Gpx1 fw CTAGCGTTAATTAAAGTGCTGCTGTCTCGGGGGGGTTT
3
Gpx1 fw CTAGCGTTAATTAAAGTGCTGCTGTCTCGGGGGGGTTT Gpx1 rev CTAGCGGCGGCCGCACCCATCTCGAGGTGGTATTTTCTGT Gpx2 fw CTAGCGTTAATTAAAGACTTGGGTAAGCTCTGGGCC Gpx2 rev CTAGCGGCGGCCGCTTCTAGCAGAGTGGCTCCAGGC Gpx3 fw CTAGCGTTAATTAACACGGACCCCATGGCAGGGGTGGC Gpx3 rev CTAGCGGCGGCCGCATGCACTAAGGGCTGGCCCCTCAG Gpx4 fw CTAGCGTTAATTAACCACGCCCTCGGAGCCTTCCACCG Gpx4 rev CTAGCGGCGGCCGCGACCTTCCTCCGGCGGGGTGCACG Gpx6 fw CTAGCGTTAATTAACCCACCTCACATGAAGGGAAGG Gpx6 rev CTAGCGGCGGCCGCTGCACTAAGGGACAGGCACCAAGG Dio1 fw CTAGCGTTAATTAATTTTAACTCTGTGTCTTTACATATTTG Dio1 rev CTAGCGGCGGCCGCAGTTTACTGCATCTTATAACATTTTCT Dio2 fw CTAGCGTTAATTAACAGAGATGTGCAGAGTTGACCAGT Dio2 rev CTAGCGGCGGCCGCGAGAGGCAAACTAATGTGACTACATCC Dio3 fw CTAGCGTTAATTAATGTTTTGGGTGCACAGGAGCCCCA Dio3 rev CTAGCGGCGGCCGCTTTAGGTGCTGCTTTGAGGCCCCT Trxr1 fw CTAGCGTTAATTAACATTTGGCAGGGCATCGAAGGGAT Trxr1 rev CTAGCGGCGGCCGCAGGTCATGCTAAAACTGATTTGACAG Trxr2 fw CTAGCGTTAATTAACACCCCCCCCCAGGCTCCTGGTGC Trxr2 rev CTAGCGGCGGCCGCTATTTGCATTGCAGAAATGCCAGGGG Trxr3 fw CTAGCGTTAATTAACTGACAGCGAGAAGCAGTGGGACT Trxr3 rev CTAGCGGCGGCCGCGGGAAGCTCAGTCCTGGCTTGCGA SelH fw CTAGCGTTAATTAACTTTGTGTCCCTGGTGATGTTGGA SelH rev CTAGCGGCGGCCGCTGAACCTTCATTTCTGGCAGGGAA SelI fw CTAGCGTTAATTAATATTTTTCACTGAATGAAGTTTGTGC SelI rev CTAGCGGCGGCCGCATCAACACAGTGGTGCCTATTTTAT SelK fw CTAGCGTTAATTAAACAAGGACTGCTCTGTGTCCTCAC SelK rev CTAGCGGCGGCCGCAACATCTGCATTTATGGAACTGCAT SelM fw CTAGCGTTAATTAAGACCTACCTGCCTGAGTCCTGGAGACAGAA SelM rev CTAGCGGCGGCCGCCCCATCCCTGCTGGCCCGGGG SelN fw CTAGCGTTAATTAAAGGGGCAGTGGCTTCCCCGGCAGC SelN rev CTAGCGGCGGCCGCCTACTTCTTCCCATAGGTGCAGCTGC SelO fw CTAGCGTTAATTAATGCCCTGGCCCATGCACACCCGTC SelO rev CTAGCGGCGGCCGCGGGACCAGGCTGCACTGCTGAGCC SelP1 fw CTAGCGTTAATTAATTTTCTTTTTCCAGTGTTCTATTTGC SelP1 rev CTAGCGGCGGCCGCGTTCTTCCTCCATTCTAAACTGCTAA SelP2 fw CTAGCGTTAATTAATATATTGCTTAGTAAGTATTTCCATAGTC SelP2 rev CTAGCGGCGGCCGCCATTCTTGCTTAATAGTATTAACCATAAA SelS fw