LOCUS       sequence-214882-        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"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    570..665
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        905..1101
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      952..1087
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      1164..1173
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1170..1727
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     protein_bind    complement(1756..1855)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1885..1904)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1886..1904)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1909..1925)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1909..1925)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2027..2046)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2038..2847
                     /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(2105..2124)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2994..3582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3483..3502
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtataataa
      661 agttgggaga cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc
      721 tggaattcgt attaccgcca tgcattagtt attaatcgag atctggtgta aacacgccgt
      781 gggaaaaaat ttatggatct ggagtaacac gccgtgggaa aaaatttatg gatctggagt
      841 aacacgccgt gggaaaaaat ttatggatct ggagtaacac gccgtgggaa aaaatttatg
      901 gatctgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
      961 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     1021 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     1081 ggaggcctag gcttttgcaa aaagctggct agcgctaccg gactcagatc tcgagctcaa
     1141 gcttggtacc gagctcggat ccagccacca tgggagtcaa agttctgttt gccctgatct
     1201 gcatcgctgt ggccgaggcc aagcccaccg agaacaacga agacttcaac atcgtggccg
     1261 tggccagcaa cttcgcgacc acggatctcg atgctgaccg cgggaagttg cccggcaaga
     1321 agctgccgct ggaggtgctc aaagagatgg aagccaatgc ccggaaagct ggctgcacca
     1381 ggggctgtct gatctgcctg tcccacatca agtgcacgcc caagatgaag aagttcatcc
     1441 caggacgctg ccacacctac gaaggcgaca aagagtccgc acagggcggc ataggcgagg
     1501 cgatcgtcga cattcctgag attcctgggt tcaaggactt ggagcccatg gagcagttca
     1561 tcgcacaggt cgatctgtgt gtggactgca caactggctg cctcaaaggg cttgccaacg
     1621 tgcagtgttc tgacctgctc aagaagtggc tgccgcaacg ctgtgcgacc tttgccagca
     1681 agatccaggg ccaggtggac aagatcaagg gggccggtgg tgactaagcg gccgctcgag
     1741 catgcatcta gaggtaccca gctttcttgt acaaagttgg cattataaga aagcattgct
     1801 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     1861 cagctgcagg gcggccgcga tatcccctat agtgagtcgt attacatggt catagctgtt
     1921 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     1981 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     2041 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
     2101 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
     2161 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     2221 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     2281 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga
     2341 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     2401 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     2461 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     2521 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     2581 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     2641 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     2701 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     2761 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     2821 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag
     2881 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt
     2941 tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3001 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3061 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3121 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3181 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3241 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3301 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3361 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3421 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3481 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3541 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3601 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
