LOCUS       sequence-60741-e        7579 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7579)
  TITLE     Plasmids to assay for nuclease cleavage in planta
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7579)
  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            "Unpublished"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: pHEX2 T494 TALENs
FEATURES             Location/Qualifiers
     source          1..7579
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             39..104
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             111..131
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3975..4028
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4035..4100
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4107..4127
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6960..7547
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMER
                     YVEENQTRNKHLNPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     CDS             7550..7561
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 gtacaaaaaa gcaggctccg aattcgccct tcaccatgga ttataaggat cacgatggcg
       61 actacaagga ccacgatatt gactacaaag acgatgacga taaaatggct cctaagaaaa
      121 agcgcaaagt cggtatccat ggcgttccct ctagaatggt ggatctacgc acgctcggct
      181 acagtcagca gcagcaagag aagatcaaac cgaaggtgcg ttcgacagtg gcgcagcacc
      241 acgaggcact ggtgggccat gggtttacac acgcgcacat cgttgcgctc agccaacacc
      301 cggcagcgtt agggaccgtc gctgtcacgt atcagcacat aatcacggcg ttgccagagg
      361 cgacacacga agacatcgtt ggcgtcggca aacagtggtc cggcgcacgc gccctggagg
      421 ccttgctcac ggatgcgggg gagttgagag gtccgccgtt acagttggac acaggccaac
      481 ttgtgaagat tgcaaaacgt ggcggcgtga ccgcaatgga ggcagtgcat gcatcgcgca
      541 atgcactgac gggtgccccc ctgaacctga ccccggacca agtggtggct atcgccagca
      601 acaatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg
      661 accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc ggcaagcaag
      721 cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg
      781 accaagtggt ggctatcgcc agcaacattg gcggcaagca agcgctcgaa acggtgcagc
      841 ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg gtggctatcg
      901 ccagcaacat tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt
      961 gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac aatggcggca
     1021 agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga
     1081 ccccggacca agtggtggct atcgccagca acaatggcgg caagcaagcg ctcgaaacgg
     1141 tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg
     1201 ctatcgccag caacattggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg
     1261 tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc agccacgatg
     1321 gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg
     1381 gcctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag caagcgctcg
     1441 aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc ccggaccaag
     1501 tggtggctat cgccagcaac attggcggca agcaagcgct cgaaacggtg cagcggctgt
     1561 tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct atcgccagcc
     1621 acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg
     1681 accatggcct gaccccggac caagtggtgg ctatcgccag caacattggc ggcaagcaag
     1741 cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg
     1801 accaagtggt ggctatcgcc agcaacattg gcggcaagca agcgctcgaa acggtgcagc
     1861 ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg gtggctatcg
     1921 ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt
     1981 gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac attggcggca
     2041 agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga
     2101 ccccggacca agtggtggct atcgccagca acattggcgg caagcaagcg ctcgaaacgg
     2161 tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg
     2221 ctatcgccag caacaatggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg
     2281 tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc agcaacaatg
     2341 gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg
     2401 gcctgactcc ggaccaagtg gtggctatcg ccagccacga tggcggcaag caagcgctcg
     2461 aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact ccggaccaag
     2521 tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg cagcggctgt
     2581 tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct atcgccagca
     2641 acattggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg
     2701 accatggcct gaccccggac caagtggtgg ctatcgccag caacattggc ggcaagcaag
     2761 cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg
     2821 accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc
     2881 ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg gtggctatcg
     2941 ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt
     3001 gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac attggcggca
     3061 agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga
     3121 ccccggacca agtggtggct atcgccagca acggtggcgg caagcaagcg ctcgaaagca
     3181 ttgtggccca gctgagccgg cctgatccgg cgttggccgc gttgaccaac gaccacctcg
     3241 tcgccttggc ctgcctcggc ggacgtcctg ccatggatgc agtgaaaaag ggattgccgc
     3301 acgcgccgga attgatcaga agagtcaatc gccgtattgg cgaacgcacg tcccatcgcg