CTAGCGTTAATTAATAGGACAGTCTCTGTGACAGGTTGCG SelS rev CTAGCGGCGGCCGCGAAACAAACCCCATCAACTGTCCAC SelT fw CTAGCGTTAATTAAGATCATTGCAAGAGCAGCGTGACT SelT rev CTAGCGGCGGCCGCGAGGTACAACGAGCCTGCCAAGAAA SelV fw CTAGCGTTAATTAAGCAAGGGGTGGAGCTGGAGGAGTC SelV rev CTAGCGGCGGCCGCTGAGCTCACCCCAGCCCCAGCCAC SelW fw CTAGCGTTAATTAACTTGACCCAGCCCCTCTCAGCAGA SelW rev CTAGCGGCGGCCGCGCCCCCAACAGCAGCCACGAGAACA SelX fw CTAGCGTTAATTAACCTGCCAGCCGCCCTGGCCCT SelX rev CTAGCGGCGGCCGCGCCTGCCTTCCCAGCAGGTCACAGA Sel15 fw CTAGCGTTAATTAAAGAGTGAAACATTCACAAAGATTTGCG Sel15 rev CTAGCGGCGGCCGCAAAACTCAAAAATTTGCCAAGTATGT SPS2 fw CTAGCGTTAATTAAACCTGCAACCATCTGACTTGGTCTCTGT SPS2 rev CTAGCGGCGGCCGCCACTGAGGCCTGGGCTCTGAGGCT Supplementary Figure 1. List of primers used for PCR amplification of human SECIS elements and cloning downstream of luciferase UGA 258 coding sequence.
description
Gpx1 fw CTAGCGTTAATTAAAGTGCTGCTGTCTCGGGGGGGTTT Gpx1 rev CTAGCGGCGGCCGCACCCATCTCGAGGTGGTATTTTCTGT Gpx2 fw CTAGCGTTAATTAAAGACTTGGGTAAGCTCTGGGCC Gpx2 rev CTAGCGGCGGCCGCTTCTAGCAGAGTGGCTCCAGGC Gpx3 fw CTAGCGTTAATTAACACGGACCCCATGGCAGGGGTGGC - PowerPoint PPT Presentation
Transcript of Gpx1 fw CTAGCGTTAATTAAAGTGCTGCTGTCTCGGGGGGGTTT
Gpx1 fw CTAGCGTTAATTAAAGTGCTGCTGTCTCGGGGGGGTTTGpx1 rev CTAGCGGCGGCCGCACCCATCTCGAGGTGGTATTTTCTGT Gpx2 fw CTAGCGTTAATTAAAGACTTGGGTAAGCTCTGGGCC Gpx2 rev CTAGCGGCGGCCGCTTCTAGCAGAGTGGCTCCAGGC Gpx3 fw CTAGCGTTAATTAACACGGACCCCATGGCAGGGGTGGCGpx3 rev CTAGCGGCGGCCGCATGCACTAAGGGCTGGCCCCTCAG Gpx4 fw CTAGCGTTAATTAACCACGCCCTCGGAGCCTTCCACCGGpx4 rev CTAGCGGCGGCCGCGACCTTCCTCCGGCGGGGTGCACGGpx6 fw CTAGCGTTAATTAACCCACCTCACATGAAGGGAAGG Gpx6 rev CTAGCGGCGGCCGCTGCACTAAGGGACAGGCACCAAGGDio1 fw CTAGCGTTAATTAATTTTAACTCTGTGTCTTTACATATTTG
