LOCUS       sequence-380076-        8729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of mCherry fused to human Gata2 with 
            mutation C373R 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-C373R
FEATURES             Location/Qualifiers
     source          1..8729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    57..78
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(93..98,99..116,117..123)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    131..147
                     /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     136..158
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     155..171
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     155..171
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     190..210
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        192..210
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        238..464
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             465..645
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    692..817
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1310..1543
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1728..1772
                     /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    2039..2156
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2190..2596
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             2631..3332
                     /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(2764..2782)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3032..3051
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3226..3249
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    4989..5392
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             5485..5718
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5790..5911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5827..5846)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5881..5900
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5951..6086
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6106..6125)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6107..6125)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6135..6152)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6135..6151)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6144..6166)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6293..6748
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6380..6399)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6590..6611
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6774..6878
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6879..6947,6948..7739)
                     /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(7097..7116)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7910..8498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8399..8418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8652..8669
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
       61 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
      121 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
      181 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt
      241 agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct
      301 tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg
      361 ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc
      421 attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag
      481 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
      541 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
      601 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
      661 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
      721 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
      781 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
      841 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
      901 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
      961 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     1021 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     1081 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     1141 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     1201 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     1261 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     1321 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     1381 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     1441 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     1501 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     1561 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     1621 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     1681 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     1741 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     1801 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     1861 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     1921 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag
     1981 agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat cggttaactt
     2041 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     2101 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttatcg
     2161 atcacgagac tagcctcgag ctggctgcag taacgccatt ttgcaaggca tggaaaaata
     2221 ccaaaccaag aatagagaag ttcagatcaa gggcgggtac atgaaaatag ctaacgttgg
     2281 gccaaacagg atatctgcgg tgagcagttt cggccccggc ccggggccaa gaacagatgg
     2341 tcaccgcagt ttcggccccg gcccgaggcc aagaacagat ggtccccaga tatggcccaa
     2401 ccctcagcag tttcttaaga cccatcagat gtttccaggc tcccccaagg acctgaaatg
     2461 accctgcgcc ttatttgaat taaccaatca gcctgcttct cgcttctgtt cgcgcgcttc
     2521 tgcttcccga gctctataaa agagctcaca acccctcact cggcgcgcca gtcctccgac
     2581 agactgagtc ggccggtcga atcaagctta tcgataccgt cgacggatcc atggtgagca
     2641 agggcgagga ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga
     2701 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     2761 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
     2821 cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca
     2881 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     2941 gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga
     3001 agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct
     3061 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc
     3121 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg
     3181 ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct
     3241 cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc
     3301 tgttcctgta cggcatggac gagctgtaca aggaggtggc gcccgagcag ccgcgctgga
     3361 tggcgcaccc ggccgtgctg aatgcgcagc accccgactc acaccacccg ggcctggcgc
     3421 acaactacat ggaacccgcg cagctgctgc ctccagacga ggtggacgtc ttcttcaatc
     3481 acctcgactc gcagggcaac ccctactatg ccaaccccgc tcacgcgcgg gcgcgcgtct
     3541 cctacagccc cgcgcacgcc cgcctgaccg gaggccagat gtgccgccca cacttgttgc
     3601 acagcccggg tttgccctgg ctggacgggg gcaaagcagc cctctctgcc gctgcggccc
     3661 accaccacaa cccctggacc gtgagcccct tctccaagac gccactgcac ccctcagctg
     3721 ctggaggccc tggaggccca ctctctgtgt acccaggggc tgggggtggg agcgggggag
     3781 gcagcgggag ctcagtggcc tccctcaccc ctacagcagc ccactctggc tcccaccttt
     3841 tcggcttccc acccacgcca cccaaagaag tgtctcctga ccctagcacc acgggggctg
     3901 cgtctccagc ctcatcttcc gcggggggta gtgcagcccg aggagaggac aaggacggcg
     3961 tcaagtacca ggtgtcactg acggagagca tgaagatgga aagtggcagt cccctgcgcc
     4021 caggcctagc tactatgggc acccagcctg ctacacacca ccccatcccc acctacccct
     4081 cctatgtgcc ggcggctgcc cacgactaca gcagcggact cttccacccc ggaggcttcc
     4141 tggggggacc ggcctccagc ttcaccccta agcagcgcag caaggctcgt tcctgttcag
     4201 aaggccggga gtgtgtcaac tgtggggcca cagccacccc tctctggcgg cgggacggca
     4261 ccggccacta cctgtgcaat gcctgtggcc tctaccacaa gatgaatggg cagaaccgac
     4321 cactcatcaa gcccaagcga agactgtcgg ccgccagaag agccggcacc tgttgtgcaa
     4381 attgtcagac gacaaccacc accttatggc gccgaaacgc caacggggac cctgtctgca
     4441 acgcccgtgg cctctactac aagctgcaca atgttaacag gccactgacc atgaagaagg
     4501 aagggatcca gactcggaac cggaagatgt ccaacaagtc caagaagagc aagaaagggg
     4561 cggagtgctt cgaggagctg tcaaagtgca tgcaggagaa gtcatccccc ttcagtgcag
     4621 ctgccctggc tggacacatg gcacctgtgg gccacctccc gcccttcagc cactccggac
     4681 acatcctgcc cactccgacg cccatccacc cctcctccag cctctccttc ggccaccccc
     4741 acccgtccag catggtgacc gccatgggct agggatcccc cgggctgcag gaattcgagc
     4801 atcttaccgc catttattcc catatttgtt ctgtttttct tgatttgggt atacatttaa
     4861 atgttaataa aacaaaatgg tggggcaatc atttacattt ttagggatat gtaattacta
     4921 gttcaggtgt attgccacaa gacaaacatg ttaagaaact ttcccgttat ttacgctctg
     4981 ttcctgttaa tcaacctctg gattacaaaa tttgtgaaag attgactgat attcttaact
     5041 atgttgctcc ttttacgctg tgtggatatg ctgctttaat gcctctgtat catgctattg
     5101 cttcccgtac ggctttcgtt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     5161 aggagttgtg gcccgttgtc cgtcaacgtg gcgtggtgtg ctctgtgttt gctgacgcaa
     5221 cccccactgg ctggggcatt gccaccacct gtcaactcct ttctgggact ttcgctttcc
     5281 ccctcccgat cgccacggca gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     5341 ctaggttgct gggcactgat aattccgtgg tgttgtcggg gaagctgacg tcctttgcaa
     5401 ttcgatatca agctgtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5461 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     5521 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5581 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5641 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     5701 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     5761 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     5821 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5881 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     5941 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6001 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6061 gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg
     6121 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     6181 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     6241 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     6301 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6361 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6421 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6481 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     6541 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     6601 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     6661 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6721 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     6781 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     6841 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6901 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6961 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     7021 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     7081 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     7141 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     7201 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     7261 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     7321 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     7381 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     7441 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     7501 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     7561 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     7621 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     7681 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     7741 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     7801 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7861 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7921 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7981 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     8041 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     8101 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     8161 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     8221 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     8281 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     8341 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     8401 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     8461 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     8521 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     8581 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     8641 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     8701 ctctccccgc gcgttggccg attcattaa
//
