LOCUS       sequence-100306-        5780 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 5780)
  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 5780)
  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 ELD
FEATURES             Location/Qualifiers
     source          1..5780
                     /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    3107..3228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3118..3137)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3172..3191
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3361..3465
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3466..3534,3535..4326)
                     /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(3684..3703)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4497..5085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4986..5005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5214..5571
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5214..5405
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5237..5257)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5422..5557
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5641..5773
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5756..5775
                     /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 gccagcaaca atggcggcaa gcaagcgctc gaaacggtgc
      661 agcggctgtt gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta
      721 tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc
      781 tgtgccagga ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg
      841 gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc
      901 tgactccgga ccaagtggtg gctatcgcca gccacgatgg cggcaagcaa gcgctcgaaa
      961 cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg
     1021 tggctatcgc cagcaacatt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc
     1081 cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaaca
     1141 atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc
     1201 atggcctgac cccggaccaa gtggtggcta tcgccagcaa cattggcggc aagcaagcgc
     1261 tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc
     1321 aagtggtggc tatcgccagc aacattggcg gcaagcaagc gctcgaaacg gtgcagcggc
     1381 tgttgccggt gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca
     1441 gcaacaatgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc
     1501 aggaccatgg cctgaccccg gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc
     1561 aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc
     1621 cggaccaagt ggtggctatc gccagccacg atggcggcaa gcaagcgctc gaaacggtgc
     1681 agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa gtggtggcta
     1741 tcgccagcaa cggtggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc
     1801 tgtgccagga ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg
     1861 gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc
     1921 tgactccgga ccaagtggtg gctatcgcca gccacgatgg cggcaagcaa gcgctcgaaa
     1981 cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgactccg gaccaagtgg
     2041 tggctatcgc cagccacgat ggcggcaagc aagcgctcga aacggtgcag cggctgttgc
     2101 cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaaca
     2161 atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc
     2221 atggcctgac cccggaccaa gtggtggcta tcgccagcca cgatggcggc aagcaagcgc
     2281 tcgaaagcat tgtggcccag ctgagccggc ctgatccggc gttggccgcg ttgaccaacg
     2341 accacctcgt agcgctggcc tgcctcggcg gacgacccgc cttggatgcg gtgaagaagg
     2401 ggctcccgca cgcgcctgca ttgattaagc ggaccaacag aaggattccc gagaggacat
     2461 cacatcgagt ggcaggttcc caactcgtga agagtgaact tgaggagaaa aagtcggagc
     2521 tgcggcacaa attgaaatac gtaccgcatg aatacatcga acttatcgaa attgctagga
     2581 actcgactca agacagaatc cttgagatga aggtaatgga gttctttatg aaggtttatg
     2641 gataccgagg gaagcatctc ggtggatcac gaaaacccga cggagcaatc tatacggtgg
     2701 ggagcccgat tgattacgga gtgatcgtcg acacgaaagc ctacagcggt gggtacaatc
     2761 ttcccatcgg gcaggcagat gagatggaac gttatgtcga agaaaatcag accagggaca
     2821 aacacctcaa tccaaatgag tggtggaaag tgtatccttc atcagtgacc gagtttaagt
     2881 ttttgtttgt ctctgggcat ttcaaaggca actataaggc ccagctcaca cggttgaatc
     2941 acattacgaa ctgcaatggt gcggttttgt ccgtagagga actgctcatt ggtggagaaa
     3001 tgatcaaagc gggaactctg acactggaag aagtcagacg caagtttaac aatggcgaga
     3061 tcaatttccg ctcataatct agagcggccg cttcgagcag acatgataag atacattgat
     3121 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3181 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3241 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     3301 aacctctaca aatgtggtaa aatcgataag gatccaggtg gcacttttcg gggaaatgtg
     3361 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3421 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3481 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3541 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3601 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3661 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3721 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3781 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3841 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3901 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3961 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4021 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4081 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4141 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4201 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4261 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4321 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4381 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4621 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg acagatctgc
     5161 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     5221 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5281 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5341 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5401 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5461 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     5521 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgat
     5581 tcttctgaca caacagtctc gaacttaagc tgcagaagtt ggtcgtgagg cactgggcag
     5641 gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga
     5701 cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc
     5761 tttctctcca caggtgtcca
//
