LOCUS Exported 4959 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Single pathway reporter; hedgehog promoter-driven expression of mTurquoise2; MultiSite Gateway entry vector.. ACCESSION . VERSION . KEYWORDS pMuLE_ENTR_PTCH1-mTurquoise2_R4-R3 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4959 /organism="synthetic DNA construct" /mol_type="other DNA" 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 1453..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 4554..4581 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 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 //