LOCUS       sequence-214826-        4515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single pathway reporter; notch promoter-driven expression of 
            tdTomato; MultiSite Gateway entry vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4515)
  AUTHORS   Maier J, Elmenofi S, Taschauer A, Anton M, Sami H, Ogris M
  TITLE     Luminescent and fluorescent triple reporter plasmid constructs for 
            Wnt, Hedgehog and Notch pathway.
  JOURNAL   PLoS One. 2019 Dec 20;14(12):e0226570. doi: 
            10.1371/journal.pone.0226570. eCollection 2019.
  PUBMED    31860685
REFERENCE   2  (bases 1 to 4515)
  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: "PLoS One.";
            date: "2019-12-20"; volume: "14(12):e0226570. doi"; pages: " 
            10.1371/journal.pone.0226570. eCollection 2019"
COMMENT     Sequence Label: pMuLE_ENTR_CBF-tdTomato_L3-L2
FEATURES             Location/Qualifiers
     source          1..4515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(57..76)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(58..76)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(81..97)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(81..97)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(199..218)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             210..1019
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(277..296)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1166..1754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1655..1674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1908..1925
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2084..2111)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2203..2289)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2338..2360
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2352..2369
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2353..2369
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2386..2481
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2686..2882
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2733..2868
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      2958..2967
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2964..4394
                     /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
                     LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     primer_bind     complement(4415..4431)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4416..4432)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(join(4443..4515,1..27))
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tatcagtcaa aataaaatca ttatttgcca tccagctgca gggcggccgc gatatcccct
       61 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
      121 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
      181 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
      241 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
      301 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
      361 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
      421 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
      481 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
      541 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
      601 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
      661 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
      721 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
      781 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
      841 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
      901 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
      961 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     1021 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     1081 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     1141 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1201 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1261 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     1321 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1381 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1441 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1501 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1561 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1621 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1681 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1741 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     1801 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     1861 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     1921 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     1981 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2041 ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
     2101 gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
     2161 tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
     2221 agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg
     2281 ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc
     2341 agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gccccaaata atgattttat
     2401 tttgactgat agtgacctgt tcgttgcaac aaattgatga gcaatgcttt tttataatgc
     2461 caactttgta taataaagtt gctgtggaat gtgtgtccat atgataaccg tattaccgcc
     2521 atgcattagt tattaatcga gatctggtgt aaacacgccg tgggaaaaaa tttatggatc
     2581 tggagtaaca cgccgtggga aaaaatttat ggatctggag taacacgccg tgggaaaaaa
     2641 tttatggatc tggagtaaca cgccgtggga aaaaatttat ggatctgcat ctcaattagt
     2701 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2761 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2821 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2881 aaaagctggc tagcgctacc ggactcagat ctcgagctca agcttcgaat tcgccaccat
     2941 gccagagcca ggatcccgcc accatggtga gcaagggcga ggaggtcatc aaagagttca
     3001 tgcgcttcaa ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg
     3061 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg
     3121 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccaaggcgt
     3181 acgtgaagca ccccgccgac atccccgatt acaagaagct gtccttcccc gagggcttca
     3241 agtgggagcg cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc caggactcct
     3301 ccctgcagga cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg
     3361 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc
     3421 cccgcgacgg cgtgctgaag ggcgagatcc accaggccct gaagctgaag gacggcggcc
     3481 actacctggt ggagttcaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct
     3541 actactacgt ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg
     3601 aacagtacga gcgctccgag ggccgccacc acctgttcct ggggcatggc accggcagca
     3661 ccggcagcgg cagctccggc accgcctcct ccgaggacaa caacatggcc gtcatcaaag
     3721 agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag ttcgagatcg
     3781 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     3841 agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg tacggctcca
     3901 aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc ttccccgagg
     3961 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc gtgacccagg
     4021 actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc accaacttcc
     4081 cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc
     4141 tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag ctgaaggacg
     4201 gcggccgcta cctggtggag ttcaagacca tctacatggc caagaagccc gtgcaactgc
     4261 ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag gactacacca
     4321 tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgtac ggcatggacg
     4381 agctgtacaa gtaagaattc gatatcaagc ttatcgatac cgtcgacctc gagcctctag
     4441 atacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     4501 caacgaacag gtcac
//