Dio1 rev CTAGCGGCGGCCGCAGTTTACTGCATCTTATAACATTTTCTDio2 fw CTAGCGTTAATTAACAGAGATGTGCAGAGTTGACCAGT Dio2 rev CTAGCGGCGGCCGCGAGAGGCAAACTAATGTGACTACATCC Dio3 fw CTAGCGTTAATTAATGTTTTGGGTGCACAGGAGCCCCA Dio3 rev CTAGCGGCGGCCGCTTTAGGTGCTGCTTTGAGGCCCCT Trxr1 fw CTAGCGTTAATTAACATTTGGCAGGGCATCGAAGGGAT Trxr1 rev CTAGCGGCGGCCGCAGGTCATGCTAAAACTGATTTGACAGTrxr2 fw CTAGCGTTAATTAACACCCCCCCCCAGGCTCCTGGTGC Trxr2 rev CTAGCGGCGGCCGCTATTTGCATTGCAGAAATGCCAGGGG Trxr3 fw CTAGCGTTAATTAACTGACAGCGAGAAGCAGTGGGACT Trxr3 rev CTAGCGGCGGCCGCGGGAAGCTCAGTCCTGGCTTGCGA SelH fw CTAGCGTTAATTAACTTTGTGTCCCTGGTGATGTTGGA SelH rev CTAGCGGCGGCCGCTGAACCTTCATTTCTGGCAGGGAA SelI fw CTAGCGTTAATTAATATTTTTCACTGAATGAAGTTTGTGC SelI rev CTAGCGGCGGCCGCATCAACACAGTGGTGCCTATTTTAT SelK fw CTAGCGTTAATTAAACAAGGACTGCTCTGTGTCCTCAC SelK rev CTAGCGGCGGCCGCAACATCTGCATTTATGGAACTGCAT SelM fw CTAGCGTTAATTAAGACCTACCTGCCTGAGTCCTGGAGACAGAASelM rev CTAGCGGCGGCCGCCCCATCCCTGCTGGCCCGGGGSelN fw CTAGCGTTAATTAAAGGGGCAGTGGCTTCCCCGGCAGC SelN rev CTAGCGGCGGCCGCCTACTTCTTCCCATAGGTGCAGCTGC SelO fw CTAGCGTTAATTAATGCCCTGGCCCATGCACACCCGTC SelO rev CTAGCGGCGGCCGCGGGACCAGGCTGCACTGCTGAGCC SelP1 fw CTAGCGTTAATTAATTTTCTTTTTCCAGTGTTCTATTTGCSelP1 rev CTAGCGGCGGCCGCGTTCTTCCTCCATTCTAAACTGCTAASelP2 fw CTAGCGTTAATTAATATATTGCTTAGTAAGTATTTCCATAGTCSelP2 rev CTAGCGGCGGCCGCCATTCTTGCTTAATAGTATTAACCATAAASelS fw CTAGCGTTAATTAATAGGACAGTCTCTGTGACAGGTTGCGSelS rev CTAGCGGCGGCCGCGAAACAAACCCCATCAACTGTCCACSelT fw CTAGCGTTAATTAAGATCATTGCAAGAGCAGCGTGACTSelT rev CTAGCGGCGGCCGCGAGGTACAACGAGCCTGCCAAGAAASelV fw CTAGCGTTAATTAAGCAAGGGGTGGAGCTGGAGGAGTCSelV rev CTAGCGGCGGCCGCTGAGCTCACCCCAGCCCCAGCCACSelW fw CTAGCGTTAATTAACTTGACCCAGCCCCTCTCAGCAGASelW rev CTAGCGGCGGCCGCGCCCCCAACAGCAGCCACGAGAACASelX fw CTAGCGTTAATTAACCTGCCAGCCGCCCTGGCCCTSelX rev CTAGCGGCGGCCGCGCCTGCCTTCCCAGCAGGTCACAGASel15 fw CTAGCGTTAATTAAAGAGTGAAACATTCACAAAGATTTGCG Sel15 rev CTAGCGGCGGCCGCAAAACTCAAAAATTTGCCAAGTATGT SPS2 fw CTAGCGTTAATTAAACCTGCAACCATCTGACTTGGTCTCTGTSPS2 rev CTAGCGGCGGCCGCCACTGAGGCCTGGGCTCTGAGGCT
Supplementary Figure 1. List of primers used for PCR amplification of human SECIS elements and cloning downstream of luciferase UGA258 coding sequence.
