LOCUS       sequence-103221-        3957 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 3957)
  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 3957)
  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: pBDR
FEATURES             Location/Qualifiers
     source          1..3957
                     /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    161..194
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    complement(203..236)
                     /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      293..302
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             299..1729
                     /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    1856..1977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1893..1912)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1947..1966
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2093..2140)
                     /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(2141..2174)
                     /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(2258..2274)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2258..2274)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      2322..2353
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2354..2456
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2457..3272
                     /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(2530..2549)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3296..3323
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3335..3922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3415..3434)
                     /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 cgcggtgcgg gtgccagggc gtgcccttgg
      181 gctccccggg cgcgtactcc acataacttc gtatagcata cattatagca atttatcgaa
      241 gttcctattc cgaagttcct attctatcag aagtatagga acttcaccgg tcgccaccat
      301 ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg
      361 ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
      421 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
      481 cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg ccgacatccc
      541 cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
      601 ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca cgctgatcta
      661 caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac
      721 catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga
      781 gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagaccat
      841 ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca ccaagctgga
      901 catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg
      961 ccaccacctg ttcctggggc atggcaccgg cagcaccggc agcggcagct ccggcaccgc
     1021 ctcctccgag gacaacaaca tggccgtcat caaagagttc atgcgcttca aggtgcgcat
     1081 ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     1141 cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg
     1201 ggacatcctg tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc accccgccga
     1261 catccccgat tacaagaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     1321 cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg acggcacgct
     1381 gatctacaag gtgaagatgc gcggcaccaa cttccccccc gacggccccg taatgcagaa
     1441 gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa
     1501 gggcgagatc caccaggccc tgaagctgaa ggacggcggc cactacctgg tggagttcaa
     1561 gaccatctac atggccaaga agcccgtgca actgcccggc tactactacg tggacaccaa
     1621 gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg agcgctccga
     1681 gggccgccac cacctgttcc tgtacggcat ggacgagctg tacaagtaaa gcggccggcc
     1741 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     1801 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     1861 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     1921 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     1981 cgtaaattgt aagcgttaat gatccactag aggatccccg ggtaccgagc tcgaattcga
     2041 agttcctata ctcctataag aataggaact tcggaatagg aacttcatga tggaagttca
     2101 tatactttct agagaatagg aacttcggaa taggaacttc ataacttcgt ataggatact
     2161 ttatacgaag ttatcagctg ccgctcgagg cgcgtgatct tacggcatta tacgtatgat
     2221 cggtccacga tcagctagat tatctagtca gcttgatgtc atagctgttt cctgaggctc
     2281 aatactgacc atttaaatca tacctgacct ccatagcaga aagtcaaaag cctccgaccg
     2341 gaggcttttg acttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat
     2401 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatga
     2461 gccatattca acgggaaacg tcttgcttga agccgcgatt aaattccaac atggatgctg
     2521 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     2581 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     2641 ccaatgatgt tacagatgag atggtcaggc taaactggct gacggaattt atgcctcttc
     2701 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     2761 cagggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     2821 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     2881 acggcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     2941 gtgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     3001 tgcataaact cttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     3061 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     3121 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3181 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3241 agtttcactt gatgctcgat gagtttttct aatgaggacc taaatgtaat cacctggctc
     3301 accttcgggt gggcctttct gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3361 agcatcacaa aaatcgatgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3421 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3481 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3541 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3601 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3661 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3721 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3781 tatttggtat ctgcgctctg ctgaagccag ttacctcgga aaaagagttg gtagctcttg
     3841 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3901 gcgcagaaaa aaaggatctc aagaagatcc tttgattttc taccgaagaa aggccca
//
