LOCUS       sequence-382207-        8332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of H2B-mTurquoise in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8332)
  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 8332)
  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-H2B-mTurquoise
FEATURES             Location/Qualifiers
     source          1..8332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..45)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        217..596
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        598..796
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     751..771
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             814..994
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1041..1166
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1659..1892
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2077..2121
                     /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    2419..2536
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2601..3812
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3649..3668
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3833..4210
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      4223..4232
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4229..4231,4232..4234,4235..4948)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2010)"
                     /label=mTurquoise
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4274..4295)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4535..4554)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4882..4903
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4977..5565
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5030..5050)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5448..5459)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5568..5584)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5569..5585)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6090..6270
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6296..6313)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6302..6413
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(6559..7147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6639..6658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7318..8109,8110..8178))
                     /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     7941..7960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8179..8283)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
       61 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
      121 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
      181 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
      241 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      301 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      361 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      421 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      481 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      541 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      601 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      661 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      721 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      781 aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga
      841 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      901 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      961 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
     1021 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
     1081 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
     1141 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
     1201 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
     1261 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     1321 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1381 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1441 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1501 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     1561 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     1621 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     1681 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     1741 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     1801 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     1861 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     1921 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     1981 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     2041 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     2101 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     2161 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     2221 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     2281 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     2341 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
     2401 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     2461 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     2521 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttaattaa
     2581 cccgtgtcgg ctccagatct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc
     2641 cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc
     2701 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     2761 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     2821 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     2881 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     2941 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     3001 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     3061 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     3121 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     3181 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     3241 acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     3301 gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg
     3361 tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga
     3421 gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg
     3481 gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc
     3541 tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga
     3601 ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt
     3661 tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg
     3721 aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gactagtaaa ttgtccgcta
     3781 aattctggcc gtttttggct tttttgttag acgaagcttg ggcccgggat ccatgccaga
     3841 gccagcgaag tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca
     3901 gaagaaaggc ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta
     3961 caaggttctg aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat
     4021 gaattcgttt gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca
     4081 ttacaacaag cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct
     4141 gcctggggag ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac
     4201 cagcgctaag gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     4261 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     4321 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     4381 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgtcctggg gcgtgcagtg
     4441 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     4501 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     4561 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     4621 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacatca gcgacaacgt
     4681 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     4741 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     4801 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga
     4861 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     4921 tctcggcatg gacgagctgt acaagtaagg atccattcga tatcaagctt atcgataatc
     4981 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     5041 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     5101 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     5161 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     5221 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     5281 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     5341 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     5401 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     5461 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     5521 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
     5581 ctcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
     5641 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg
     5701 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     5761 aggggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga
     5821 tctaccacac acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca
     5881 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag
     5941 aagaagccaa tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg
     6001 atgacccgga gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca
     6061 tggcccgaga gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct
     6121 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6181 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6241 ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg
     6301 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     6361 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     6421 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     6481 tgggaagaca atagcaggca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6541 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6601 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6661 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6721 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6781 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6841 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6901 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6961 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     7021 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7081 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7141 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7201 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7261 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7321 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7381 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7441 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7501 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7561 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7621 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7681 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7741 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7801 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7861 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7921 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7981 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8041 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8101 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8161 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8221 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8281 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tc
//
