Primer Sequences JB comments Designer Name of part Gene of origin Number Forward (5'-->3') Forward # BP Reverse (5'-->3') Reverse # BP Sequence of part without Added Restriction sites EXPECTED PCR Product of part with Added Restriction sites BAD Internal Restriction sites How the ORF will Be Read EXAMPLE PROMOTOR/ TERMINATOR JAMES Example promoter/terminator "GENE NAME [e.g. STE2, STE3, MFA1]" BBa_K11XXXX 5' GTTTCTTCGAATTCGCGGCCGCTTCTAGAG [20bp into the Sequence] 3' 50 5' GTTTCTTCCTGCAGCGGCCGCTACTAGTA [Reverse complement of 20 bp at end of given sequence] 3' 49 Sequence 5' GTT TCT TCG AAT TCG CGG CCG CTT CTA GAG [Sequence] TAC TAG TAG CGG CCG CTG CAG GAA GAA AC 3' NONE Left to Right EXAMPLE PROTEIN CODING SEQUENCE (PCS) JAMES Example PCS GENE NAME [e.g. vYFP] BBa_K11XXXX 5' GTTTCTTCGAATTCGCGGCCGCTTCTAG [ATG start and 20 bp into sequence]3' 50 5' GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTA [Reverse Complement of 20 bp into sequence WITHOUT STOP CODON] 3' 49 Sequence = ATG (start)... Sequence… STOP CODON REMOVED 5' GTTTCTTCGAATTCGCGGCCGCTTCTAG [ATG (start) then sequence with NO STOP] TAA TAATACTAGTAGCGGCCGCTGCAGGAAGAAAC3' NONE Left to Right OK DONE Jaime Two way Stop Between-bud 27-W FRS2-C + 200bp into each gene LtR BBa_K110001 gtttcttcgaattcgcggccgcttctagagGTAGCACTACATCCGGGCCGGCTACAGGA 59 gtttcttcctgcagcggccgctactagtaCCATGGGTCTACCAAAGGATCTAAGAGTC 58 GTAGCACTACATCCGGGCCGGCTACAGGAAGAGGTTGCTATAGAGTACAGACGGTTGAAAGAAGCTACCGCGTCGAAATGGCAATCATCATCACCCGCCGCACACACTGAAGGCGAATTGGAGCCTATTGATAAGTTCGGGAACCCGGTCAAGACAAGTCGTTTCCGCTCTCAACGGCTCCATATGGACAGTAAACCATAACCCATATGGAGACAGAAAGTAAATCAGAATCTATATTAACATATACGTATATATTTGTTTTGTGTACAATAACTACTACAAAATTGGGGACAGACGATATAAAACTAGAGGAACTGCTGAGCTTATTCGTACAAGTCTTCGTCCAATCTAGCAGCAGGATTGGTTTCGATAAAGTCCAAAGAGACTTTATGACCTAACAGTTCTCTGATGTTTTGAACCTTATATTTGATCATGGTAGGTCTTTCCAAGGATAACCCCCAACCAAGGACTCTTAGATCCTTTGGTAGACCCATGGACTCGAGCATTTCTGGTCTGAACATACC gtttcttcgaattcgcggccgcttctagagGTAGCACTACATCCGGGCCGGCTACAGGAAGAGGTTGCTATAGAGTACAGACGGTTGAAAGAAGCTACCGCGTCGAAATGGCAATCATCATCACCCGCCGCACACACTGAAGGCGAATTGGAGCCTATTGATAAGTTCGGGAACCCGGTCAAGACAAGTCGTTTCCGCTCTCAACGGCTCCATATGGACAGTAAACCATAACCCATATGGAGACAGAAAGTAAATCAGAATCTATATTAACATATACGTATATATTTGTTTTGTGTACAATAACTACTACAAAATTGGGGACAGACGATATAAAACTAGAGGAACTGCTGAGCTTATTCGTACAAGTCTTCGTCCAATCTAGCAGCAGGATTGGTTTCGATAAAGTCCAAAGAGACTTTATGACCTAACAGTTCTCTGATGTTTTGAACCTTATATTTGATCATGGTAGGTCTTTCCAAGGATAACCCCCAACCAAGGACTCTTAGATCCTTTGGTAGACCCATGGACTCGAGCATTTCTGGTCTGAACATACCtactagtagcggccgctgcaggaagaaac None Left to Right Smaller Version ---> DONE James Two way Stop Between-bud 27-W FRS2-C LtR BBa_K1100011 gtttcttcgaattcgcggccgcttctagagACCCATATGGAGACAGAAAGTAAATCAGA 59 gtttcttcctgcagcggccgctactagtaGCAGTTCCTCTAGTTTTATATCGTCTGTC 58 ACCCATATGGAGACAGAAAGTAAATCAGAATCTATATTAACATATACGTATATATTTGTTTTGTGTACAATAACTACTACAAAATTGGGGACAGACGATATAAAACTAGAGGAACTGCTGAGCT gtttcttcgaattcgcggccgcttctagagACCCATATGGAGACAGAAAGTAAATCAGAATCTATATTAACATATACGTATATATTTGTTTTGTGTACAATAACTACTACAAAATTGGGGACAGACGATATAAAACTAGAGGAACTGCTGAGCTtactagtagcggccgctgcaggaagaaac