LOCUS       sequence-214856-        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"
     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    complement(603..727)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(2126..2128,2129..2131,2132..2839)
                     /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(2171..2192)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2432..2451)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2779..2800
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    3022..3145
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3200..3219)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3201..3219)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3224..3240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3224..3240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3342..3361)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3353..4162
                     /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(3420..3439)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4309..4897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4798..4817
                     /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 tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttgggag acccaagctt ggtaccgagc tcttacgcgt gctagagctt gcatgcctgc
      781 aggtcgacac actggcgcac tatccagcac gctgtcatta ttggtgcagg cctgggaatg
      841 cgagccggac ctcccagtat ttgctgcagc ttgaggccaa ggtgcctctg aaaggctttt
      901 ctttgaggaa tttcagaaat tgtttaagcg cttaaaaaaa tattacattt ccccccgtgt
      961 ctaccgcgag gcaaatgttc gtgtgtgtgc gtgcgcgcgc gtgtgtgcgc gcaaaggcat
     1021 cccacaaaaa gaagacactg ttgaaaagaa aaggaataag agagaaagga gggggagcat
     1081 tccttaatgg aagtattgca tgcgagaagg ttggcgctct cagaggctgc cccgaagccg
     1141 aggatgcaca cactgggttg cctacctggg tggtctctct actttggtga gctgttgcta
     1201 agcagctaat aatagtcatg tttgctgagt aatttctgcc ctccgcgagc caatcgcgtc
     1261 gcagaaaccc aagccatctg acagccccgg gggcgggagc tccagccttt ttctctttcg
     1321 ctcgctctct ccccctccac cccctcccct cccctcctcc tcttctttct ccaaatggag
     1381 agagttcttt tttttcttct tccatctcat ctattgaatc aaggagctgc tgcggccgct
     1441 gcccgctgca cacacacaga gcggagtccc caggtcccgg gagcgagaga ggcgcgctgc
     1501 acggggacag aatggtccag cgggttgccg aggagcacaa gaaagcagag tccgggaccg
     1561 agcagccacc gcgaacccag cagccagagc ccgagcagcc cgagcagcag ctcctggagc
     1621 cgccaccgcc agcagcagcc accgcgggag cagcggcagc tgcggctgcc cgggcccggc
     1681 agcgctgaga cccgcagggc accctcggac cccgcggcgg cggcggctct gcgcttgccc
     1741 tccgcggccg ctcgggcgac ccgggaggcg ccgagagagc cagcgcggcc ggcgggagca
     1801 gcggggattc gctggctctt tctgcagtga aggggtcgcg gcgcggggcg gggggcgggt
     1861 agggggagtt ggggaccgca aggagtgccg cggaagcgcc cgaaggacag gctcgctcgg
     1921 cgcgccggct ctcgctcttc cgcgaactgg atgtgggcag cggcggccgc agagacctcg
     1981 ggacccccgc gcaatgtggc aatggaaggc gcagggtctg actccccggc agcggccgcg
     2041 gccgcagcgg cagcagcgcc cgccgtgtga gcagcagcag cggctggtct gtcaaccgga
     2101 gcggatctgc gatctaagta agcttatggt gagcaagggc gaggagctgt tcaccggggt
     2161 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     2221 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     2281 caagctgccc gtgccctggc ccaccctcgt gaccaccctg tcctggggcg tgcagtgctt
     2341 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2401 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2461 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2521 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tactttagcg acaacgtcta
     2581 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     2641 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2701 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     2761 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2821 cggcatggac gagctgtaca gtgagctgca gaacaagctg gaagagttgg atctggactc
     2881 gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggat
     2941 ccactagtaa cggccgccag tgtgctggaa ttctgcagat atccatcaca ctggcggccg
     3001 ctcgagcatg catctagagg acaactttgt ataataaagt tgaacgagaa acgtaaaatg
     3061 atataaatat caatatatta aattagattt tgcataaaaa acagactaca taatactgta
     3121 aaacacaaca tatccagtca ctatgaatca actacttaga tggtattagt gacctgtact
     3181 gcagggcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     3241 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     3301 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     3361 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     3421 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     3481 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3541 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3601 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3661 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3721 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3781 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3841 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3901 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3961 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     4021 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     4081 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     4141 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     4201 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     4261 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4321 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4381 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4441 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4501 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4561 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4621 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4681 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4741 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4801 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4861 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4921 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
