LOCUS       sequence-100301-        5984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a TALEN protein specific for mouse ROSA26 locus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5984)
  AUTHORS   Kasparek P, Krausova M, Haneckova R, Kriz V, Zbodakova O, Korinek V,
            Sedlacek R
  TITLE     Efficient gene targeting of the Rosa26 locus in mouse zygotes using 
            TALE nucleases.
  JOURNAL   FEBS Lett. 2014 Sep 18. pii: S0014-5793(14)00689-9. doi: 
            10.1016/j.febslet.2014.09.014.
  PUBMED    25241166
REFERENCE   2  (bases 1 to 5984)
  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: "FEBS Lett. 
            2014 Sep 18. pii: S0014-5793(14)00689-9. doi: 
            10.1016/j.febslet.2014.09.014."
COMMENT     Sequence Label: TALEN-mROSA26 KKR
FEATURES             Location/Qualifiers
     source          1..5984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        38..56
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      60..69
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             69..134
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             141..161
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3311..3432
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3322..3341)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3376..3395
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3565..3669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3670..3738,3739..4530)
                     /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     complement(3888..3907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4701..5289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5190..5209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5418..5775
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5418..5609
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5441..5461)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5626..5761
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5845..5977
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5960..5979
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 ctcccagttc aattacagct cttaaggcta gagtacttaa tacgactcac tataggctag
       61 ccaccatgga ctataaggac cacgacggag actacaagga tcatgatatt gattacaaag
      121 acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag
      181 cagccgtaga tttgagaact ttgggatatt cacagcagca gcaggaaaag atcaagccca
      241 aagtgaggtc gacagtcgcg cagcatcacg aagcgctggt gggtcatggg tttacacatg
      301 cccacatcgt agccttgtcg cagcaccctg cagcccttgg cacggtcgcc gtcaagtacc
      361 aggacatgat tgcggcgttg ccggaagcca cacatgaggc gatcgtcggt gtggggaaac
      421 agtggagcgg agcccgagcg cttgaggccc tgttgacggt cgcgggagag ctgagagggc
      481 ctccccttca gctggacacg ggccagttgc tgaagatcgc gaagcgggga ggagtcacgg
      541 cggtcgaggc ggtgcacgcg tggcgcaatg cactgacggg tgcccccctg aacctgaccc
      601 cggaccaagt ggtggctatc gccagccacg atggcggcaa gcaagcgctc gaaacggtgc
      661 agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa gtggtggcta
      721 tcgccagcaa caatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc
      781 tgtgccagga ccatggcctg accccggacc aagtggtggc tatcgccagc aacggtggcg
      841 gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc
      901 tgaccccgga ccaagtggtg gctatcgcca gcaacaatgg cggcaagcaa gcgctcgaaa
      961 cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg
     1021 tggctatcgc cagcaacatt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc
     1081 cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacg
     1141 gtggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc
     1201 atggcctgac tccggaccaa gtggtggcta tcgccagcca cgatggcggc aagcaagcgc
     1261 tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc
     1321 aagtggtggc tatcgccagc aacggtggcg gcaagcaagc gctcgaaacg gtgcagcggc
     1381 tgttgccggt gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca
     1441 gcaacaatgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc
     1501 aggaccatgg cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc
     1561 aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc
     1621 cggaccaagt ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc gaaacggtgc
     1681 agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa gtggtggcta
     1741 tcgccagcaa cattggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc
     1801 tgtgccagga ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg
     1861 gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc
     1921 tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa
     1981 cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgactccg gaccaagtgg
     2041 tggctatcgc cagccacgat ggcggcaagc aagcgctcga aacggtgcag cggctgttgc
     2101 cggtgctgtg ccaggaccat ggcctgactc cggaccaagt ggtggctatc gccagccacg
     2161 atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc
     2221 atggcctgac cccggaccaa gtggtggcta tcgccagcaa cattggcggc aagcaagcgc
     2281 tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc
     2341 aagtggtggc tatcgccagc aacaatggcg gcaagcaagc gctcgaaacg gtgcagcggc
     2401 tgttgccggt gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca
     2461 gcaacggtgg cggcaagcaa gcgctcgaaa gcattgtggc ccagctgagc cggcctgatc
     2521 cggcgttggc cgcgttgacc aacgaccacc tcgtagcgct ggcctgcctc ggcggacgac
     2581 ccgccttgga tgcggtgaag aaggggctcc cgcacgcgcc tgcattgatt aagcggacca
     2641 acagaaggat tcccgagagg acatcacatc gagtggcagg ttcccaactc gtgaagagtg
     2701 aacttgagga gaaaaagtcg gagctgcggc acaaattgaa atacgtaccg catgaataca
     2761 tcgaacttat cgaaattgct aggaactcga ctcaagacag aatccttgag atgaaggtaa
     2821 tggagttctt tatgaaggtt tatggatacc gagggaagca tctcggtgga tcacgaaaac
     2881 ccgacggagc aatctatacg gtggggagcc cgattgatta cggagtgatc gtcgacacga
     2941 aagcctacag cggtgggtac aatcttccca tcgggcaggc agatgagatg caacgttatg
     3001 tcaaagaaaa tcagaccagg aacaaacaca tcaatccaaa tgagtggtgg aaagtgtatc
     3061 cttcatcagt gaccgagttt aagtttttgt ttgtctctgg gcatttcaaa ggcaactata
     3121 aggcccagct cacacggttg aatcgcaaaa cgaactgcaa tggtgcggtt ttgtccgtag
     3181 aggaactgct cattggtgga gaaatgatca aagcgggaac tctgacactg gaagaagtca
     3241 gacgcaagtt taacaatggc gagatcaatt tccgctcata atctagagcg gccgcttcga
     3301 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3361 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3421 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     3481 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatcca
     3541 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3601 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3661 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3721 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3781 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3841 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3901 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3961 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4021 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4081 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4141 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4201 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4261 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4321 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4381 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4441 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4501 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4561 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4621 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4681 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4741 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4801 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4861 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4921 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4981 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5041 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5101 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5161 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5221 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5281 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5341 gctcacatgg ctcgacagat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     5401 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     5461 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5521 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5581 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5641 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5701 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5761 ctaggctttt gcaaaaagct tgattcttct gacacaacag tctcgaactt aagctgcaga
     5821 agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt ttaaggagac
     5881 caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt
     5941 ggtcttactg acatccactt tgcctttctc tccacaggtg tcca
//