None Left to Right "OK, but bad sites; don't bother" DONE Jaime Two way Stop Between STE2-W and BST1-C +200bp into each gene LtR BBa_K110002 gtttcttcgaattcgcggccgcttctagagTCTGAGACTGCAGATGATAT 50 gtttcttcctgcagcggccgctactagtaATTTTTTTGGTTGGAAATAATTCTATT 56 TCTGAGACTGCAGATGATATAGAGAAAAATCAGTTTTATCAGTTGCCCACACCTACGAGTTCAAAAAATACTAGGATAGGACCGTTTGCTGATGCAAGTTACAAAGAGGGAGAAGTTGAACCCGTCGACATGTACACTCCCGATACGGCAGCTGATGAGGAAGCCAGAAAGTTCTGGACTGAAGATAATAATAATTTATGATCAAAATTTACGGCTTTGAAAAAGTAATTTCGTGACCTTCGGTATAAGGTTACTACTAGATTCAGGTGCTCATCAGATGCACCACATTCTCTATAAAAAAAAATGGTATCTTTCTTATTTGATAATATTTAAACTCCTTTACATAATAAACATCTCGTAAGTAGTGGTAGAAACCACCTTTGCTTTTACGAGTTCAAGCTTTTTTCTTGCCATGATCTAGAACTCTCAGGCAATATATACAGTTAATCTTTTTTTACTGGGTTGTAGTTCTAATGTATTGTTTCGAAAAATAGCAACCAGGCACATATTATGTTCACCAAGTGGTGTATCCAGTACAGATTTCTTATTCCATAAATGAAACTGAAAACGGTTAAGTACACAAATAAAATGATGGTAACTTTACCATCCCACGTGTCTAAAGGTACGCTGTTTGGCATTTTGAAAATAGAATTATTTCCAACCAAAAAAAT gtttcttcgaattcgcggccgcttctagagGTAGCACTACATCCGGGCCGGCTACAGGAAGAGGTTGCTATAGAGTACAGAC TCTGAGACTGCAGATGATATAGAGAAAAATCAGTTTTATCAGTTGCCCACACCTACGAGTTCAAAAAATACTAGGATAGGACCGTTTGCTGATGCAAGTTACAAAGAGGGAGAAGTTGAACCCGTCGACATGTACACTCCCGATACGGCAGCTGATGAGGAAGCCAGAAAGTTCTGGACTGAAGATAATAATAATTTATGATCAAAATTTACGGCTTTGAAAAAGTAATTTCGTGACCTTCGGTATAAGGTTACTACTAGATTCAGGTGCTCATCAGATGCACCACATTCTCTATAAAAAAAAATGGTATCTTTCTTATTTGATAATATTTAAACTCCTTTACATAATAAACATCTCGTAAGTAGTGGTAGAAACCACCTTTGCTTTTACGAGTTCAAGCTTTTTTCTTGCCATGATCTAGAACTCTCAGGCAATATATACAGTTAATCTTTTTTTACTGGGTTGTAGTTCTAATGTATTGTTTCGAAAAATAGCAACCAGGCACATATTATGTTCACCAAGTGGTGTATCCAGTACAGATTTCTTATTCCATAAATGAAACTGAAAACGGTTAAGTACACAAATAAAATGATGGTAACTTTACCATCCCACGTGTCTAAAGGTACGCTGTTTGGCATTTTGAAAATAGAATTATTTCCAACCAAAAAAATtactagtagcggccgctgcaggaagaaac Spe1 Xba1 Left to Right Smaller Version ---> DONE James Two way Stop Between STE2-W and BST1-C LtR BBa_K1100012 gtttcttcgaattcgcggccgcttctagagATCAAAATTTACGGCTTTGAAAAAGTAATT 60 gtttcttcctgcagcggccgctactagtaTCAAATAAGAAAGATACCATTTTTTTTTATAGAG 63 ATCAAAATTTACGGCTTTGAAAAAGTAATTTCGTGACCTTCGGTATAAGGTTACTACTAGATTCAGGTGCTCATCAGATGCACCACATTCTCTATAAAAAAAAATGGTATCTTTCTTATTTGA gtttcttcgaattcgcggccgcttctagagATCAAAATTTACGGCTTTGAAAAAGTAATTTCGTGACCTTCGGTATAAGGTTACTACTAGATTCAGGTGCTCATCAGATGCACCACATTCTCTATAAAAAAAAATGGTATCTTTCTTATTTGAtactagtagcggccgctgcaggaagaaac None Left to Right looks good DONE Jaime Two way Stop Between-SWP82-W and EMP47-C +200 into each gene LtR BBa_K110003 gtttcttcgaattcgcggccgcttctagagAACATTAAGGATTTTAAACCTCCATACATT 60 gtttcttcctgcagcggccgctactagtaAATCAGAAATTGGAGAAAGTCCTGGTTGAAC 60 AACATTAAGGATTTTAAACCTCCATACATTTATTCTCATGTGAACAAAGTACCACAAAATGTCGCTGGGGATAAAACGGCTGTTAAGCTGGACACTGAAGTAAAGAACACAAATGCTAATCCAGTGGTGGCGACGGATCCGGTCGCCGCTAAACCGGACAACCTGGCTAACTTCAGCAACGAAGTAGCTATGAATAATTGAAAGAAGTAAAACATAATGTAGGGAACACTCCAATAAGAAATAAACCATTGTTGAAGTAGTAATATAAACTATCGTTCGTTATTTTATAAAATATGTATTGTTACATATATATACATATATAGGGAATGCATGTGCACAATTGGTTTGAGTGGAAGGAATAATCCAAAATCATAGCAGTTTGGTCTTTATGATCTCTTGTCTGATTCTGAATGTGTAATATGCCATAACCAACATAATGAAAATTAATGGAAGTAACCAAATCATTAGTTTTCTGGCAATTTCGTCGACCTGAGGACCATCTTGTCCATGACGTTTGGTAGCTTCCCTATACTTTTCTTGTTCAACCAGGACTTTCTCCAATTTCTGATT