LOCUS       sequence-380075-        8729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of mCherry fused to human Gata2 with 
            mutation L359V in mammalian cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8729)
  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 8729)
  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: SFFV-mCh-GATA2-L359V
FEATURES             Location/Qualifiers
     source          1..8729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    89..110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(125..130,131..148,149..155)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    163..179
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     168..190
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     187..203
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     187..203
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     222..242
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        224..242
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        270..496
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             497..677
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    724..849
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1342..1575
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1760..1804
                     /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    2071..2188
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2222..2628
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             2663..3364
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(2796..2814)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3064..3083
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3258..3281
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    5021..5424
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             5517..5750
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5822..5943
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5859..5878)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5913..5932
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5983..6118
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6138..6157)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6139..6157)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6167..6184)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6167..6183)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6176..6198)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6325..6780
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6412..6431)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6622..6643
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6806..6910
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6911..6979,6980..7771)
                     /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     complement(7129..7148)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7942..8530
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8431..8450
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8684..8701
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
       61 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
      121 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
      181 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
      241 gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg
      301 caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg
      361 ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct
      421 gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct
      481 agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
      541 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
      601 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
      661 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
      721 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
      781 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
      841 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
      901 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
      961 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     1021 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     1081 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     1141 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     1201 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     1261 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     1321 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     1381 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     1441 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     1501 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     1561 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     1621 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     1681 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     1741 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     1801 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     1861 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     1921 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     1981 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     2041 agtgaacgga tctcgacggt atcggttaac ttttaaaaga aaagggggga ttggggggta
     2101 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     2161 aaaacaaatt acaaaaattc aaaattttat cgatcacgag actagcctcg agctggctgc
     2221 agtaacgcca ttttgcaagg catggaaaaa taccaaacca agaatagaga agttcagatc
     2281 aagggcgggt acatgaaaat agctaacgtt gggccaaaca ggatatctgc ggtgagcagt
     2341 ttcggccccg gcccggggcc aagaacagat ggtcaccgca gtttcggccc cggcccgagg
     2401 ccaagaacag atggtcccca gatatggccc aaccctcagc agtttcttaa gacccatcag
     2461 atgtttccag gctcccccaa ggacctgaaa tgaccctgcg ccttatttga attaaccaat
     2521 cagcctgctt ctcgcttctg ttcgcgcgct tctgcttccc gagctctata aaagagctca
     2581 caacccctca ctcggcgcgc cagtcctccg acagactgag tcggccggtc gaatcaagct
     2641 tatcgatacc gtcgacggat ccatggtgag caagggcgag gaggtcatca aagagttcat
     2701 gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga
     2761 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg
     2821 ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta
     2881 cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa
     2941 gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc
     3001 cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga
     3061 cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc
     3121 ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca
     3181 ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta
     3241 ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     3301 acagtacgag cgctccgagg gccgccacca cctgttcctg tacggcatgg acgagctgta
     3361 caaggaggtg gcgcccgagc agccgcgctg gatggcgcac ccggccgtgc tgaatgcgca
     3421 gcaccccgac tcacaccacc cgggcctggc gcacaactac atggaacccg cgcagctgct
     3481 gcctccagac gaggtggacg tcttcttcaa tcacctcgac tcgcagggca acccctacta
     3541 tgccaacccc gctcacgcgc gggcgcgcgt ctcctacagc cccgcgcacg cccgcctgac
     3601 cggaggccag atgtgccgcc cacacttgtt gcacagcccg ggtttgccct ggctggacgg
     3661 gggcaaagca gccctctctg ccgctgcggc ccaccaccac aacccctgga ccgtgagccc
     3721 cttctccaag acgccactgc acccctcagc tgctggaggc cctggaggcc cactctctgt
     3781 gtacccaggg gctgggggtg ggagcggggg aggcagcggg agctcagtgg cctccctcac
     3841 ccctacagca gcccactctg gctcccacct tttcggcttc ccacccacgc cacccaaaga
     3901 agtgtctcct gaccctagca ccacgggggc tgcgtctcca gcctcatctt ccgcgggggg
     3961 tagtgcagcc cgaggagagg acaaggacgg cgtcaagtac caggtgtcac tgacggagag
     4021 catgaagatg gaaagtggca gtcccctgcg cccaggccta gctactatgg gcacccagcc
     4081 tgctacacac caccccatcc ccacctaccc ctcctatgtg ccggcggctg cccacgacta
     4141 cagcagcgga ctcttccacc ccggaggctt cctgggggga ccggcctcca gcttcacccc
     4201 taagcagcgc agcaaggctc gttcctgttc agaaggccgg gagtgtgtca actgtggggc
     4261 cacagccacc cctctctggc ggcgggacgg caccggccac tacctgtgca atgcctgtgg
     4321 cctctaccac aagatgaatg ggcagaaccg accactcatc aagcccaagc gaagactgtc
     4381 ggccgccaga agagccggca cctgttgtgc aaattgtcag acgacaacca ccaccgtatg
     4441 gcgccgaaac gccaacgggg accctgtctg caacgcctgt ggcctctact acaagctgca
     4501 caatgttaac aggccactga ccatgaagaa ggaagggatc cagactcgga accggaagat
     4561 gtccaacaag tccaagaaga gcaagaaagg ggcggagtgc ttcgaggagc tgtcaaagtg
     4621 catgcaggag aagtcatccc ccttcagtgc agctgccctg gctggacaca tggcacctgt
     4681 gggccacctc ccgcccttca gccactccgg acacatcctg cccactccga cgcccatcca
     4741 cccctcctcc agcctctcct tcggccaccc ccacccgtcc agcatggtga ccgccatggg
     4801 ctagggatcc cccgggctgc aggaattcga gcatcttacc gccatttatt cccatatttg
     4861 ttctgttttt cttgatttgg gtatacattt aaatgttaat aaaacaaaat ggtggggcaa
     4921 tcatttacat ttttagggat atgtaattac tagttcaggt gtattgccac aagacaaaca
     4981 tgttaagaaa ctttcccgtt atttacgctc tgttcctgtt aatcaacctc tggattacaa
     5041 aatttgtgaa agattgactg atattcttaa ctatgttgct ccttttacgc tgtgtggata
     5101 tgctgcttta atgcctctgt atcatgctat tgcttcccgt acggctttcg ttttctcctc
     5161 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tccgtcaacg
     5221 tggcgtggtg tgctctgtgt ttgctgacgc aacccccact ggctggggca ttgccaccac
     5281 ctgtcaactc ctttctggga ctttcgcttt ccccctcccg atcgccacgg cagaactcat
     5341 cgccgcctgc cttgcccgct gctggacagg ggctaggttg ctgggcactg ataattccgt
     5401 ggtgttgtcg gggaagctga cgtcctttgc aattcgatat caagctgtac ctttaagacc
     5461 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     5521 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     5581 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     5641 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     5701 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     5761 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     5821 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5881 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5941 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg
     6001 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6061 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6121 gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     6181 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     6241 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     6301 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     6361 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     6421 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     6481 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     6541 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     6601 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6661 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6721 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     6781 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6841 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6901 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6961 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     7021 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     7081 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     7141 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     7201 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     7261 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     7321 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     7381 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     7441 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     7501 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     7561 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     7621 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     7681 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     7741 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     7801 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7861 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7921 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7981 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     8041 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     8101 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     8161 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     8221 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     8281 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     8341 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     8401 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     8461 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     8521 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     8581 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     8641 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     8701 ggaagcggaa gagcgcccaa tacgcaaac
//
