LOCUS       sequence-382427-        8729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of mCherry fused to human Gata2 with 
            mutation P304H 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-P304H
FEATURES             Location/Qualifiers
     source          1..8729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(139..158)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(140..158)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(168..185)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(168..184)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(177..199)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      326..781
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(413..432)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     623..644
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        807..911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(912..980,981..1772)
                     /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(1130..1149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1943..2531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2432..2451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2685..2702
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2819..2840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2855..2860,2861..2878,2879..2885)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2893..2909
                     /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     2898..2920
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2917..2933
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2917..2933
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2952..2972
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2954..2972
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3000..3226
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3227..3407
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3454..3579
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4072..4305
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4490..4534
                     /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    4801..4918
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4952..5358
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             5393..6094
                     /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(5526..5544)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5794..5813
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5988..6011
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    7751..8154
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             8247..8480
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8552..8673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8589..8608)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8643..8662
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      join(8713..8729,1..119)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
       61 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
      121 agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
      181 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
      241 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
      301 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
      361 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      421 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      481 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
      541 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
      601 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
      661 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
      721 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
      781 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
      841 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      901 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      961 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1021 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1081 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1141 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1201 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1261 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1321 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1381 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1441 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1501 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1561 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1621 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     1681 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     1741 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1801 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1861 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1921 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1981 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2041 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2101 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2161 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2221 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2281 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2341 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2401 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2461 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2521 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2581 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     2641 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2701 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     2761 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2821 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2881 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     2941 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     3001 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     3061 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     3121 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
     3181 gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt
     3241 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     3301 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     3361 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     3421 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     3481 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     3541 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     3601 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     3661 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     3721 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     3781 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     3841 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     3901 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     3961 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     4021 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     4081 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     4141 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     4201 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     4261 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     4321 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     4381 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     4441 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     4501 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     4561 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     4621 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     4681 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
     4741 agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt atcggttaac
     4801 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
     4861 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttat
     4921 cgatcacgag actagcctcg agctggctgc agtaacgcca ttttgcaagg catggaaaaa
     4981 taccaaacca agaatagaga agttcagatc aagggcgggt acatgaaaat agctaacgtt
     5041 gggccaaaca ggatatctgc ggtgagcagt ttcggccccg gcccggggcc aagaacagat
     5101 ggtcaccgca gtttcggccc cggcccgagg ccaagaacag atggtcccca gatatggccc
     5161 aaccctcagc agtttcttaa gacccatcag atgtttccag gctcccccaa ggacctgaaa
     5221 tgaccctgcg ccttatttga attaaccaat cagcctgctt ctcgcttctg ttcgcgcgct
     5281 tctgcttccc gagctctata aaagagctca caacccctca ctcggcgcgc cagtcctccg
     5341 acagactgag tcggccggtc gaatcaagct tatcgatacc gtcgacggat ccatggtgag
     5401 caagggcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat
     5461 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     5521 gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
     5581 cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta
     5641 caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     5701 cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt
     5761 gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg
     5821 ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca
     5881 ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat
     5941 ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac
     6001 ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca
     6061 cctgttcctg tacggcatgg acgagctgta caaggaggtg gcgcccgagc agccgcgctg
     6121 gatggcgcac ccggccgtgc tgaatgcgca gcaccccgac tcacaccacc cgggcctggc
     6181 gcacaactac atggaacccg cgcagctgct gcctccagac gaggtggacg tcttcttcaa
     6241 tcacctcgac tcgcagggca acccctacta tgccaacccc gctcacgcgc gggcgcgcgt
     6301 ctcctacagc cccgcgcacg cccgcctgac cggaggccag atgtgccgcc cacacttgtt
     6361 gcacagcccg ggtttgccct ggctggacgg gggcaaagca gccctctctg ccgctgcggc
     6421 ccaccaccac aacccctgga ccgtgagccc cttctccaag acgccactgc acccctcagc
     6481 tgctggaggc cctggaggcc cactctctgt gtacccaggg gctgggggtg ggagcggggg
     6541 aggcagcggg agctcagtgg cctccctcac ccctacagca gcccactctg gctcccacct
     6601 tttcggcttc ccacccacgc cacccaaaga agtgtctcct gaccctagca ccacgggggc
     6661 tgcgtctcca gcctcatctt ccgcgggggg tagtgcagcc cgaggagagg acaaggacgg
     6721 cgtcaagtac caggtgtcac tgacggagag catgaagatg gaaagtggca gtcccctgcg
     6781 cccaggccta gctactatgg gcacccagcc tgctacacac caccccatcc ccacctaccc
     6841 ctcctatgtg ccggcggctg cccacgacta cagcagcgga ctcttccacc ccggaggctt
     6901 cctgggggga ccggcctcca gcttcacccc taagcagcgc agcaaggctc gttcctgttc
     6961 agaaggccgg gagtgtgtca actgtggggc cacagccacc catctctggc ggcgggacgg
     7021 caccggccac tacctgtgca atgcctgtgg cctctaccac aagatgaatg ggcagaaccg
     7081 accactcatc aagcccaagc gaagactgtc ggccgccaga agagccggca cctgttgtgc
     7141 aaattgtcag acgacaacca ccaccttatg gcgccgaaac gccaacgggg accctgtctg
     7201 caacgcctgt ggcctctact acaagctgca caatgttaac aggccactga ccatgaagaa
     7261 ggaagggatc cagactcgga accggaagat gtccaacaag tccaagaaga gcaagaaagg
     7321 ggcggagtgc ttcgaggagc tgtcaaagtg catgcaggag aagtcatccc ccttcagtgc
     7381 agctgccctg gctggacaca tggcacctgt gggccacctc ccgcccttca gccactccgg
     7441 acacatcctg cccactccga cgcccatcca cccctcctcc agcctctcct tcggccaccc
     7501 ccacccgtcc agcatggtga ccgccatggg ctagggatcc cccgggctgc aggaattcga
     7561 gcatcttacc gccatttatt cccatatttg ttctgttttt cttgatttgg gtatacattt
     7621 aaatgttaat aaaacaaaat ggtggggcaa tcatttacat ttttagggat atgtaattac
     7681 tagttcaggt gtattgccac aagacaaaca tgttaagaaa ctttcccgtt atttacgctc
     7741 tgttcctgtt aatcaacctc tggattacaa aatttgtgaa agattgactg atattcttaa
     7801 ctatgttgct ccttttacgc tgtgtggata tgctgcttta atgcctctgt atcatgctat
     7861 tgcttcccgt acggctttcg ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     7921 tgaggagttg tggcccgttg tccgtcaacg tggcgtggtg tgctctgtgt ttgctgacgc
     7981 aacccccact ggctggggca ttgccaccac ctgtcaactc ctttctggga ctttcgcttt
     8041 ccccctcccg atcgccacgg cagaactcat cgccgcctgc cttgcccgct gctggacagg
     8101 ggctaggttg ctgggcactg ataattccgt ggtgttgtcg gggaagctga cgtcctttgc
     8161 aattcgatat caagctgtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     8221 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     8281 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     8341 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     8401 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     8461 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     8521 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     8581 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     8641 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     8701 ctaactccgc ccatcccgcc cctaactcc
//