gtttcttcgaattcgcggccgcttctagagAACATTAAGGATTTTAAACCTCCATACATTTATTCTCATGTGAACAAAGTACCACAAAATGTCGCTGGGGATAAAACGGCTGTTAAGCTGGACACTGAAGTAAAGAACACAAATGCTAATCCAGTGGTGGCGACGGATCCGGTCGCCGCTAAACCGGACAACCTGGCTAACTTCAGCAACGAAGTAGCTATGAATAATTGAAAGAAGTAAAACATAATGTAGGGAACACTCCAATAAGAAATAAACCATTGTTGAAGTAGTAATATAAACTATCGTTCGTTATTTTATAAAATATGTATTGTTACATATATATACATATATAGGGAATGCATGTGCACAATTGGTTTGAGTGGAAGGAATAATCCAAAATCATAGCAGTTTGGTCTTTATGATCTCTTGTCTGATTCTGAATGTGTAATATGCCATAACCAACATAATGAAAATTAATGGAAGTAACCAAATCATTAGTTTTCTGGCAATTTCGTCGACCTGAGGACCATCTTGTCCATGACGTTTGGTAGCTTCCCTATACTTTTCTTGTTCAACCAGGACTTTCTCCAATTTCTGATTtactagtagcggccgctgcaggaagaaac None Left to Right Smaller Version ---> DONE James Two way Stop Between-SWP82-W and EMP47-C LtR BBa_K1100013 gtttcttcgaattcgcggccgcttctagagAAAGAAGTAAAACATAATGTAGGGAACACTCC 62 gtttcttcctgcagcggccgctactagtaATTTTGGATTATTCCTTCCACTCAAACCAATTGTGC 65 AAAGAAGTAAAACATAATGTAGGGAACACTCCAATAAGAAATAAACCATTGTTGAAGTAGTAATATAAACTATCGTTCGTTATTTTATAAAATATGTATTGTTACATATATATACATATATAGGGAATGCATGTGCACAATTGGTTTGAGTGGAAGGAATAATCCAAAAT gtttcttcgaattcgcggccgcttctagagAAAGAAGTAAAACATAATGTAGGGAACACTCCAATAAGAAATAAACCATTGTTGAAGTAGTAATATAAACTATCGTTCGTTATTTTATAAAATATGTATTGTTACATATATATACATATATAGGGAATGCATGTGCACAATTGGTTTGAGTGGAAGGAATAATCCAAAATtactagtagcggccgctgcaggaagaaac None Left to Right "Bad sites, don't bother" DONE Ambhi Alpha- Promoter Ste3 LtR BBa_K110004 gtttcttcgaattcgcggccgcttctagagCCAAAAAAATGCAATTATCAGA 52 gtttcttcctgcagcggccgctactagtaGAAAATTTTGATAGTATTTTGC 51 CCAAAAAAATGCAATTATCAGAACAAAACTATTTAAGGTGACTTCTCCGTATTTACACCACCAGAAGCGTTCTGGCTCCCCTTTTCTCTAAACGTTAAACATTTTACAATTGAAATGTTACCAATCCTATATTATTGTACCACATTGCCAGATTTATGAACTCTGGGTATGGGTGCTAATTTTCGTTAGAAGCGCTGGTACAATTTTCTCTGTCATTGTGACACTAATTAGGAAACTTCTCGACTATCAATGTGTAAATGAAGGAATAATGGCGGAAACTTTGAAACTTTGTCAATAATTGCATCATTGGATGCGTTTCATTTGGCCGTTATCACGGAGAGGCAGAGTTCTCTCCACAATTTGGGCAGAAGTCTTTTGAAAAGACATATATATATATATATATGTATATGAGTGGATGCTTAAGGTAAGAATAATTTCTGAATTCCCAAGTATTCATTTTGTGCAGTATTCACATATTCTATTTTATTGCTTTTTAACTTTAGAGGCAATTAAATTTGTGTAGGAAAGGCAAAATACTATCAAAATTTTC gtttcttcgaattcgcggccgcttctagagCCAAAAAAATGCAATTATCAGAACAAAACTATTTAAGGTGACTTCTCCGTATTTACACCACCAGAAGCGTTCTGGCTCCCCTTTTCTCTAAACGTTAAACATTTTACAATTGAAATGTTACCAATCCTATATTATTGTACCACATTGCCAGATTTATGAACTCTGGGTATGGGTGCTAATTTTCGTTAGAAGCGCTGGTACAATTTTCTCTGTCATTGTGACACTAATTAGGAAACTTCTCGACTATCAATGTGTAAATGAAGGAATAATGGCGGAAACTTTGAAACTTTGTCAATAATTGCATCATTGGATGCGTTTCATTTGGCCGTTATCACGGAGAGGCAGAGTTCTCTCCACAATTTGGGCAGAAGTCTTTTGAAAAGACATATATATATATATATATGTATATGAGTGGATGCTTAAGGTAAGAATAATTTCTGAATTCCCAAGTATTCATTTTGTGCAGTATTCACATATTCTATTTTATTGCTTTTTAACTTTAGAGGCAATTAAATTTGTGTAGGAAAGGCAAAATACTATCAAAATTTTCtactagtagcggccgctgcaggaagaaac EcoR1 Left to Right OK DONE Ambhi Alpha- Promoter MFalpha2 LtR BBa_K110005 