aggaactgca ggcccgggtg cctctcaagc gagaagctaa aactctgggc
     4021 accggaaagg ggaccgtcaa agctctcccc aagcctgtgg agaaagtccc tgtgtgcgaa
     4081 ccagtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     4141 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     4201 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     4261 ctcgtgacca ccctgtcctg gggcgtgcag tgcttcgccc gctaccccga ccacatgaag
     4321 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     4381 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     4441 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     4501 aagctggagt acaactacat cagcgacaac gtctatatca ccgccgacaa gcagaagaac
     4561 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     4621 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     4681 tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     4741 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     4801 ggatccattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     4861 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     4921 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     4981 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5041 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5101 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5161 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     5221 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5281 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     5341 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5401 ttgggccgcc tccccgcatc gataccgtcg acctcgagac ctagaaaaac atggagcaat
     5461 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     5521 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     5581 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     5641 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     5701 ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta
     5761 caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg
     5821 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg
     5881 gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc cggactgtac
     5941 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     6001 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6061 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6121 cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     6181 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     6241 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     6301 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgtgagca
     6361 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6421 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6481 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6541 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6601 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6661 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6721 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6781 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6841 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6901 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6961 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7021 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7081 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7141 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7201 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7261 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7321 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7381 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7441 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7501 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7561 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7621 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7681 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7741 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7801 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7861 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7921 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7981 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8041 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8101 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8161 gacgtcgacg gat
//