     3361 ttgccggatc ccagctggtg aagtccgagc tggaagaaaa aaagagcgag ctgcgccaca
     3421 agctcaagta cgtgccccac gagtacatcg agctgatcga gatcgcccgc aacagcaccc
     3481 aagaccgcat cctggagatg aaagtgatgg agttcttcat gaaggtgtac ggctaccgcg
     3541 gcaagcacct gggcggctcc cgcaagcccg atggcgccat ctacaccgtg ggctccccca
     3601 tcgactatgg cgtcattgtc gacaccaagg cctactccgg cggctacaac ttacccatcg
     3661 gtcaggccga cgagatgcaa cgctacgtga aggagaacca gacccgcaat aagcacatta
     3721 atcccaacga gtggtggaag gtgtacccct cctccgtgac cgagttcaaa ttcctgttcg
     3781 tgtccggcca cttcaagggc aattataagg cccaactgac ccgcctgaac cacaagacca
     3841 actgcaacgg cgccgtgctg tccgtggagg aactgctgat cggcggcgag atgatcaagg
     3901 ctggtaccct gaccctggaa gaggtgcgcc gcaagttcaa caatggtgaa atcaatttca
     3961 ggtccggcgg cggagagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc
     4021 ccggccctag gatggactac aaagaccatg acggtgatta taaagatcat gacatcgatt
     4081 acaaggatga cgatgacaag atggccccca agaagaagag gaaggtgggc attcacgggg
     4141 tgccggctag aatggtggat ctacgcacgc tcggctacag tcagcagcag caagagaaga
     4201 tcaaaccgaa ggtgcgttcg acagtggcgc agcaccacga ggcactggtg ggccatgggt
     4261 ttacacacgc gcacatcgtt gcgctcagcc aacacccggc agcgttaggg accgtcgctg
     4321 tcacgtatca gcacataatc acggcgttgc cagaggcgac acacgaagac atcgttggcg
     4381 tcggcaaaca gtggtccggc gcacgcgccc tggaggcctt gctcacggat gcgggggagt
     4441 tgagaggtcc gccgttacag ttggacacag gccaacttgt gaagattgca aaacgtggcg
     4501 gcgtgaccgc aatggaggca gtgcatgcat cgcgcaatgc actgacgggt gcccccctga
     4561 acctgacccc ggaccaagtg gtggctatcg ccagccacga tggcggcaag caagcgctcg
     4621 aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact ccggaccaag
     4681 tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg cagcggctgt
     4741 tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct atcgccagca
     4801 acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg
     4861 accatggcct gaccccggac caagtggtgg ctatcgccag caacggtggc ggcaagcaag
     4921 cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg
     4981 accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc
     5041 ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg gtggctatcg
     5101 ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt
     5161 gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac gatggcggca
     5221 agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga
     5281 ccccggacca agtggtggct atcgccagca acattggcgg caagcaagcg ctcgaaacgg
     5341 tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg
     5401 ctatcgccag caacggtggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg
     5461 tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc agcaacaatg
     5521 gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg
     5581 gcctgacccc ggaccaagtg gtggctatcg ccagcaacaa tggcggcaag caagcgctcg
     5641 aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc ccggaccaag
     5701 tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg cagcggctgt
     5761 tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct atcgccagca
     5821 acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg
     5881 accatggcct gaccccggac caagtggtgg ctatcgccag caacaatggc ggcaagcaag
     5941 cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg
     6001 accaagtggt ggctatcgcc agcaacaatg gcggcaagca agcgctcgaa acggtgcagc
     6061 ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg gtggctatcg
     6121 ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt
     6181 gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac gatggcggca
     6241 agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga
     6301 ccccggacca agtggtggct atcgccagca acggtggcgg caagcaagcg ctcgaaacgg
     6361 tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg
     6421 ctatcgccag caacggtggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg
     6481 tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc agcaacaatg
     6541 gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg
     6601 gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag caagcgctcg
     6661 aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc ccggaccaag
     6721 tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaagcatt gtggcccagc
     6781 tgagccggcc tgatccggcg ttggccgcgt tgaccaacga ccacctcgtc gccttggcct
     6841 gcctcggcgg acgtcctgcc atggatgcag tgaaaaaggg attgccgcac gcgccggaat
     6901 tgatcagaag agtcaatcgc cgtattggcg aacgcacgtc ccatcgcgtt gccggatctc
     6961 aactagtcaa aagtgaactg gaggagaaga aatctgaact tcgtcataaa ttgaaatatg
     7021 tgcctcatga atatattgaa ttaattgaaa ttgccagaaa ttccactcag gatagaattc
     7081 ttgaaatgaa ggtaatggaa ttttttatga aagtttatgg atatagaggt aaacatttgg
     7141 gtggatcaag gaaaccggac ggagcaattt atactgtcgg atctcctatt gattacggtg
     7201 tgatcgtgga tactaaagct tatagcggag gttataatct gccaattggc caagcagatg
     7261 aaatggagcg atatgtcgaa gaaaatcaaa cacgaaacaa acatctcaac cctaatgaat
     7321 ggtggaaagt ctatccatct tctgtaacgg aatttaagtt tttatttgtg agtggtcact
     7381 ttaaaggaaa ctacaaagct cagcttacac gattaaatca tatcactaat tgtaatggag
     7441 ctgttcttag tgtagaagag cttttaattg gtggagaaat gattaaagcc ggcacattaa
     7501 ccttagagga agtgagacgg aaatttaata acggcgagat aaacttttaa tagaagggcg
     7561 aattcgaccc agctttctt
//