gtttcttcgaattcgcggccgcttctagagTGCTCATTGATGTCCCTAAAAGGTAATGT 59 gtttcttcctgcagcggccgctactagtaTTTCTCCAATATGTGAATTTACTGGAAT 57 TGCTCATTGATGTCCCTAAAAGGTAATGTCGCATAGTAAGATGTATGTCCCATATCAAACGCATTCCTAGAAAAACATGCACTGAATGAATAAGCAGTTACTTCGATTACTAATCTTCACAGCATAATTTTTCTCTTATCTTGTAAACCTCTTTCTATCTTTTCATAAAGTTTGTCACTTATATTTCTTCATTGGTACATCAATGCCAGCAACGATGTGCGCATCTGGGCGACGCCTGTAGTGATTGTTTTCAAGGTATCGAGCCAAACTATTCATCGTTACTGTTTCAAATATTCAGTTGTTTCAGTACAGAGTCGCCGTGGACCTAGTGAAACTTGGTGTCTTTACAGCGCAGAGACGAGGGCTTATATGTATAAAAGCTGTCCTTGATTCTGGTGTAGTTTGAGGTGTCCTTCCTATATCTGTTTTTATATTCTATATAATGGATAATTACTACCATCACCTGCATCAAATTCCAGTAAATTCACATATTGGAGAAA gtttcttcgaattcgcggccgcttctagagTGCTCATTGATGTCCCTAAAAGGTAATGTCGCATAGTAAGATGTATGTCCCATATCAAACGCATTCCTAGAAAAACATGCACTGAATGAATAAGCAGTTACTTCGATTACTAATCTTCACAGCATAATTTTTCTCTTATCTTGTAAACCTCTTTCTATCTTTTCATAAAGTTTGTCACTTATATTTCTTCATTGGTACATCAATGCCAGCAACGATGTGCGCATCTGGGCGACGCCTGTAGTGATTGTTTTCAAGGTATCGAGCCAAACTATTCATCGTTACTGTTTCAAATATTCAGTTGTTTCAGTACAGAGTCGCCGTGGACCTAGTGAAACTTGGTGTCTTTACAGCGCAGAGACGAGGGCTTATATGTATAAAAGCTGTCCTTGATTCTGGTGTAGTTTGAGGTGTCCTTCCTATATCTGTTTTTATATTCTATATAATGGATAATTACTACCATCACCTGCATCAAATTCCAGTAAATTCACATATTGGAGAAAtactagtagcggccgctgcaggaagaaac None Left to Right looks good! DONE Ambhi Alpha- Promoter MFalpha1 LtR BBa_K110006 gtttcttcgaattcgcggccgcttctagagTAGTGTGGATTTCAATAATTTCCG 54 gtttcttcctgcagcggccgctactagtaTTCTTTTAATCGTTTATATTGTGTATGAAA 59 TAGTGTGGATTTCAATAATTTCCGAATTAGGAATAAATGCGCTAAATAGACATCCCGTTCTCTTTGGTAATCTGCATAATTCTGATGCAATATCCAACAACTATTTGTGCAATTATTTAACAAAATCCAATTAACTTTCCTAATTAGTCCTTCAATAGAACATCTGTATTCCTTTTTTTTATGAACACCTTCCTAATTAGGCCATCAACGACAGTAAATTTTGCCGAATTTAATAGCTTCTACTGAAAAACAGTGGACCATGTGAAAAGATGCATCTCATTTATCAAACACATAATATTCAAGTGAGCCTTACTTCAATTGTATTGAAGTGCAAGAAAACCAAAAAGCAACAACAGGTTTTGGATAAGTACATATATAAGAGGGCCTTTTGTTCCCATCAAAAATGTTACTGTTCTTACGATTCATTTACGATTCAAGAATAGTTCAAACAAGAAGATTACAAACTATCAATTTCATACACAATATAAACGATTAAAAGAA gtttcttcgaattcgcggccgcttctagagTAGTGTGGATTTCAATAATTTCCGAATTAGGAATAAATGCGCTAAATAGACATCCCGTTCTCTTTGGTAATCTGCATAATTCTGATGCAATATCCAACAACTATTTGTGCAATTATTTAACAAAATCCAATTAACTTTCCTAATTAGTCCTTCAATAGAACATCTGTATTCCTTTTTTTTATGAACACCTTCCTAATTAGGCCATCAACGACAGTAAATTTTGCCGAATTTAATAGCTTCTACTGAAAAACAGTGGACCATGTGAAAAGATGCATCTCATTTATCAAACACATAATATTCAAGTGAGCCTTACTTCAATTGTATTGAAGTGCAAGAAAACCAAAAAGCAACAACAGGTTTTGGATAAGTACATATATAAGAGGGCCTTTTGTTCCCATCAAAAATGTTACTGTTCTTACGATTCATTTACGATTCAAGAATAGTTCAAACAAGAAGATTACAAACTATCAATTTCATACACAATATAAACGATTAAAAGAAtactagtagcggccgctgcaggaagaaac None Left to Right OK DONE Rick A Promoter MFA2 LtR BBa_K110007 gtttcttcgaattcgcggccgcttctagagAAAGAATTCAAACTCAGTTAGTAATAATAA 60 gtttcttcctgcagcggccgctactagtaTAAGGTTGGTAGTTATTGTTGTATGAAGATG 60 AAAGAATTCAAACTCAGTTAGTAATAATAACAGGCCCTCATCTATTTTCTCGGAAAACTTGGTGGATTTTGATGATGTTAACATGGTCGATAAGACCAGACTGTTTATTTTTTTATTTTTCAGTTTCATCATTACTATACCGTTTATGGTATAAATTAGAAAAGTTAAAGCAGCATGTTTTCATTTGAAACAAATACTAATGCAATAATAGGTACACCATCTACTACATAATTAATTGATAGTTTCCTTTTCCGTTAAGTGCATGCATAGGACGCCCATAATTTTTTAAGTTAAAAAAGCATGTATTTACCTATTCGGGAAATTTACATGACATGGATGCCATAAGGAACGAAAATGAAACATGCATGTCAGAGGAAAAAGAACAAAGCGAGAGGAAAAAGCTGTTGCATTACCACGTAATTTTGTATATAAATATCTGATAAATAACCATTTTATTTCCATCCACTTCTTCTGTCGTTCATCCGTTCATTGACATCACTAGAGACACCAGCGAGCTATCATCTTCATACAACAATAACTACCAACCTTA