LOCUS       sequence-396565-        8659 bp    DNA     circular SYN 15-APR-2025
DEFINITION  constitutive expression of mouse Gata2 with mutation L359V in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8659)
  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 8659)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Gata2-L359V
FEATURES             Location/Qualifiers
     source          1..8659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        408..606
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             624..804
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    851..976
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1469..1702
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1887..1931
                     /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    2229..2346
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2411..3622
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3459..3478
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     misc_feature    5114..5702
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5167..5187)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5585..5596)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5705..5721)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5706..5722)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6227..6407
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6433..6450)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6439..6550
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(6696..7284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6776..6795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7455..8246,8247..8315))
                     /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     8078..8097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8316..8420)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8495..8514)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
      661 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
      721 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
      781 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
      841 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
      901 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
      961 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
     1021 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
     1081 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
     1141 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
     1201 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
     1261 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
     1321 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     1381 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     1441 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     1501 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     1561 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     1621 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     1681 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     1741 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     1801 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     1861 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     1921 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     1981 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     2041 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     2101 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     2161 ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc
     2221 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     2281 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     2341 aattttcggg tttattacag ggacagcaga gatccagttt ggttaattaa cccgtgtcgg
     2401 ctccagatct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg
     2461 gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg
     2521 ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag
     2581 gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga
     2641 acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt
     2701 gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg
     2761 ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg
     2821 ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg
     2881 gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt
     2941 ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg
     3001 cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc
     3061 ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg gggcaccatc
     3121 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     3181 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     3241 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc
     3301 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     3361 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     3421 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     3481 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     3541 atttgggtca atatgtaatt ttcagtgtta gactagtaaa ttgtccgcta aattctggcc
     3601 gtttttggct tttttgttag acgaagcttg ggcccgggat ccatggaggt ggcgcctgag
     3661 cagcctcgct ggatggcgca ccccgccgta ttgaatgcgc agcaccccga ctcgcaccat
     3721 ccgggcctgg cgcataacta catggagcca gcacagctgc tgcctcccga cgaggtggat
     3781 gtcttcttca accatctcga ctcgcagggc aacccttact acgccaaccc ggcccacgcg
     3841 cgcgcgcgcg tttcctacag cccggcgcat gcccgtctca ccggaggcca gatgtgccga
     3901 ccacacttgt tgcacagccc aggcttgccg tggctggacg ggggcaaagc agctctctct
     3961 gccgccgctg cccatcacca cagtccctgg accgtcagcc cgttctccaa gaccccgctg
     4021 cacccctcag ctgctggagc acccggaggg cctctgtctg tttacccagg ggctgcgggt
     4081 gggagcgggg gaggcagtgg gagctccgtg gcctccctca cccccactgc agcccactcg
     4141 ggctcccatc tcttcggctt cccacccacg ccacccaaag aagtgtctcc agaccccagc
     4201 acaacaggag ctgcttcccc ggcctcttct tctgcagggg gtagtgtagc ccggggtgag
     4261 gacaaggatg gcgtcaagta ccaagtgtca ctctccgaga gcatgaagat ggaaggcggc
     4321 agtcccctgc gcccgggcct agctaccatg ggcacccagc ctgcaacaca ccacccgata
     4381 cccacctatc cctcctatgt gcccgccgca gctcatgact atggcagcag tctcttccat
     4441 ccaggaggct tcctgggtgg ccccgcctcc agcttcaccc ctaagcagag aagcaaggct
     4501 cgctcctgct cagaaggccg ggagtgtgtc aactgtggtg ccacagccac ccctctctgg
     4561 cgacgagatg gcacgggcca ctacctgtgc aatgcctgtg ggctctacca caagatgaat
     4621 ggacagaacc ggccgctcat caagcccaag cggaggctgt ctgctgccag aagagcgggc
     4681 acctgttgtg caaattgtca gacgacaacc accaccgtat ggcgccggaa cgccaacggg
     4741 gaccctgtgt gcaacgcctg tggcctctac tacaagctgc acaatgttaa caggccactg
     4801 accatgaaga aggaagggat ccagacccgg aatcggaaga tgtccagcaa atccaagaaa
     4861 agcaagaaag gggctgaatg tttcgaggag ctctccaagt gcatgcaaga gaagtcaccg
     4921 cccttcagtg cggctgccct ggctggacac atggcacctg tgggacacct cccacctttt
     4981 agtcactctg gacacatcct acccacgccc acgcctatcc acccttcctc cagtctctct
     5041 tttggccacc cccacccgtc cagcatggtg actgccatgg gctagggatc cattcgatat
     5101 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     5161 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     5221 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     5281 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     5341 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     5401 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     5461 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     5521 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     5581 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     5641 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     5701 gcatcgatac cgtcgacctc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     5761 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
     5821 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     5881 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     5941 tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc agaactacac
     6001 accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     6061 tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt
     6121 gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg
     6181 cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt
     6241 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6301 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6361 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa
     6421 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     6481 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     6541 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     6601 acagcaaggg ggaggattgg gaagacaata gcaggcatgt gagcaaaagg ccagcaaaag
     6661 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6721 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6781 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6841 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6901 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6961 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7021 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7081 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     7141 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7201 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7261 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7321 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7381 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7441 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7501 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7561 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7621 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7681 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7741 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7801 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7861 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7921 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7981 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8041 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8101 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8161 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8221 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8281 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8341 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8401 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     8461 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     8521 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     8581 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     8641 ggcgttttgc gctgcttcg
//