SelX AUGA fw GGCCCTGGTCACTGCTCCGCTCTGGTCAAASelX AUGA rev TTTGACCAGAGCGGAGCAGTGACCAGGGCCSelX AAAC fw TCCGCTCTGGTCCCTTCCAGGCCAGCCAGAGSelX AAAC rev CTCTGGCTGGCCTGGAAGGGACCAGAGCGGAGpx3 AUGA fw AGGGGTGGCGTCTTCGGGAGGGGCCCAAAGGpx3 AUGA rev CTTTGGGCCCCTCCCGAAGACGCCACCCCTGpx3 AAAG fw GGGAGGGGCCCCCCTTGTGGGCGGACCTCCGpx3 AAAG rev GGAGGTCCGCCCACAAGGGGGGCCCCTCCCLucUAA258 fw GACTCGAAATCCTTATATCAAATATCLucUAA258 rev GATATTTGATATAAGGATTTCGAGTCLucUGU258 fw GACTCGAAATCCACATATCAAATATCLucUGU258 rev GATATTTGATATGTGGATTTCGAGTC
Supplementary Figure 2. List of primers used to generate mutations in luciferase UGA258-SelX and luciferase UGA258-Gpx3 constructs.
A
SelX Ch1Gpx3 Ch2 Ch3 Ch4 Ch5 Ch6 Ch7 Ch8 Ch9 Ch10 Ch11 Ch12
B
C
Ch1 CACGGACCCCATGGCAGGGGTGGCGTCTTCTGCATGATCCGCTCTGGTCAAACCCTTCCAGGCCAGCCAGAGTGGGGATGGTCTGAGGGGCCAGCCCTTAGTGCATCh2 CACGGACCCCATGGCAGGGGTGGCGTCTTCATGATCCGCTCTGGTCAAACCCTTCCAGGCCAGCCAGAGTGGGGAGCCTGTCTGAGGGGCCAGCCCTTAGTGCATCh3 CACGGACCCCATGGCAGGGGTGGCGTCTTCATGAGGGAGGGGCCCCAAACCCTTCCAGGCCAGGACCTCCCCTGAGCCTGTCTGAGGGGCCAGCCCTTAGTGCATCh4 CCTGCCAGCCGCCCTGGCCCTGGTCACTGCATGATCCGCTCTGGTAAAGCCCTTGTGGGCGCCAGAGTGGGGATGGTCTGTGACCTGCTGGGAAGGCAGGCCh5 CCTGCCAGCCGCCCTGGCCCTGGTCACTGCATGAGGGAGGGGCCCAAAGCCCTTGTGGGCGGACCTCCCCTGATGGTCTGTGACCTGCTGGGAAGGCAGGCCh6 CCTGCCAGCCGCCCTGGCCCTGGTCACATGAGGGAGGGGCCCAAAGCCCTTGTGGGCGGACCTCCCCTGAGCCTGTCTGTGACCTGCTGGGAAGGCAGGCCh7 CCTGCCAGCCGCCCTGGCCCTGGTCACATGATCCGCTCTGGTCAAACCCTTCCAGGCCAGCCAGAGTGGGGAGCCTGTCTGTGACCTGCTGGGAAGGCAGGCCh8 CACGGACCCCATGGCAGGGGTGGCGTCTTCTGCATGAGGGAGGGGCCCAAAGCCCTTGTGGGCGGACCTCCCCTGATGGTCTGAGGGGCCAGCCCTTAGTGCATCh9 CACGGACCCCATGGCAGGGGTGGCGTCTTCTGCATGATCCGCTCTGGTCAAACCCTTCCAGGCCAGCCAGAGTGGGGATGGTCTGAGGGGCCAGCCCTTAGTGCATCh10 CCTGCCAGCCGCCCTGGCCCTGGTCACATGAGGGAGGGGCCCAAAGCCCTTGTGGGCGGACCTCCCCTGAGCCTGTCTGTGACCTGCTGGGAAGGCAGGCCh11 CACGGACCCCATGGCAGGGGTGGCGTCTTCATGATCCGCTCTGGTAAAGCCCTTGTGGGCGCCAGAGTGGGGAGCCTGTCTGAGGGGCCAGCCCTTAGTGCATCh12 CCTGCCAGCCGCCCTGGCCCTGGTCACTGCATGAGGGAGGGGCCCCAAACCCTTCCAGGCCAGGACCTCCCCTGATGGTCTGTGACCTGCTGGGAAGGCAGGC