gtttcttcgaattcgcggccgcttctagagAAAGAATTCAAACTCAGTTAGTAATAATAACAGGCCCTCATCTATTTTCTCGGAAAACTTGGTGGATTTTGATGATGTTAACATGGTCGATAAGACCAGACTGTTTATTTTTTTATTTTTCAGTTTCATCATTACTATACCGTTTATGGTATAAATTAGAAAAGTTAAAGCAGCATGTTTTCATTTGAAACAAATACTAATGCAATAATAGGTACACCATCTACTACATAATTAATTGATAGTTTCCTTTTCCGTTAAGTGCATGCATAGGACGCCCATAATTTTTTAAGTTAAAAAAGCATGTATTTACCTATTCGGGAAATTTACATGACATGGATGCCATAAGGAACGAAAATGAAACATGCATGTCAGAGGAAAAAGAACAAAGCGAGAGGAAAAAGCTGTTGCATTACCACGTAATTTTGTATATAAATATCTGATAAATAACCATTTTATTTCCATCCACTTCTTCTGTCGTTCATCCGTTCATTGACATCACTAGAGACACCAGCGAGCTATCATCTTCATACAACAATAACTACCAACCTTAtactagtagcggccgctgcaggaagaaac None Left to Right OK except termini are reversed on column L DONE Rick A Promoter MFA2 RtL BBa_K1100014 gtttcttcgaattcgcggccgcttctagagTAAGGTTGGTAGTTATTGTTGTATGAAGAT 60 gtttcttcctgcagcggccgctactagtaAAAGAATTCAAACTCAGTTAGTAATAATAAC 60 AAAGAATTCAAACTCAGTTAGTAATAATAACAGGCCCTCATCTATTTTCTCGGAAAACTTGGTGGATTTTGATGATGTTAACATGGTCGATAAGACCAGACTGTTTATTTTTTTATTTTTCAGTTTCATCATTACTATACCGTTTATGGTATAAATTAGAAAAGTTAAAGCAGCATGTTTTCATTTGAAACAAATACTAATGCAATAATAGGTACACCATCTACTACATAATTAATTGATAGTTTCCTTTTCCGTTAAGTGCATGCATAGGACGCCCATAATTTTTTAAGTTAAAAAAGCATGTATTTACCTATTCGGGAAATTTACATGACATGGATGCCATAAGGAACGAAAATGAAACATGCATGTCAGAGGAAAAAGAACAAAGCGAGAGGAAAAAGCTGTTGCATTACCACGTAATTTTGTATATAAATATCTGATAAATAACCATTTTATTTCCATCCACTTCTTCTGTCGTTCATCCGTTCATTGACATCACTAGAGACACCAGCGAGCTATCATCTTCATACAACAATAACTACCAACCTTA gtttcttcgaattcgcggccgcttctagagTAAGGTTGGTAGTTATTGTTGTATGAAGATGATAGCTCGCTGGTGTCTCTAGTGATGTCAATGAACGGATGAACGACAGAAGAAGTGGATGGAAATAAAATGGTTATTTATCAGATATTTATATACAAAATTACGTGGTAATGCAACAGCTTTTTCCTCTCGCTTTGTTCTTTTTCCTCTGACATGCATGTTTCATTTTCGTTCCTTATGGCATCCATGTCATGTAAATTTCCCGAATAGGTAAATACATGCTTTTTTAACTTAAAAAATTATGGGCGTCCTATGCATGCACTTAACGGAAAAGGAAACTATCAATTAATTATGTAGTAGATGGTGTACCTATTATTGCATTAGTATTTGTTTCAAATGAAAACATGCTGCTTTAACTTTTCTAATTTATACCATAAACGGTATAGTAATGATGAAACTGAAAAATAAAAAAATAAACAGTCTGGTCTTATCGACCATGTTAACATCATCAAAATCCACCAAGTTTTCCGAGAAAATAGATGAGGGCCTGTTATTATTACTAACTGAGTTTGAATTCTTTtactagtagcggccgctgcaggaagaaac None Right to Left promoter segment stops at wrong ATG. Sequence needs to be shifted 60 bp to the right; MFA1 protein starts MQPSTATAAPKEKTSSEKKDNYIIKGVFWDPACVIA DONE Rick A Promoter MFA1 LtR BBa_K110008 gtttcttcgaattcgcggccgcttctagagGAAAGGTTCAGTATACAATGACCCATTCAT 60 gtttcttcctgcagcggccgctactagtaTAAGGTTGGTAGTTATTGTTGTATGAAGATG 60 AAGAATTTATTGCGTCTGGGTTCAACACTAATTATGCGTACGAAAGGGTGTTGACAGAGGCATTTATGGGCTTAGGATGTGTTATATCCGAGGAGCTTTAAAACATCAGGATAGTGTGCAACGTGGCATAAGCTATGTAATCAACTACTTTTTATTTTCTATGTACGCATATACATGCATTCACGATCTGTTTCAGTGTTCAGAAAAAAGGCACCTACTGCTACGGTTGGCCCATACCTTTATTCTTTGTTCTTGTTACAAACGAGTGTGTAATTACCCAAAAAGGAAATTTACATGTTAAATGAAACCCAGTAATCAGAAAAAACAGTTAAGAAACCTAAAATGGTAGAGATAAAGATACAGATTCAGTGGTTGCTGAAAATCAAGTAAAAAAATGAAATAGAGTCTTCATATATAAACCGCCAGAAATGAATTA