LOCUS       sequence-358856-        9562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLT3-ITD (internal tandem duplication) constitutively 
            active mutant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9562)
  TITLE     Mammalian expression plasmids for FLT3 wildtype and common AML 
            mutant FLT3 proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9562)
  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: 
            "Unpublished"
COMMENT     Sequence Label: FLT3-ITD-MSCV-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..9562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     173..191
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(229..251)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     351..370
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     564..586
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     714..733
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             933..1449
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1513..1854
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1921..2337
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2267..2289
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2305..2321
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    2363..2387
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5451..5475)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    5487..6054
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5504..6056
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5648..5665)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5875..5894
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6068..6070,6071..6073,6074..6787)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6113..6134)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6374..6393)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6721..6742
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             6889..7403
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7565..7581
                     /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)."
     promoter        complement(join(7589..7595,7596..7613,7614..7619))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7634..7655
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7772..7789)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7943..8531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8023..8042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8702..9493,9494..9562))
                     /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     9325..9344
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
      181 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
      241 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
      301 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
      361 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
      421 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
      481 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
      541 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat
      601 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
      661 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
      721 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
      781 ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta
      841 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
      901 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
      961 aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga
     1021 agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt
     1081 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg
     1141 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     1201 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     1261 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
     1321 agactgcgtc gcccgggtac ccgtattccc aataaagcct cttgctgttt gcatccgaat
     1381 cgtggactcg ctgatccttg ggagggtctc ctcagattga ttgactgccc acctcggggg
     1441 tctttcattt ggaggttcca ccgagatttg gagacccctg cccagggacc accgaccccc
     1501 ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt
     1561 tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt tagctaacta gctctgtatc
     1621 tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt
     1681 cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat
     1741 ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct
     1801 ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc
     1861 tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag
     1921 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
     1981 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
     2041 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
     2101 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
     2161 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
     2221 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
     2281 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga
     2341 attagatctc tcgaggttat caacaagttt gtacaaaaaa gcaggcttta aaggaaccaa
     2401 ttcagtcgac tggatccatg ccggcgttgg cgcgcgacgg cggccagctg ccgctgctcg
     2461 ttgttttttc tgcaatgata tttgggacta ttacaaatca agatctgcct gtgatcaagt
     2521 gtgttttaat caatcataag aacaatgatt catcagtggg gaagtcatca tcatatccca
     2581 tggtatcaga atccccggaa gacctcgggt gtgcgttgag accccagagc tcagggacag
     2641 tgtacgaagc tgccgctgtg gaagtggatg tatctgcttc catcacactg caagtgctgg
     2701 tcgatgcccc agggaacatt tcctgtctct gggtctttaa gcacagctcc ctgaattgcc
     2761 agccacattt tgatttacaa aacagaggag ttgtttccat ggtcattttg aaaatgacag
     2821 aaacccaagc tggagaatac ctacttttta ttcagagtga agctaccaat tacacaatat
     2881 tgtttacagt gagtataaga aataccctgc tttacacatt aagaagacct tactttagaa
     2941 aaatggaaaa ccaggacgcc ctggtctgca tatctgagag cgttccagag ccgatcgtgg
     3001 aatgggtgct ttgcgattca cagggggaaa gctgtaaaga agaaagtcca gctgttgtta
     3061 aaaaggagga aaaagtgctt catgaattat ttgggatgga cataaggtgc tgtgccagaa
     3121 atgaactggg cagggaatgc accaggctgt tcacaataga tctaaatcaa actcctcaga
     3181 ccacattgcc acaattattt cttaaagtag gggaaccctt atggataagg tgcaaagctg
     3241 ttcatgtgaa ccatggattc gggctcacct gggaattaga aaacaaagca ctcgaggagg
     3301 gcaactactt tgagatgagt acctattcaa caaacagaac tatgatacgg attctgtttg
     3361 cttttgtatc atcagtggca agaaacgaca ccggatacta cacttgttcc tcttcaaagc
     3421 atcccagtca atcagctttg gttaccatcg tagaaaaggg atttataaat gctaccaatt
     3481 caagtgaaga ttatgaaatt gaccaatatg aagagttttg tttttctgtc aggtttaaag
     3541 cctacccaca aatcagatgt acgtggacct tctctcgaaa atcatttcct tgtgagcaaa
     3601 agggtcttga taacggatac agcatatcca agttttgcaa tcataagcac cagccaggag