Ch1 fw: CACGGACCCCATGGCAGGGGTGGCGTCTTCTGCATGATCCGCTCTGGTCAAACCCTTCCAGGCCCh1 rev: ATGCACTAAGGGCTGGCCCCTCAGACCATCCCCACTCTGGCTGGCCTGGAAGGGTTTGACCAGACh2 fw: CACGGACCCCATGGCAGGGGTGGCGTCTTCATGATCCGCTCTGGTCAAACCCTTCCAGGCCAGCh2 rev: ATGCACTAAGGGCTGGCCCCTCAGACAGGCTCCCCACTCTGGCTGGCCTGGAAGGGTTTGACCACh3 fw: CACGGACCCCATGGCAGGGGTGGCGTCTTCATGAGGGAGGGGCCCCAAACCCTTCCAGGCCAGCh3 rev: ATGCACTAAGGGCTGGCCCCTCAGACAGGCTCAGGGGAGGTCCTGGCCTGGAAGGGTTTGGGGCCh4 fw: CCTGCCAGCCGCCCTGGCCCTGGTCACTGCATGATCCGCTCTGGTAAAGCCCTTGTGGGCGCh4 rev: GCCTGCCTTCCCAGCAGGTCACAGACCATCCCCACTCTGGCGCCCACAAGGGCTTTACCAGACh5 fw: CCTGCCAGCCGCCCTGGCCCTGGTCACTGCATGAGGGAGGGGCCCAAAGCCCTTGTGGGCGCh5 rev: GCCTGCCTTCCCAGCAGGTCACAGACCATCAGGGGAGGTCCGCCCACAAGGGCTTTGGGCCCCh6 fw: CCTGCCAGCCGCCCTGGCCCTGGTCACATGAGGGAGGGGCCCAAAGCCCTTGTGGGCGGACCh6 rev: GCCTGCCTTCCCAGCAGGTCACAGACAGGCTCAGGGGAGGTCCGCCCACAAGGGCTTTGGGCh7 fw: CCTGCCAGCCGCCCTGGCCCTGGTCACATGATCCGCTCTGGTCAAACCCTTCCAGGCCAGCCCh7 rev: GCCTGCCTTCCCAGCAGGTCACAGACAGGCTCCCCACTCTGGCTGGCCTGGAAGGGTTTGACCh8 fw: CACGGACCCCATGGCAGGGGTGGCGTCTTCTGCATGAGGGAGGGGCCCAAAGCCCTTGTGGGCCh8 rev: ATGCACTAAGGGCTGGCCCCTCAGACCATCAGGGGAGGTCCGCCCACAAGGGCTTTGGGCCCCCh9 fw: CACGGACCCCATGGCAGGGGTGGCGTCTTCTGCATGATCCGCTCTGGTCAAACCCTTCCAGGCCCh9 rev: ATGCACTAAGGGCTGGCCCCTCAGACCATCCCCACTCTGGCTGGCCTGGAAGGGTTTGACCAGACh10 fw: CCTGCCAGCCGCCCTGGCCCTGGTCACATGAGGGAGGGGCCCAAAGCCCTTGTGGGCGGACCh10 rev:GCCTGCCTTCCCAGCAGGTCACAGACAGGCTCAGGGGAGGTCCGCCCACAAGGGCTTTGGGCh11 fw: CACGGACCCCATGGCAGGGGTGGCGTCTTCATGATCCGCTCTGGTAAAGCCCTTGTGGGCGCCh11 rev:ATGCACTAAGGGCTGGCCCCTCAGACAGGCTCCCCACTCTGGCGCCCACAAGGGCTTTACCAGCh12 fw: CCTGCCAGCCGCCCTGGCCCTGGTCACTGCATGAGGGAGGGGCCCCAAACCCTTCCAGGCCACh12 rev:GCCTGCCTTCCCAGCAGGTCACAGACCATCAGGGGAGGTCCTGGCCTGGAAGGGTTTGGGGCC
Supplementary Figure 3. Structures, sequences and nomenclature of chimeric SECIS between SelX and Gpx3 cloned downstream of luciferase UGA258 coding sequence. (A) Schematic representation of the chimeric SECIS elements together with wild-type SelX and Gpx3. Structural domains from SelX and Gpx3 are represented in black and grey respectively. Ch: Chimera (B) Sequences of the chimeric SECIS elements. (C) List of overlapping primers used to generate chimerical SECIS elements
