LOCUS       sequence-221601-        4959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single pathway reporter; hedgehog promoter-driven expression of 
            mTurquoise2; MultiSite Gateway entry vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4959)
  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 4959)
  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_PTCH1-mTurquoise2_R4-R3
FEATURES             Location/Qualifiers
     source          1..4959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(33..53)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             join(1453..1455,1456..1458,1459..2166)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(1498..1519)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1759..1778)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2106..2127
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    2349..2472
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2527..2546)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2528..2546)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2551..2567)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2551..2567)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2669..2688)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2680..3489
                     /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(2747..2766)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3636..4224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4125..4144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4378..4395
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4554..4581)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4673..4759)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4808..4830
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4822..4839
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4823..4839
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 attgatattt atatcatttt acgtttctcg ttcaactttt ctatacaaag ttgggagacc
       61 caagcttggt accgagctct tacgcgtgct agagcttgca tgcctgcagg tcgacacact
      121 ggcgcactat ccagcacgct gtcattattg gtgcaggcct gggaatgcga gccggacctc
      181 ccagtatttg ctgcagcttg aggccaaggt gcctctgaaa ggcttttctt tgaggaattt
      241 cagaaattgt ttaagcgctt aaaaaaatat tacatttccc cccgtgtcta ccgcgaggca
      301 aatgttcgtg tgtgtgcgtg cgcgcgcgtg tgtgcgcgca aaggcatccc acaaaaagaa
      361 gacactgttg aaaagaaaag gaataagaga gaaaggaggg ggagcattcc ttaatggaag
      421 tattgcatgc gagaaggttg gcgctctcag aggctgcccc gaagccgagg atgcacacac
      481 tgggttgcct acctgggtgg tctctctact ttggtgagct gttgctaagc agctaataat
      541 agtcatgttt gctgagtaat ttctgccctc cgcgagccaa tcgcgtcgca gaaacccaag
      601 ccatctgaca gccccggggg cgggagctcc agcctttttc tctttcgctc gctctctccc
      661 cctccacccc ctcccctccc ctcctcctct tctttctcca aatggagaga gttctttttt
      721 ttcttcttcc atctcatcta ttgaatcaag gagctgctgc ggccgctgcc cgctgcacac
      781 acacagagcg gagtccccag gtcccgggag cgagagaggc gcgctgcacg gggacagaat
      841 ggtccagcgg gttgccgagg agcacaagaa agcagagtcc gggaccgagc agccaccgcg
      901 aacccagcag ccagagcccg agcagcccga gcagcagctc ctggagccgc caccgccagc
      961 agcagccacc gcgggagcag cggcagctgc ggctgcccgg gcccggcagc gctgagaccc
     1021 gcagggcacc ctcggacccc gcggcggcgg cggctctgcg cttgccctcc gcggccgctc
     1081 gggcgacccg ggaggcgccg agagagccag cgcggccggc gggagcagcg gggattcgct
     1141 ggctctttct gcagtgaagg ggtcgcggcg cggggcgggg ggcgggtagg gggagttggg
     1201 gaccgcaagg agtgccgcgg aagcgcccga aggacaggct cgctcggcgc gccggctctc
     1261 gctcttccgc gaactggatg tgggcagcgg cggccgcaga gacctcggga cccccgcgca
     1321 atgtggcaat ggaaggcgca gggtctgact ccccggcagc ggccgcggcc gcagcggcag
     1381 cagcgcccgc cgtgtgagca gcagcagcgg ctggtctgtc aaccggagcg gatctgcgat
     1441 ctaagtaagc ttatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1501 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1561 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1621 ccctggccca ccctcgtgac caccctgtcc tggggcgtgc agtgcttcgc ccgctacccc
     1681 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1741 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1801 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1861 atcctggggc acaagctgga gtacaactac tttagcgaca acgtctatat caccgccgac
     1921 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1981 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2041 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     2101 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2161 ctgtacagtg agctgcagaa caagctggaa gagttggatc tggactcgta caagtaaagc
     2221 ggccgcgact ctagatcata atcagccata ccacatttgt agaggatcca ctagtaacgg
     2281 ccgccagtgt gctggaattc tgcagatatc catcacactg gcggccgctc gagcatgcat
     2341 ctagaggaca actttgtata ataaagttga acgagaaacg taaaatgata taaatatcaa
     2401 tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat
     2461 ccagtcacta tgaatcaact acttagatgg tattagtgac ctgtactgca gggcggccgc
     2521 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     2581 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     2641 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     2701 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     2761 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     2821 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2881 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     2941 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     3001 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     3061 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     3121 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     3181 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     3241 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     3301 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     3361 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     3421 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     3481 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     3541 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     3601 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3661 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3721 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3781 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3841 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3901 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3961 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4021 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4081 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4141 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4201 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4261 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4321 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     4381 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     4441 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     4501 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc
     4561 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg
     4621 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc
     4681 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact
     4741 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg
     4801 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gcccctacag
     4861 gtcactaata ccatctaagt agttgattca tagtgactgg atatgttgtg ttttacagta
     4921 ttatgtagtc tgttttttat gcaaaatcta atttaatat
//
