LOCUS       sequence-124428-        7594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEN expression construct for targeting the murine ROSA26 locus. It
            recognize the sequence: CTGCAACTCCAGTCT. Can be used with the pBS SK
            mCherryROSAbsr plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7594)
  TITLE     rosa26 knock-in plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7594)
  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            "Unpublished"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: pCAGGS-ROSAf
FEATURES             Location/Qualifiers
     source          1..7594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1726..1735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1747..1767
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1777..1800
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1801..1827
                     /codon_start=1
                     /product="epitope tag from the baculovirus Autographa 
                     californica GP64 envelope fusion protein"
                     /label=AcV5 tag
                     /translation="SWKDASGWS"
     CDS             3934..4521
                     /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
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     primer_bind     complement(4618..4637)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4683..4738
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4737..4756)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5097..5113)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5097..5113)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5110..5132)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5121..5137
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5145..5151,5152..5169,5170..5175))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5190..5211
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5270..5465
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5316..5451
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5471..5605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5508..5527)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5562..5581
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5673..5690)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5844..6432)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5924..5943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6603..7394,7395..7463))
                     /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     7226..7245
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7464..7568)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attccgccac catggcttcc
     1741 tcccctccaa agaaaaagag aaaggttgcg gccgctgact acaaggatga cgacgataaa
     1801 agttggaagg acgcaagtgg ttggtctaga atgcatgcgg ccccgcgacg gcgtgctgcg
     1861 caaccctccg acgcttcgcc ggccgcgcag gtggatctac gcacgctcgg ctacagtcag
     1921 cagcagcaag agaagatcaa accgaaggtg cgttcgacag tggcgcagca ccacgaggca
     1981 ctggtgggcc atgggtttac acacgcgcac atcgttgcgc tcagccaaca cccggcagcg
     2041 ttagggaccg tcgctgtcac gtatcagcac ataatcacgg cgttgccaga ggcgacacac
     2101 gaagacatcg ttggcgtcgg caaacagtgg tccggcgcac gcgccctgga ggccttgctc
     2161 acggatgcgg gggagttgag aggtccgccg ttacagttgg acacaggcca acttgtgaag
     2221 attgcaaaac gtggcggcgt gaccgcaatg gaggcagtgc atgcatcgcg caatgcactg
     2281 acgggtgccc ccctgaacct gaccccggac caagtggtgg ctatcgccag ccacgatggc
     2341 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
     2401 ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa
     2461 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg
     2521 gtggctatcg ccagcaacaa tggcggcaag caagcgctcg aaacggtgca gcggctgttg
     2581 ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac
     2641 gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac
     2701 catggcctga ccccggacca agtggtggct atcgccagca acattggcgg caagcaagcg
     2761 ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac
     2821 caagtggtgg ctatcgccag caacattggc ggcaagcaag cgctcgaaac ggtgcagcgg
     2881 ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc
     2941 agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc
     3001 caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag
     3061 caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact
     3121 ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg
     3181 cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct
     3241 atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg
     3301 ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacattggc
     3361 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
     3421 ctgaccccgg accaagtggt ggctatcgcc agcaacaatg gcggcaagca agcgctcgaa
     3481 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg
     3541 gtggctatcg ccagcaacgg tggcggcaag caagcgctcg aaacggtgca gcggctgttg
     3601 ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac
     3661 gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac
     3721 catggcctga ccccggacca agtggtggct atcgccagca acggtggcgg caagcaagcg
     3781 ctcgaaagca ttgtggccca gctgagccgg cctgatccgg cgttggccgc gttgaccaac
     3841 gaccacctcg tcgccttggc ctgcctcggc ggacgtcctg ccatggatgc agtgaaaaag
     3901 ggattgccgc acgcgccgga attgatcaga tcccagctag tcaaaagtga actggaggag
     3961 aagaaatctg aacttcgtca taaattgaaa tatgtgcctc atgaatatat tgaattaatt
     4021 gaaattgcca gaaattccac tcaggataga attcttgaaa tgaaggtaat ggaatttttt
     4081 atgaaagttt atggatatag aggtaaacat ttgggtggat caaggaaacc ggacggagca
     4141 atttatactg tcggatctcc tattgattac ggtgtgatcg tggatactaa agcttatagc
     4201 ggaggttata atctgccaat tggccaagca gatgaaatgc aacgatatgt cgaagaaaat
     4261 caaacacgaa acaaacatat caaccctaat gaatggtgga aagtctatcc atcttctgta
     4321 acggaattta agtttttatt tgtgagtggt cactttaaag gaaactacaa agctcagctt
     4381 acacgattaa atcatatcac taattgtaat ggagctgttc ttagtgtaga agagctttta
     4441 attggtggag aaatgattaa agccggcaca ttaaccttag aggaagtgag aaggaaattt
     4501 aataacggcg agatcaactt cgcggccgac tgaggaattc actcctcagg tgcaggctgc
     4561 ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct
     4621 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     4681 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     4741 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     4801 tttggcaaca tatgccatat gctggctgcc atgaacaaag gtggctataa agaggtcatc
     4861 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     4921 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     4981 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     5041 tccctcttct cttatgaaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
     5101 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5161 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5221 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     5281 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5341 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5401 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5461 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5521 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5581 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     5641 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5701 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5761 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5821 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5881 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5941 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6001 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     6061 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6121 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6181 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6241 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6301 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6361 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6421 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6481 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6541 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6601 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6661 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6721 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6781 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6841 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6901 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6961 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7021 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7081 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7141 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7201 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7261 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7321 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7381 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7441 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7501 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7561 gttccgcgca catttccccg aaaagtgcca cctg
//
