LOCUS       sequence-382293-        8729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of mCherry fused to human Gata2 with 
            mutation R396Q 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-R396Q
FEATURES             Location/Qualifiers
     source          1..8729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..28
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(43..48,49..66,67..73)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    81..97
                     /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     86..108
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     105..121
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     105..121
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     140..160
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        142..160
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        188..414
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             415..595
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    642..767
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1260..1493
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1678..1722
                     /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    1989..2106
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2140..2546
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             2581..3282
                     /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(2714..2732)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2982..3001
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3176..3199
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    4939..5342
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             5435..5668
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5740..5861
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5777..5796)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5831..5850
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5901..6036
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6056..6075)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6057..6075)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6085..6102)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6085..6101)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6094..6116)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6243..6698
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6330..6349)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6540..6561
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6724..6828
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6829..6897,6898..7689)
                     /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(7047..7066)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7860..8448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8349..8368
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8602..8619
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
       61 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      121 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
      181 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
      241 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      301 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      361 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
      421 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      481 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      541 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
      601 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
      661 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
      721 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
      781 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
      841 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
      901 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
      961 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1021 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1081 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1141 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1201 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1261 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1321 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1381 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1441 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1501 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1561 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     1621 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     1681 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     1741 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     1801 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     1861 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     1921 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     1981 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     2041 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     2101 aattttatcg atcacgagac tagcctcgag ctggctgcag taacgccatt ttgcaaggca
     2161 tggaaaaata ccaaaccaag aatagagaag ttcagatcaa gggcgggtac atgaaaatag
     2221 ctaacgttgg gccaaacagg atatctgcgg tgagcagttt cggccccggc ccggggccaa
     2281 gaacagatgg tcaccgcagt ttcggccccg gcccgaggcc aagaacagat ggtccccaga
     2341 tatggcccaa ccctcagcag tttcttaaga cccatcagat gtttccaggc tcccccaagg
     2401 acctgaaatg accctgcgcc ttatttgaat taaccaatca gcctgcttct cgcttctgtt
     2461 cgcgcgcttc tgcttcccga gctctataaa agagctcaca acccctcact cggcgcgcca
     2521 gtcctccgac agactgagtc ggccggtcga atcaagctta tcgataccgt cgacggatcc
     2581 atggtgagca agggcgagga ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag
     2641 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     2701 ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac
     2761 atcctgtccc cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc
     2821 cccgattaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     2881 gaggacggcg gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc
     2941 tacaaggtga agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag
     3001 accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc
     3061 gagatccacc aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagacc
     3121 atctacatgg ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg
     3181 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc
     3241 cgccaccacc tgttcctgta cggcatggac gagctgtaca aggaggtggc gcccgagcag
     3301 ccgcgctgga tggcgcaccc ggccgtgctg aatgcgcagc accccgactc acaccacccg
     3361 ggcctggcgc acaactacat ggaacccgcg cagctgctgc ctccagacga ggtggacgtc
     3421 ttcttcaatc acctcgactc gcagggcaac ccctactatg ccaaccccgc tcacgcgcgg
     3481 gcgcgcgtct cctacagccc cgcgcacgcc cgcctgaccg gaggccagat gtgccgccca
     3541 cacttgttgc acagcccggg tttgccctgg ctggacgggg gcaaagcagc cctctctgcc
     3601 gctgcggccc accaccacaa cccctggacc gtgagcccct tctccaagac gccactgcac
     3661 ccctcagctg ctggaggccc tggaggccca ctctctgtgt acccaggggc tgggggtggg
     3721 agcgggggag gcagcgggag ctcagtggcc tccctcaccc ctacagcagc ccactctggc
     3781 tcccaccttt tcggcttccc acccacgcca cccaaagaag tgtctcctga ccctagcacc
     3841 acgggggctg cgtctccagc ctcatcttcc gcggggggta gtgcagcccg aggagaggac
     3901 aaggacggcg tcaagtacca ggtgtcactg acggagagca tgaagatgga aagtggcagt
     3961 cccctgcgcc caggcctagc tactatgggc acccagcctg ctacacacca ccccatcccc
     4021 acctacccct cctatgtgcc ggcggctgcc cacgactaca gcagcggact cttccacccc
     4081 ggaggcttcc tggggggacc ggcctccagc ttcaccccta agcagcgcag caaggctcgt
     4141 tcctgttcag aaggccggga gtgtgtcaac tgtggggcca cagccacccc tctctggcgg
     4201 cgggacggca ccggccacta cctgtgcaat gcctgtggcc tctaccacaa gatgaatggg
     4261 cagaaccgac cactcatcaa gcccaagcga agactgtcgg ccgccagaag agccggcacc
     4321 tgttgtgcaa attgtcagac gacaaccacc accttatggc gccgaaacgc caacggggac
     4381 cctgtctgca acgcctgtgg cctctactac aagctgcaca atgttaacag gccactgacc
     4441 atgaagaagg aagggatcca gactcagaac cggaagatgt ccaacaagtc caagaagagc
     4501 aagaaagggg cggagtgctt cgaggagctg tcaaagtgca tgcaggagaa gtcatccccc
     4561 ttcagtgcag ctgccctggc tggacacatg gcacctgtgg gccacctccc gcccttcagc
     4621 cactccggac acatcctgcc cactccgacg cccatccacc cctcctccag cctctccttc
     4681 ggccaccccc acccgtccag catggtgacc gccatgggct agggatcccc cgggctgcag
     4741 gaattcgagc atcttaccgc catttattcc catatttgtt ctgtttttct tgatttgggt
     4801 atacatttaa atgttaataa aacaaaatgg tggggcaatc atttacattt ttagggatat
     4861 gtaattacta gttcaggtgt attgccacaa gacaaacatg ttaagaaact ttcccgttat
     4921 ttacgctctg ttcctgttaa tcaacctctg gattacaaaa tttgtgaaag attgactgat
     4981 attcttaact atgttgctcc ttttacgctg tgtggatatg ctgctttaat gcctctgtat
     5041 catgctattg cttcccgtac ggctttcgtt ttctcctcct tgtataaatc ctggttgctg
     5101 tctctttatg aggagttgtg gcccgttgtc cgtcaacgtg gcgtggtgtg ctctgtgttt
     5161 gctgacgcaa cccccactgg ctggggcatt gccaccacct gtcaactcct ttctgggact
     5221 ttcgctttcc ccctcccgat cgccacggca gaactcatcg ccgcctgcct tgcccgctgc
     5281 tggacagggg ctaggttgct gggcactgat aattccgtgg tgttgtcggg gaagctgacg
     5341 tcctttgcaa ttcgatatca agctgtacct ttaagaccaa tgacttacaa ggcagctgta
     5401 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     5461 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     5521 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     5581 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     5641 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     5701 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     5761 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5821 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     5881 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5941 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     6001 tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta
     6061 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     6121 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     6181 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     6241 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     6301 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6361 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6421 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     6481 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6541 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     6601 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     6661 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     6721 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     6781 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     6841 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     6901 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6961 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     7021 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     7081 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     7141 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     7201 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     7261 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     7321 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     7381 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     7441 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     7501 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     7561 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     7621 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     7681 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     7741 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     7801 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     7861 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     7921 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     7981 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     8041 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     8101 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     8161 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     8221 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     8281 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     8341 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     8401 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     8461 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     8521 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     8581 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     8641 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     8701 cccgactgga aagcgggcag tgagcgcaa
//
