LOCUS       sequence-433565-        5947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector with WT1 -KTS mutant with C terminal HA for gateway 
            cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5947)
  AUTHORS   Potluri S, Assi SA, Chin PS, Coleman DJL, Pickin A, Moriya S, Seki 
            N, Heidenreich O, Cockerill PN, Bonifer C
  TITLE     Isoform-specific and signaling-dependent propagation of acute 
            myeloid leukemia by Wilms tumor 1.
  JOURNAL   Cell Rep. 2021 Apr 20;35(3):109010. doi: 
            10.1016/j.celrep.2021.109010.
  PUBMED    33882316
REFERENCE   2  (bases 1 to 5947)
  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: "Cell Rep.";
            date: "2021-04-20"; volume: "35(3):109010. doi"; pages: " 
            10.1016/j.celrep.2021.109010"
COMMENT     Sequence Label: pENTR-GFP-WT1 -KTS C terminal HA
FEATURES             Location/Qualifiers
     source          1..5947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     282..301
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      546..632
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      724..751
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     759..782
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    801..900
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1308..1319
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1832..1858
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(3222..3246)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3269..3836
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3286..3838
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3430..3447)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3657..3676
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3850..3852,3853..3855,3856..4569)
                     /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(3895..3916)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4156..4175)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4503..4524
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4619..4718)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4830..4849)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4841..5650
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4908..4927)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(5740..5947,1..381)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
       61 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      121 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
      181 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
      241 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
      301 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
      361 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
      421 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
      481 attaccgcta gcatggatct cggggacgtc taactactaa gcgagagtag ggaactgcca
      541 ggcatcaaat aaaacgaaag gctcagtcgg aagactgggc ctttcgtttt atctgttgtt
      601 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgtga
      661 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaacta
      721 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt cctgttagtt
      781 agttacttaa gctcgggccc caaataatga ttttattttg actgatagtg acctgttcgt
      841 tgcaacaaat tgataagcaa tgctttttta taatgccaac tttgtacaaa aaagcaggct
      901 ggcgccggaa ccaattcagt cgacgccacc atgtttccta acgcgcccta cctgcccagc
      961 tgcctcgaga gccagcccgc tattcgcaat cagggttaca gcacggtcac cttcgacggg
     1021 acgcccagct acggtcacac gccctcgcac catgcggcgc agttccccaa ccactcattc
     1081 aagcatgagg atcccatggg ccagcagggc tcgctgggtg agcagcagta ctcggtgccg
     1141 cccccggtct atggctgcca cacccccacc gacagctgca ccggcagcca ggctttgctg
     1201 ctgaggacgc cctacagcag tgacaattta taccaaatga catcccagct tgaatgcatg
     1261 acctggaatc agatgaactt aggagccacc ttaaagggag ttgctgctgg gagctccagc
     1321 tcagtgaaat ggacagaagg gcagagcaac cacagcacag ggtacgagag cgataaccac
     1381 acaacgccca tcctctgcgg agcccaatac agaatacaca cgcacggtgt cttcagaggc
     1441 attcaggatg tgcgacgtgt gcctggagta gccccgactc ttgtacggtc ggcatctgag
     1501 accagtgaga aacgcccctt catgtgtgct tacccaggct gcaataagag atattttaag
     1561 ctgtcccact tacagatgca cagcaggaag cacactggtg agaaaccata ccagtgtgac
     1621 ttcaaggact gtgaacgaag gttttctcgt tcagaccagc tcaaaagaca ccaaaggaga
     1681 catacaggtg tgaaaccatt ccagtgtaaa acttgtcagc gaaagttctc ccggtccgac
     1741 cacctgaaga cccacaccag gactcataca ggtgaaaagc ccttcagctg tcggtggcca
     1801 agttgtcaga aaaagtttgc ccggtcagat gtatccgtat gatgtgccgg attatgcgaa
     1861 ttagtccgcc atcacaacat gcatcagaga aacatgacca aactccagct ggcgctttga
     1921 ggggtctccc tcggggaccg ttcagtgtcc caggcagcac agtgtgtgaa ctgctttcaa
     1981 gtctgactct ccactcctcc tcactaaaaa ggaaacttca gttgatcttc ttcatccaac
     2041 ttccaagaca agataccggt gcttctggaa actaccaggt gtgcctggaa gagttggtct
     2101 ctgccctgcc tacttttagt tgactcacag gccctggaga agcagctaac aatgtctggt
     2161 tagttaaaag cccattgcca tttggtgtgg attttctact gtaagaagag ccatagctga
     2221 tcatgtcccc ctgacccttc ccttcttttt ttatgctcgt tttcgctggg gatggaatta
     2281 ttgtaccatt ttctatcatg gaatatttat aggccagggc atgtgtatgt gtctgctaat
     2341 gtaaactttg tcatggtttc catttactaa cagcaacagc aagaaataaa tcagagagca
     2401 aggcatcggg ggtgaatctt gtctaacatt cccgaggtca gccaggctgc taacctggaa
     2461 agcaggatgt agttctgcca ggcaactttt aaagctcatg catttcaagc agctgaagaa
     2521 agaatcagaa ctaaccagta cctctgtata gaaatctaaa agaattttac cattcagtta
     2581 attcaatgtg aacactggca cactgctctt aagaaactat gaagatctga gatttttttg
     2641 tgtatgtttt tgactctttt gagtggtaat catatgtgtc tttatagatg tacatacctc
     2701 cttgcacaaa cggaggggaa ttcattttca tcactgggag tgtccttagt gtataaaaac
     2761 catgctggta tatggcttca agttgtaaaa atgaaagcga ctttaaaaga aaatagggga
     2821 tggtccagga tctccactga taagactgtt tttaagtaac ttaaggacct ttgggtctac
     2881 aagtatatgt gaaaaaaatg agacttactg ggtgaggaaa tccattgttt aaagatggtc
     2941 gtgtgtgtgt gtgtgtgtgt gtgtgttgtg ttgtgttttg ttttttaagg gagggagttt
     3001 attatttacc gttgcttgaa attactgtgt aaatatatgt ctgataatga tttgctcttt
     3061 gacaactaaa attaggactg tataagtact agatgcatca ctgggtgttg atcttacaag
     3121 atattgatga taacacttaa aattgtaacc tgcatttttc actttgctct caattaaagt
     3181 ctattcaaaa ggtaattcgc ggccgcactc gagatatcta gacccagctt tcttgtacaa
     3241 agtggtgatt cccccgggct gcaggaattc cgcccccccc ccctaacgtt actggccgaa
     3301 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     3361 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     3421 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     3481 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     3541 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     3601 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     3661 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     3721 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     3781 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg
     3841 gccacaacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3901 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg cgagggcgat
     3961 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     4021 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     4081 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4141 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     4201 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4261 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     4321 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcagcgtg
     4381 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     4441 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     4501 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     4561 tacaagtaat gaattaatta agaattatca agcttatcga taccgtcgag atatctagac
     4621 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     4681 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctgc agctctggcc
     4741 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     4801 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     4861 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     4921 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     4981 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     5041 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     5101 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     5161 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     5221 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     5281 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     5341 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     5401 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     5461 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     5521 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     5581 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     5641 gtttttctaa tcagaattgg ttaattggtt gtaacattat tcagattggg ccccgttcca
     5701 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5761 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5821 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5881 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5941 tacatac
//
