LOCUS       sequence-103226-        4086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for tdTomato knock-in via landing pad.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4086)
  AUTHORS   Ohtsuka M, Miura H, Mochida K, Hirose M, Hasegawa A, Ogura A, 
            Mizutani R, Kimura M, Isotani A, Ikawa M, Sato M, Gurumurthy CB
  TITLE     One-step generation of multiple transgenic mouse lines using an 
            improved Pronuclear Injection-based Targeted Transgenesis (i-PITT).
  JOURNAL   BMC Genomics. 2015 Apr 9;16:274. doi: 10.1186/s12864-015-1432-5.
  PUBMED    25887549
REFERENCE   2  (bases 1 to 4086)
  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: "BMC 
            Genomics."; date: "2015-04-9"; volume: "16:274. doi"; pages: " 
            10.1186/s12864-015-1432-5"
COMMENT     Sequence Label: pBFB
FEATURES             Location/Qualifiers
     source          1..4086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..22
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6..22
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(158..191)
                     /label=loxJTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(192..225)
                     /label=loxJTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(226..259)
                     /label=loxJTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(260..293)
                     /label=loxJTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(294..327)
                     /label=loxJTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(328..361)
                     /label=loxJTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      422..431
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             428..1858
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    1985..2106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2022..2041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2076..2095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2222..2269)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    complement(2270..2303)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(2387..2403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2387..2403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      2451..2482
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2483..2585
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2586..3401
                     /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="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2659..2678)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3425..3452
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3464..4051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3544..3563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccgtgtaaa acgacggcca gtttatctag tcagcttgat tctagctgat cgtggaccgg
       61 aaggtgagcc agtgagttga ttgcagtcca gttacgctgg agtctgaggc tcgtcctgaa
      121 tgatatgcgg cctcggatcc gcgatcgatc cgctagcata acttcgtata gcatacatta
      181 tagcaattta tataacttcg tatagcatac attatagcaa tttatataac ttcgtatagc
      241 atacattata gcaatttata taacttcgta tagcatacat tatagcaatt tatataactt
      301 cgtatagcat acattatagc aatttatata acttcgtata gcatacatta tagcaattta
      361 tcctaggaag ttcctattcc gaagttccta ttctatcaga agtataggaa cttcaccggt
      421 cgccaccatg gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg
      481 catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
      541 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
      601 ctgggacatc ctgtcccccc agttcatgta cggctccaag gcgtacgtga agcaccccgc
      661 cgacatcccc gattacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
      721 gaacttcgag gacggcggtc tggtgaccgt gacccaggac tcctccctgc aggacggcac
      781 gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca
      841 gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct
      901 gaagggcgag atccaccagg ccctgaagct gaaggacggc ggccactacc tggtggagtt
      961 caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact acgtggacac
     1021 caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc
     1081 cgagggccgc caccacctgt tcctggggca tggcaccggc agcaccggca gcggcagctc
     1141 cggcaccgcc tcctccgagg acaacaacat ggccgtcatc aaagagttca tgcgcttcaa
     1201 ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     1261 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc
     1321 cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccaaggcgt acgtgaagca
     1381 ccccgccgac atccccgatt acaagaagct gtccttcccc gagggcttca agtgggagcg
     1441 cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc caggactcct ccctgcagga
     1501 cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt
     1561 aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg
     1621 cgtgctgaag ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt
     1681 ggagttcaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct actactacgt
     1741 ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     1801 gcgctccgag ggccgccacc acctgttcct gtacggcatg gacgagctgt acaagtaaag
     1861 cggccggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     1921 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     1981 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2041 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2101 atcttaaggc gtaaattgta agcgttaatg atccactaga ggatccccgg gtaccgagct
     2161 cgaattcgaa gttcctatac tcctataaga ataggaactt cggaatagga acttcatgat
     2221 ggaagttcat atactttcta gagaatagga acttcggaat aggaacttca taacttcgta
     2281 taggatactt tatacgaagt tatcagctgc cgctcgaggc gcgtgatctt acggcattat
     2341 acgtatgatc ggtccacgat cagctagatt atctagtcag cttgatgtca tagctgtttc
     2401 ctgaggctca atactgacca tttaaatcat acctgacctc catagcagaa agtcaaaagc
     2461 ctccgaccgg aggcttttga cttgatcggc acgtaagagg ttccaacttt caccataatg
     2521 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag
     2581 ctaaaatgag ccatattcaa cgggaaacgt cttgcttgaa gccgcgatta aattccaaca
     2641 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     2701 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     2761 gtagcgttgc caatgatgtt acagatgaga tggtcaggct aaactggctg acggaattta
     2821 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     2881 ctgcgatccc agggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     2941 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     3001 gtccttttaa cggcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     3061 gtttggttgg tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     3121 ggaaagaaat gcataaactc ttgccattct caccggattc agtcgtcact catggtgatt
     3181 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     3241 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     3301 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     3361 ataaattgca gtttcacttg atgctcgatg agtttttcta atgaggacct aaatgtaatc
     3421 acctggctca ccttcgggtg ggcctttctg cgttgctggc gtttttccat aggctccgcc
     3481 cccctgacga gcatcacaaa aatcgatgct caagtcagag gtggcgaaac ccgacaggac
     3541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3901 gaagaacagt atttggtatc tgcgctctgc tgaagccagt tacctcggaa aaagagttgg
     3961 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4021 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgattttct accgaagaaa
     4081 ggccca
//
