LOCUS       sequence-328797-        3644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single pathway reporter; notch promoter-driven expression of 
            secreted Gaussia luciferase; MultiSite Gateway entry vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3644)
  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 3644)
  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-GLuc_L3-L2
FEATURES             Location/Qualifiers
     source          1..3644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      210..219
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             216..773
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     protein_bind    complement(802..901)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(931..950)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(932..950)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(955..971)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(955..971)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1073..1092)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1084..1893
                     /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(1151..1170)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2040..2628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2529..2548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2782..2799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2958..2985)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3077..3163)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3212..3234
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3226..3243
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3227..3243
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3260..3355
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3595..3644,1..147)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      join(3642..3644,1..133)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
       61 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
      121 ttttttggag gcctaggctt ttgcaaaaag ctggctagcg ctaccggact cagatctcga
      181 gctcaagctt ggtaccgagc tcggatccag ccaccatggg agtcaaagtt ctgtttgccc
      241 tgatctgcat cgctgtggcc gaggccaagc ccaccgagaa caacgaagac ttcaacatcg
      301 tggccgtggc cagcaacttc gcgaccacgg atctcgatgc tgaccgcggg aagttgcccg
      361 gcaagaagct gccgctggag gtgctcaaag agatggaagc caatgcccgg aaagctggct
      421 gcaccagggg ctgtctgatc tgcctgtccc acatcaagtg cacgcccaag atgaagaagt
      481 tcatcccagg acgctgccac acctacgaag gcgacaaaga gtccgcacag ggcggcatag
      541 gcgaggcgat cgtcgacatt cctgagattc ctgggttcaa ggacttggag cccatggagc
      601 agttcatcgc acaggtcgat ctgtgtgtgg actgcacaac tggctgcctc aaagggcttg
      661 ccaacgtgca gtgttctgac ctgctcaaga agtggctgcc gcaacgctgt gcgacctttg
      721 ccagcaagat ccagggccag gtggacaaga tcaagggggc cggtggtgac taagcggccg
      781 ctcgagcatg catctagagg tacccagctt tcttgtacaa agttggcatt ataagaaagc
      841 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
      901 gccatccagc tgcagggcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata
      961 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     1021 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     1081 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     1141 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1201 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1261 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1321 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1381 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1441 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1501 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     1561 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1621 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     1681 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     1741 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     1801 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     1861 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     1921 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     1981 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2041 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2101 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2161 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2221 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2281 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2341 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2401 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2461 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2521 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2581 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2641 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2701 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2761 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     2821 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2881 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     2941 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     3001 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     3061 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     3121 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3181 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     3241 agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac ctgttcgttg
     3301 caacaaattg atgagcaatg cttttttata atgccaactt tgtataataa agttgggaga
     3361 cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgt
     3421 attaccgcca tgcattagtt attaatcgag atctggtgta aacacgccgt gggaaaaaat
     3481 ttatggatct ggagtaacac gccgtgggaa aaaatttatg gatctggagt aacacgccgt
     3541 gggaaaaaat ttatggatct ggagtaacac gccgtgggaa aaaatttatg gatctgcatc
     3601 tcaattagtc agcaaccata gtcccgcccc taactccgcc catc
//
