LOCUS       sequence-433433-        5962 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 5962)
  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 5962)
  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..5962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             34..60
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             435..446
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(2337..2361)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2384..2951
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2401..2953
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2545..2562)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2772..2791
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2965..2967,2968..2970,2971..3684)
                     /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(3010..3031)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3271..3290)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3618..3639
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3734..3833)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3945..3964)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3956..4765
                     /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(4023..4042)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4855..5443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5344..5363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5608..5694
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5786..5813
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     5821..5844
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    5863..5962
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggcgccggaa ccaattcagt cgacgccacc atgtatccgt atgatgtgcc ggattatgcg
       61 tttcctaacg cgccctacct gcccagctgc ctcgagagcc agcccgctat tcgcaatcag
      121 ggttacagca cggtcacctt cgacgggacg cccagctacg gtcacacgcc ctcgcaccat
      181 gcggcgcagt tccccaacca ctcattcaag catgaggatc ccatgggcca gcagggctcg
      241 ctgggtgagc agcagtactc ggtgccgccc ccggtctatg gctgccacac ccccaccgac
      301 agctgcaccg gcagccaggc tttgctgctg aggacgccct acagcagtga caatttatac
      361 caaatgacat cccagcttga atgcatgacc tggaatcaga tgaacttagg agccacctta
      421 aagggagttg ctgctgggag ctccagctca gtgaaatgga cagaagggca gagcaaccac
      481 agcacagggt acgagagcga taaccacaca acgcccatcc tctgcggagc ccaatacaga
      541 atacacacgc acggtgtctt cagaggcatt caggatgtgc gacgtgtgcc tggagtagcc
      601 ccgactcttg tacggtcggc atctgagacc agtgagaaac gccccttcat gtgtgcttac
      661 ccaggctgca ataagagata ttttaagctg tcccacttac agatgcacag caggaagcac
      721 actggtgaga aaccatacca gtgtgacttc aaggactgtg aacgaaggtt ttctcgttca
      781 gaccagctca aaagacacca aaggagacat acaggtgtga aaccattcca gtgtaaaact
      841 tgtcagcgaa agttctcccg gtccgaccac ctgaagaccc acaccaggac tcatacaggt
      901 aaaacaagtg aaaagccctt cagctgtcgg tggccaagtt gtcagaaaaa gtttgcccgg
      961 tcagatgaat tagtccgcca tcacaacatg catcagagaa acatgaccaa actccagctg
     1021 gcgctttgag gggtctccct cggggaccgt tcagtgtccc aggcagcaca gtgtgtgaac
     1081 tgctttcaag tctgactctc cactcctcct cactaaaaag gaagcttcag ttgatcttct
     1141 tcatccaact tccaagacaa gataccggtg cttctggaaa ctaccaggtg tgcctggaag
     1201 agttggtctc tgccctgcct acttttagtt gactcacagg ccctggagaa gcagctaaca
     1261 atgtctggtt agttaaaagc ccattgccat ttggtgtgga ttttctactg taagaagagc
     1321 catagctgat catgtccccc tgacccttcc cttctttttt tatgctcgtt ttcgctgggg
     1381 atggaattat tgtaccattt tctatcatgg aatatttata ggccagggca tgtgtatgtg
     1441 tctgctaatg taaactttgt catggtttcc atttactaac agcaacagca agaaataaat
     1501 cagagagcaa ggcatcgggg gtgaatcttg tctaacattc ccgaggtcag ccaggctgct
     1561 aacctggaaa gcaggatgta gttctgccag gcaactttta aagctcatgc atttcaagca
     1621 gctgaagaaa aaatcagaac taaccagtgc ctctgtatag aaatctaaaa gaattttacc
     1681 attcagttaa tccaatgtga acactggcac actgctctta agaaactatg aagatctgag
     1741 atttttttgt gtatgttttt gactcttttg agtggtaatc atatgtgtct ttatagatgt
     1801 acatacctcc ttgcacaaat ggaggggaat tcattttcat cactgggagt gtccttagtg
     1861 tataaaaacc atgctggtat atggcttcaa gttgtaaaaa tgaaagtggc tttaaaagaa
     1921 aataggggat ggtccaggat ctccactgat aagactgttt ttaagtaact taaggacctt
     1981 tgggtctaca agtatatgtg aaaaaaatga gacttactgg gtgaggaaat ccattgttta
     2041 aagatggtcg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg ttgtgttgcg ttttgttttt
     2101 taagggaggg aatttattat ttaccgttgc ttgaaattac tgtgtaaata tatgtctgat
     2161 aatgatttgc tctttgacaa ctaaaattag gactgtataa gtactagatg catcactggg
     2221 tgttggtctt acaagatatt gatgataaca cttaaaattg taacctgcat ttttcacttt
     2281 gctctcaatt aaagtctatt caaaaggtaa ttcgcggccg cactcgagat atctagaccc
     2341 agctttcttg tacaaagtgg tgattccccc gggctgcagg aattccgccc ccccccccta
     2401 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     2461 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     2521 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     2581 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     2641 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     2701 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     2761 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     2821 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     2881 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     2941 cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt caccggggtg
     3001 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtctggc
     3061 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3121 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3181 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3241 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3301 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3361 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3421 atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc
     3481 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3541 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3601 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3661 ggcatggacg agctgtacaa gtaatgaatt aattaagaat tatcaagctt atcgataccg
     3721 tcgagatatc tagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     3781 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     3841 gctgcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     3901 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     3961 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     4021 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     4081 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     4141 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     4201 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     4261 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     4321 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     4381 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     4441 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     4501 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     4561 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     4621 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     4681 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     4741 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac attattcaga
     4801 ttgggccccg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4861 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4921 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4981 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5041 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5101 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5161 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5221 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5281 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5341 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5401 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5461 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5521 ccctgattct gtggataacc gtattaccgc tagcatggat ctcggggacg tctaactact
     5581 aagcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc ggaagactgg
     5641 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     5701 ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     5761 taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct ttttgcgttt
     5821 ctacaaactc ttcctgttag ttagttactt aagctcgggc cccaaataat gattttattt
     5881 tgactgatag tgacctgttc gttgcaacaa attgataagc aatgcttttt tataatgcca
     5941 actttgtaca aaaaagcagg ct
//