gtttcttcgaattcgcggccgcttctagagAAAGAATTCAAACTCAGTTAGTAATAATAACAGGCCCTCATCTATTTTCTCGGAAAACTTGGTGGATTTTGATGATGTTAACATGGTCGATAAGACCAGACTGTTTATTTTTTTATTTTTCAGTTTCATCATTACTATACCGTTTATGGTATAAATTAGAAAAGTTAAAGCAGCATGTTTTCATTTGAAACAAATACTAATGCAATAATAGGTACACCATCTACTACATAATTAATTGATAGTTTCCTTTTCCGTTAAGTGCATGCATAGGACGCCCATAATTTTTTAAGTTAAAAAAGCATGTATTTACCTATTCGGGAAATTTACATGACATGGATGCCATAAGGAACGAAAATGAAACATGCATGTCAGAGGAAAAAGAACAAAGCGAGAGGAAAAAGCTGTTGCATTACCACGTAATTTTGTATATAAATATCTGATAAATAACCATTTTATTTCCATCCACTTCTTCTGTCGTTCATCCGTTCATTGACATCACTAGAGACACCAGCGAGCTATCATCTTCATACAACAATAACTACCAACCTTAtactagtagcggccgctgcaggaagaaac None Left to Right see above; also termini reversed on L DONE Rick A Promoter MFA1 RtL BBa_K1100015 gtttcttcgaattcgcggccgcttctagagTTCTATTCGATGGCTTTGTACTTATTTTGG 60 gtttcttcctgcagcggccgctactagtaGAAAGGTTCAGTATACAATGACCCATTCATC 60 AAGAATTTATTGCGTCTGGGTTCAACACTAATTATGCGTACGAAAGGGTGTTGACAGAGGCATTTATGGGCTTAGGATGTGTTATATCCGAGGAGCTTTAAAACATCAGGATAGTGTGCAACGTGGCATAAGCTATGTAATCAACTACTTTTTATTTTCTATGTACGCATATACATGCATTCACGATCTGTTTCAGTGTTCAGAAAAAAGGCACCTACTGCTACGGTTGGCCCATACCTTTATTCTTTGTTCTTGTTACAAACGAGTGTGTAATTACCCAAAAAGGAAATTTACATGTTAAATGAAACCCAGTAATCAGAAAAAACAGTTAAGAAACCTAAAATGGTAGAGATAAAGATACAGATTCAGTGGTTGCTGAAAATCAAGTAAAAAAATGAAATAGAGTCTTCATATATAAACCGCCAGAAATGAATTA gtttcttcgaattcgcggccgcttctagagTTCTATTCGATGGCTTTGTACTTATTTTGGTTTTATCCGAGAAACAGTTACAGATCCCTCTCATTAATTCATTTCTGGCGGTTTATATATGAAGACTCTATTTCATTTTTTTACTTGATTTTCAGCAACCACTGAATCTGTATCTTTATCTCTACCATTTTAGGTTTCTTAACTGTTTTTTCTGATTACTGGGTTTCATTTAACATGTAAATTTCCTTTTTGGGTAATTACACACTCGTTTGTAACAAGAACAAAGAATAAAGGTATGGGCCAACCGTAGCAGTAGGTGCCTTTTTTCTGAACACTGAAACAGATCGTGAATGCATGTATATGCGTACATAGAAAATAAAAAGTAGTTGATTACATAGCTTATGCCACGTTGCACACTATCCTGATGTTTTAAAGCTCCTCGGATATAACACATCCTAAGCCCATAAATGCCTCTGTCAACACCCTTTCGTACGCATAATTAGTGTTGAACCCAGACGCAATAAATTCTTTCCTGTGCTCAAGATCTTTGATGAATGGGTCATTGTATACTGAACCTTTCtactagtagcggccgctgcaggaagaaac None Right to Left OK DONE Rick A Promoter Ste2 LtR BBa_K110009 gtttcttcgaattcgcggccgcttctagagAGAACGAACTGTAGAATAGTCCGGATATG 59 gtttcttcctgcagcggccgctactagtaTTTTTGATTCTTGGATATGGTTCTTAAC 57 AGAACGAACTGTAGAATAGTCCGGATATGTTATCCAATGCCTGCCAAAATGCATTGTCACACGCTGTAGTGCTCGAATAGGTGTTGCAATCCGTCAATATACGTCTTGCTCTGTGGGTAAATGTCTCGTGCATTAAGACAGGCTAGTATAAACGAGAAGAAGTATCCTGCTTTGCAATGAAACAATAGTATCCGCTAAGAATTTAAGCAGGCCAACGTCCATACTGCTTAGGACCTGTGCCTGGCAAGTCGCAGATTGAAGTTTTTTCAACCATGTAAATTTCCTAATTGGGTAAGTACATGATGAAACACATATGAAGAAAAAAGCTTTCCTACATATTCAAGATTTTTTTCTGTGGGTGGAATACTATTTAAGGAGTGCTATTAGTATCTTATTTGACTTCAAAGCAATACGATACCTTTTCTTTTCACCTGCTCTGGCTATAATTATAATTGGTTACTTAAAAATGCACCGTTAAGAACCATATCCAAGAATCAAAA gtttcttcctgcagcggccgctactagtaAGAACGAACTGTAGAATAGTCCGGATATGTTATCCAATGCCTGCCAAAATGCATTGTCACACGCTGTAGTGCTCGAATAGGTGTTGCAATCCGTCAATATACGTCTTGCTCTGTGGGTAAATGTCTCGTGCATTAAGACAGGCTAGTATAAACGAGAAGAAGTATCCTGCTTTGCAATGAAACAATAGTATCCGCTAAGAATTTAAGCAGGCCAACGTCCATACTGCTTAGGACCTGTGCCTGGCAAGTCGCAGATTGAAGTTTTTTCAACCATGTAAATTTCCTAATTGGGTAAGTACATGATGAAACACATATGAAGAAAAAAGCTTTCCTACATATTCAAGATTTTTTTCTGTGGGTGGAATACTATTTAAGGAGTGCTATTAGTATCTTATTTGACTTCAAAGCAATACGATACCTTTTCTTTTCACCTGCTCTGGCTATAATTATAATTGGTTACTTAAAAATGCACCGTTAAGAACCATATCCAAGAATCAAAAAtactagtagcggccgctgcaggaagaaac