LOCUS       sequence-221594-        3866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single pathway reporter; wnt promoter-driven expression of iRFP; 
            MultiSite Gateway entry vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3866)
  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 3866)
  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_TOP-iRFP_L5-L4
FEATURES             Location/Qualifiers
     source          1..3866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        55..86
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             160..1110
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     protein_bind    complement(1196..1291)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1322..1341)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1323..1341)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1346..1362)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1346..1362)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1464..1483)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1475..2284
                     /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(1542..1561)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2431..3019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2920..2939
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3173..3190
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3349..3376)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3468..3554)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3603..3625
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3617..3634
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3618..3634
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3651..3746
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggtaagatca aaggggcgcg cccgcgtgct agcccgggct cgagatctag actctagagg
       61 gtatataatg gaagctcgaa ttccagcttg gcattccggt actgttggta aaaagcttgg
      121 cattccggta ctgttggtaa agccaccatc agctgcacca tggcggaagg atccgtcgcc
      181 aggcagcctg acctcttgac ctgcgacgat gagccgatcc atatccccgg tgccatccaa
      241 ccgcatggac tgctgctcgc cctcgccgcc gacatgacga tcgttgccgg cagcgacaac
      301 cttcccgaac tcaccggact ggcgatcggc gccctgatcg gccgctctgc ggccgatgtc
      361 ttcgactcgg agacgcacaa ccgtctgacg atcgccttgg ccgagcccgg ggcggccgtc
      421 ggagcaccga tcactgtcgg cttcacgatg cgaaaggacg caggcttcat cggctcctgg
      481 catcgccatg atcagctcat cttcctcgag ctcgagcctc cccagcggga cgtcgccgag
      541 ccgcaggcgt tcttccgccg caccaacagc gccatccgcc gcctgcaggc cgccgaaacc
      601 ttggaaagcg cctgcgccgc cgcggcgcaa gaggtgcgga agattaccgg cttcgatcgg
      661 gtgatgatct atcgcttcgc ctccgacttc agcggcgaag tgatcgcaga ggatcggtgc
      721 gccgaggtcg agtcaaaact aggcctgcac tatcctgcct caaccgtgcc ggcgcaggcc
      781 cgtcggctct ataccatcaa cccggtacgg atcattcccg atatcaatta tcggccggtg
      841 ccggtcaccc cagacctcaa tccggtcacc gggcggccga ttgatcttag cttcgccatc
      901 ctgcgcagcg tctcgcccgt ccatctggaa ttcatgcgca acataggcat gcacggcacg
      961 atgtcgatct cgattttgcg cggcgagcga ctgtggggat tgatcgtttg ccatcaccga
     1021 acgccgtact acgtcgatct cgatggccgc caagcctgcg agctagtcgc ccaggttctg
     1081 gcctggcaga tcggcgtgat ggaagagtga gtcactagta acggccgcca gtgtgctgga
     1141 attctgcaga tatccatcac actggcggcc gctcgagcat gcatctagag gcacccaact
     1201 tttctataca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1261 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagggcg gccgcgatat
     1321 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1381 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1441 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     1501 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1561 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     1621 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1681 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     1741 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     1801 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     1861 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     1921 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     1981 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2041 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2101 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2161 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2221 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2281 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     2341 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     2401 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2461 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2521 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2581 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2641 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2701 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2761 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2821 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2881 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2941 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3001 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3061 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3121 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3181 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     3241 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     3301 cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc
     3361 gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc
     3421 caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc
     3481 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc
     3541 tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt
     3601 ttcccagtca cgacgttgta aaacgacggc cagtcttaag ctcgggcccc aaataatgat
     3661 tttattttga ctgatagtga cctgttcgtt gcaacaaatt gatgagcaat gcttttttat
     3721 aatgccaact ttgtatacaa aagttgggag acccaagctt ggtaccgagc tcttacgcga
     3781 gatcaaaggg ggtaagatca aagggggtaa gatcaaaggg gcgcgagatc aaagggggta
     3841 agatcaaagg gggtaagatc aaaggg
//