     3661 aatatatatt ccatgcagaa aatgatgatg cccaatttac caaaatgttc acgctgaata
     3721 taagaaggaa acctcaagtg ctcgcagaag catcggcaag tcaggcgtcc tgtttctcgg
     3781 atggataccc attaccatct tggacctgga agaagtgttc agacaagtct cccaactgca
     3841 cagaagagat cacagaagga gtctggaata gaaaggctaa cagaaaagtg tttggacagt
     3901 gggtgtcgag cagtactcta aacatgagtg aagccataaa agggttcctg gtcaagtgct
     3961 gtgcatacaa ttcccttggc acatcttgtg agacgatcct tttaaactct ccaggcccct
     4021 tccctttcat ccaagacaac atctcattct atgcaacaat tggtgtttgt ctcctcttca
     4081 ttgtcgtttt aaccctgcta atttgtcaca agtacaaaaa gcaatttagg tatgaaagcc
     4141 agctacagat ggtacaggtg accggctcct cagataatga gtacttctac gttgatttca
     4201 gagaatatga atatgatctc agagaatatg aatatgatct caaatgggag tttccaagag
     4261 aaaatttaga gtttgggaag gtactaggat caggtgcttt tggaaaagtg atgaacgcaa
     4321 cagcttatgg aattagcaaa acaggagtct caatccaggt tgccgtcaaa atgctgaaag
     4381 aaaaagcaga cagctctgaa agagaggcac tcatgtcaga actcaagatg atgacccagc
     4441 tgggaagcca cgagaatatt gtgaacctgc tgggggcgtg cacactgtca ggaccaattt
     4501 acttgatttt tgaatactgt tgctatggtg atcttctcaa ctatctaaga agtaaaagag
     4561 aaaaatttca caggacttgg acagagattt tcaaggaaca caatttcagt ttttacccca
     4621 ctttccaatc acatccaaat tccagcatgc ctggttcaag agaagttcag atacacccgg
     4681 actcggatca aatctcaggg cttcatggga attcatttca ctctgaagat gaaattgaat
     4741 atgaaaacca aaaaaggctg gaagaagagg aggacttgaa tgtgcttaca tttgaagatc
     4801 ttctttgctt tgcatatcaa gttgccaaag gaatggaatt tctggaattt aagtcgtgtg
     4861 ttcacagaga cctggccgcc aggaacgtgc ttgtcaccca cgggaaagtg gtgaagatat
     4921 gtgactttgg attggctcga gatatcatga gtgattccaa ctatgttgtc aggggcaatg
     4981 cccgtctgcc tgtaaaatgg atggcccccg aaagcctgtt tgaaggcatc tacaccatta
     5041 agagtgatgt ctggtcatat ggaatattac tgtgggaaat cttctcactt ggtgtgaatc
     5101 cttaccctgg cattccggtt gatgctaact tctacaaact gattcaaaat ggatttaaaa
     5161 tggatcagcc attttatgct acagaagaaa tatacattat aatgcaatcc tgctgggctt
     5221 ttgactcaag gaaacggcca tccttcccta atttgacttc gtttttagga tgtcagctgg
     5281 cagatgcaga agaagcgatg tatcagaatg tggatggccg tgtttcggaa tgtcctcaca
     5341 cctaccaaaa caggcgacct ttcagcagag agatggattt ggggctactc tctccgcagg
     5401 ctcaggtcga agattcgtag atctagagcg gccgcactcg agatatctag acccagcttt
     5461 cttgtacaaa gtggttgata acgaattccg cccccccccc ctaacgttac tggccgaagc
     5521 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     5581 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     5641 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     5701 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     5761 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     5821 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     5881 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     5941 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     6001 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     6061 cacaaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     6121 gctggacggc gacgtaaacg gccacaagtt cagcgtgtct ggcgagggcg agggcgatgc
     6181 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     6241 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     6301 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     6361 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     6421 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     6481 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     6541 gaagaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     6601 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     6661 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     6721 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     6781 caagtaatga attaattaag aattatcaag cttatcgata ccgtcgacct gcagccaagc
     6841 ttatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     6901 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat
     6961 aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc
     7021 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     7081 ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     7141 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     7201 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg
     7261 cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa accctcttgc
     7321 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     7381 acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa caacattctg
     7441 agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt gaatccacat
     7501 actccaatac tcctgaaata gttcattatg gacagcgcag aaagagctgg ggagaattgt
     7561 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7621 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7681 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     7741 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7801 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7861 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     7921 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     7981 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8041 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     8101 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     8161 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     8221 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     8281 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     8341 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     8401 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     8461 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     8521 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8581 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     8641 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     8701 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8761 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8821 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     8881 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     8941 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9001 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     9061 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     9121 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     9181 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     9241 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     9301 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     9361 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     9421 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     9481 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9541 cacggaaatg ttgaatactc at
//