None Left to Right Termini reversed DONE Rick A Promoter Ste2 RtL BBa_K1100016 gtttcttcgaattcgcggccgcttctagagTTTTTGATTCTTGGATATGGTTCTTAACGGT 61 gtttcttcctgcagcggccgctactagtaAGAACGAACTGTAGAATAGTCCGGATATGTT 60 AGAACGAACTGTAGAATAGTCCGGATATGTTATCCAATGCCTGCCAAAATGCATTGTCACACGCTGTAGTGCTCGAATAGGTGTTGCAATCCGTCAATATACGTCTTGCTCTGTGGGTAAATGTCTCGTGCATTAAGACAGGCTAGTATAAACGAGAAGAAGTATCCTGCTTTGCAATGAAACAATAGTATCCGCTAAGAATTTAAGCAGGCCAACGTCCATACTGCTTAGGACCTGTGCCTGGCAAGTCGCAGATTGAAGTTTTTTCAACCATGTAAATTTCCTAATTGGGTAAGTACATGATGAAACACATATGAAGAAAAAAGCTTTCCTACATATTCAAGATTTTTTTCTGTGGGTGGAATACTATTTAAGGAGTGCTATTAGTATCTTATTTGACTTCAAAGCAATACGATACCTTTTCTTTTCACCTGCTCTGGCTATAATTATAATTGGTTACTTAAAAATGCACCGTTAAGAACCATATCCAAGAATCAAAA gtttcttcctgcagcggccgctactagtaTTTTTGATTCTTGGATATGGTTCTTAACGGTGCATTTTTAAGTAACCAATTATAATTATAGCCAGAGCAGGTGAAAAGAAAAGGTATCGTATTGCTTTGAAGTCAAATAAGATACTAATAGCACTCCTTAAATAGTATTCCACCCACAGAAAAAAATCTTGAATATGTAGGAAAGCTTTTTTCTTCATATGTGTTTCATCATGTACTTACCCAATTAGGAAATTTACATGGTTGAAAAAACTTCAATCTGCGACTTGCCAGGCACAGGTCCTAAGCAGTATGGACGTTGGCCTGCTTAAATTCTTAGCGGATACTATTGTTTCATTGCAAAGCAGGATACTTCTTCTCGTTTATACTAGCCTGTCTTAATGCACGAGACATTTACCCACAGAGCAAGACGTATATTGACGGATTGCAACACCTATTCGAGCACTACAGCGTGTGACAATGCATTTTGGCAGGCATTGGATAACATATCCGGACTATTCTACAGTTCGTTCTtactagtagcggccgctgcaggaagaaac None Right to Left "reference sequence not included in ""package""; I have no way to confirm this iis really sapphire as opposed to something else" The Restriction Sites were switched in this protein as it should be ligated into the plasmid in the reverse direction- see plasmid drawing Pavel/James Sapphire Fluorescent Protein yESapphire LtR BBa_K110010 gtttcttcgaattcgcggccgcttctagagATGTCTAAAGGTGAAGAATTATTCACTGGTGTTG 64 gtttcttcctgcagcggccgctactagtattaTTATTTGTACAATTCATCCATACCATGGG 61 ATGTCTAAAGGTGAAGAATTATTCACTGGTGTTGTCCCAATTTTGGTTGAATTAGATGGTGATGTTAATGGTCACAAATTTTCTGTCTCCGGTGAAGGTGAAGGTGATGCTACTTACGGTAAATTGACCTTAAAATTTATTTGTACTACTGGTAAATTGCCAGTTCCATGGCCAACCTTAGTCACTACTTTTTCTTATGGTGTTCAATGTTTTGCTAGATACCCAGATCATATGAAACAACATGACTTTTTCAAGTCTGCCATGCCAGAAGGTTATGTTCAAGAAAGAACTATTTTTTTCAAAGATGACGGTAACTACAAGACCAGAGCTGAAGTCAAGTTTGAAGGTGATACCTTAGTTAATAGAATCGAATTAAAAGGTATTGATTTTAAAGAAGATGGTAACATTTTAGGTCACAAATTGGAATACAACTTTAACTCTCACAATGTTTACATCATGGCTGACAAACAAAAGAATGGTATCAAAGTTAACTTCAAAATTAGACACAACATTGAAGATGGTTCTGTTCAATTAGCTGACCATTATCAACAAAATACTCCAATTGGTGATGGTCCAGTCTTGTTACCAGACAACCATTACTTATCCATTCAATCTGCCTTATCCAAAGATCCAAACGAAAAGAGAGACCACATGGTCTTGTTAGAATTTGTTACTGCTGCTGGTATTACCCATGGTATGGATGAATTGTACAAATAA