LOCUS       sequence-433654-        5947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector with WT1 -KTS mutant with N 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 N terminal HA
FEATURES             Location/Qualifiers
     source          1..5947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      120..206
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      298..325
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     333..356
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    375..474
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             508..534
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             909..920
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(2796..2820)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2843..3410
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2860..3412
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3004..3021)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3231..3250
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3424..3426,3427..3429,3430..4143)
                     /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(3469..3490)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3730..3749)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4077..4098
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4193..4292)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4404..4423)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4415..5224
                     /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(4482..4501)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5314..5902
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5803..5822
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
       61 gctagcatgg atctcgggga cgtctaacta ctaagcgaga gtagggaact gccaggcatc
      121 aaataaaacg aaaggctcag tcggaagact gggcctttcg ttttatctgt tgtttgtcgg
      181 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gtgaagcaac
      241 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa actaagcaga
      301 aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttcctgtt agttagttac
      361 ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt tcgttgcaac
      421 aaattgataa gcaatgcttt tttataatgc caactttgta caaaaaagca ggctggcgcc
      481 ggaaccaatt cagtcgacgc caccatgtat ccgtatgatg tgccggatta tgcgtttcct
      541 aacgcgccct acctgcccag ctgcctcgag agccagcccg ctattcgcaa tcagggttac
      601 agcacggtca ccttcgacgg gacgcccagc tacggtcaca cgccctcgca ccatgcggcg
      661 cagttcccca accactcatt caagcatgag gatcccatgg gccagcaggg ctcgctgggt
      721 gagcagcagt actcggtgcc gcccccggtc tatggctgcc acacccccac cgacagctgc
      781 accggcagcc aggctttgct gctgaggacg ccctacagca gtgacaattt ataccaaatg
      841 acatcccagc ttgaatgcat gacctggaat cagatgaact taggagccac cttaaaggga
      901 gttgctgctg ggagctccag ctcagtgaaa tggacagaag ggcagagcaa ccacagcaca
      961 gggtacgaga gcgataacca cacaacgccc atcctctgcg gagcccaata cagaatacac
     1021 acgcacggtg tcttcagagg cattcaggat gtgcgacgtg tgcctggagt agccccgact
     1081 cttgtacggt cggcatctga gaccagtgag aaacgcccct tcatgtgtgc ttacccaggc
     1141 tgcaataaga gatattttaa gctgtcccac ttacagatgc acagcaggaa gcacactggt
     1201 gagaaaccat accagtgtga cttcaaggac tgtgaacgaa ggttttctcg ttcagaccag
     1261 ctcaaaagac accaaaggag acatacaggt gtgaaaccat tccagtgtaa aacttgtcag
     1321 cgaaagttct cccggtccga ccacctgaag acccacacca ggactcatac aggtgaaaag
     1381 cccttcagct gtcggtggcc aagttgtcag aaaaagtttg cccggtcaga tgaattagtc
     1441 cgccatcaca acatgcatca gagaaacatg accaaactcc agctggcgct ttgaggggtc
     1501 tccctcgggg accgttcagt gtcccaggca gcacagtgtg tgaactgctt tcaagtctga
     1561 ctctccactc ctcctcacta aaaaggaaac ttcagttgat cttcttcatc caacttccaa
     1621 gacaagatac cggtgcttct ggaaactacc aggtgtgcct ggaagagttg gtctctgccc
     1681 tgcctacttt tagttgactc acaggccctg gagaagcagc taacaatgtc tggttagtta
     1741 aaagcccatt gccatttggt gtggattttc tactgtaaga agagccatag ctgatcatgt
     1801 ccccctgacc cttcccttct ttttttatgc tcgttttcgc tggggatgga attattgtac
     1861 cattttctat catggaatat ttataggcca gggcatgtgt atgtgtctgc taatgtaaac
     1921 tttgtcatgg tttccattta ctaacagcaa cagcaagaaa taaatcagag agcaaggcat
     1981 cgggggtgaa tcttgtctaa cattcccgag gtcagccagg ctgctaacct ggaaagcagg
     2041 atgtagttct gccaggcaac ttttaaagct catgcatttc aagcagctga agaaagaatc
     2101 agaactaacc agtacctctg tatagaaatc taaaagaatt ttaccattca gttaattcaa
     2161 tgtgaacact ggcacactgc tcttaagaaa ctatgaagat ctgagatttt tttgtgtatg
     2221 tttttgactc ttttgagtgg taatcatatg tgtctttata gatgtacata cctccttgca
     2281 caaacggagg ggaattcatt ttcatcactg ggagtgtcct tagtgtataa aaaccatgct
     2341 ggtatatggc ttcaagttgt aaaaatgaaa gcgactttaa aagaaaatag gggatggtcc
     2401 aggatctcca ctgataagac tgtttttaag taacttaagg acctttgggt ctacaagtat
     2461 atgtgaaaaa aatgagactt actgggtgag gaaatccatt gtttaaagat ggtcgtgtgt
     2521 gtgtgtgtgt gtgtgtgtgt tgtgttgtgt tttgtttttt aagggaggga gtttattatt
     2581 taccgttgct tgaaattact gtgtaaatat atgtctgata atgatttgct ctttgacaac
     2641 taaaattagg actgtataag tactagatgc atcactgggt gttgatctta caagatattg
     2701 atgataacac ttaaaattgt aacctgcatt tttcactttg ctctcaatta aagtctattc
     2761 aaaaggtaat tcgcggccgc actcgagata tctagaccca gctttcttgt acaaagtggt
     2821 gattcccccg ggctgcagga attccgcccc ccccccctaa cgttactggc cgaagccgct
     2881 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     2941 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     3001 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     3061 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     3121 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     3181 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     3241 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     3301 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     3361 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     3421 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3481 gacggcgacg taaacggcca caagttcagc gtgtctggcg agggcgaggg cgatgccacc
     3541 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     3601 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     3661 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3721 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3781 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3841 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     3901 aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     3961 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     4021 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     4081 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     4141 taatgaatta attaagaatt atcaagctta tcgataccgt cgagatatct agacccagct
     4201 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     4261 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagctct ggcccgtgtc
     4321 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     4381 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     4441 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     4501 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     4561 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     4621 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     4681 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     4741 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     4801 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     4861 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     4921 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     4981 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     5041 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     5101 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     5161 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     5221 ctaatcagaa ttggttaatt ggttgtaaca ttattcagat tgggccccgt tccactgagc
     5281 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5341 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5401 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5461 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5521 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5581 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5641 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5701 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5761 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5821 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5881 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5941 ttgctgg
//