gtttcttcgaattcgcggccgcttctagagATGTCTAAAGGTGAAGAATTATTCACTGGTGTTGTCCCAATTTTGGTTGAATTAGATGGTGATGTTAATGGTCACAAATTTTCTGTCTCCGGTGAAGGTGAAGGTGATGCTACTTACGGTAAATTGACCTTAAAATTTATTTGTACTACTGGTAAATTGCCAGTTCCATGGCCAACCTTAGTCACTACTTTTTCTTATGGTGTTCAATGTTTTGCTAGATACCCAGATCATATGAAACAACATGACTTTTTCAAGTCTGCCATGCCAGAAGGTTATGTTCAAGAAAGAACTATTTTTTTCAAAGATGACGGTAACTACAAGACCAGAGCTGAAGTCAAGTTTGAAGGTGATACCTTAGTTAATAGAATCGAATTAAAAGGTATTGATTTTAAAGAAGATGGTAACATTTTAGGTCACAAATTGGAATACAACTTTAACTCTCACAATGTTTACATCATGGCTGACAAACAAAAGAATGGTATCAAAGTTAACTTCAAAATTAGACACAACATTGAAGATGGTTCTGTTCAATTAGCTGACCATTATCAACAAAATACTCCAATTGGTGATGGTCCAGTCTTGTTACCAGACAACCATTACTTATCCATTCAATCTGCCTTATCCAAAGATCCAAACGAAAAGAGAGACCACATGGTCTTGTTAGAATTTGTTACTGCTGCTGGTATTACCCATGGTATGGATGAATTGTACAAATAAtactagtagcggccgctgcaggaagaaac None Left to Right Termini reversed Pavel/James Sapphire Fluorescent Protein yESapphire RtL BBa_K1100017 gtttcttcgaattcgcggccgcttctagagttaTTATTTGTACAATTCATCCATACCATGGGT 63 gtttcttcctgcagcggccgctactagtaATGTCTAAAGGTGAAGAATTATTCACTGGT 59 ATGTCTAAAGGTGAAGAATTATTCACTGGTGTTGTCCCAATTTTGGTTGAATTAGATGGTGATGTTAATGGTCACAAATTTTCTGTCTCCGGTGAAGGTGAAGGTGATGCTACTTACGGTAAATTGACCTTAAAATTTATTTGTACTACTGGTAAATTGCCAGTTCCATGGCCAACCTTAGTCACTACTTTTTCTTATGGTGTTCAATGTTTTGCTAGATACCCAGATCATATGAAACAACATGACTTTTTCAAGTCTGCCATGCCAGAAGGTTATGTTCAAGAAAGAACTATTTTTTTCAAAGATGACGGTAACTACAAGACCAGAGCTGAAGTCAAGTTTGAAGGTGATACCTTAGTTAATAGAATCGAATTAAAAGGTATTGATTTTAAAGAAGATGGTAACATTTTAGGTCACAAATTGGAATACAACTTTAACTCTCACAATGTTTACATCATGGCTGACAAACAAAAGAATGGTATCAAAGTTAACTTCAAAATTAGACACAACATTGAAGATGGTTCTGTTCAATTAGCTGACCATTATCAACAAAATACTCCAATTGGTGATGGTCCAGTCTTGTTACCAGACAACCATTACTTATCCATTCAATCTGCCTTATCCAAAGATCCAAACGAAAAGAGAGACCACATGGTCTTGTTAGAATTTGTTACTGCTGCTGGTATTACCCATGGTATGGATGAATTGTACAAATAA gtttcttcgaattcgcggccgcttctagagttaTTATTTGTACAATTCATCCATACCATGGGTAATACCAGCAGCAGTAACAAATTCTAACAAGACCATGTGGTCTCTCTTTTCGTTTGGATCTTTGGATAAGGCAGATTGAATGGATAAGTAATGGTTGTCTGGTAACAAGACTGGACCATCACCAATTGGAGTATTTTGTTGATAATGGTCAGCTAATTGAACAGAACCATCTTCAATGTTGTGTCTAATTTTGAAGTTAACTTTGATACCATTCTTTTGTTTGTCAGCCATGATGTAAACATTGTGAGAGTTAAAGTTGTATTCCAATTTGTGACCTAAAATGTTACCATCTTCTTTAAAATCAATACCTTTTAATTCGATTCTATTAACTAAGGTATCACCTTCAAACTTGACTTCAGCTCTGGTCTTGTAGTTACCGTCATCTTTGAAAAAAATAGTTCTTTCTTGAACATAACCTTCTGGCATGGCAGACTTGAAAAAGTCATGTTGTTTCATATGATCTGGGTATCTAGCAAAACATTGAACACCATAAGAAAAAGTAGTGACTAAGGTTGGCCATGGAACTGGCAATTTACCAGTAGTACAAATAAATTTTAAGGTCAATTTACCGTAAGTAGCATCACCTTCACCTTCACCGGAGACAGAAAATTTGTGACCATTAACATCACCATCTAATTCAACCAAAATTGGGACAACACCAGTGAATAATTCTTCACCTTTAGACATtactagtagcggccgctgcaggaagaaac None Right to Left no primers needed; existing part iGEM Venus Yellow Fluorescent Protein EYFP BBa_J63001 gtttcttcgaattcgcggccgcttctagagGCAACTAGCGGCATGGTTAGTA 52 gtttcttcctgcagcggccgctactagtaTGCGGTACCAGAACCTTTGTAT 51 "GCAACTAGCGGCATGGTTAGTAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTTTAGTTGAACTAGATGGCGACGTGAACGGTCATAAGTTCAGTGTCTCCGGCGAAGGTGAGGGTGATGCAACGTACGGTAAGTTAACTTTGAAGTTAATATGTACAACCGGCAAGCTGCCTGTTCCCTGGCCTACCCTGGTGACAACGTTAGGTTATGGGTTGATGTGCTTTGCTAGATACCCAGATCACATGAAAAGGCATGACTTCTTTAAATCTGCAATGCCAGAAGGTTACGTCCAAGAACGTACTATTTTCTTTAAAGATGACGGTAATTATAAAACTAGGGCTGAAGTTAAATTCGAAGGTGACACACTTGTAAATCGAATAGAGTTAAAGGGGATTGATTTCAAAGAGGATGGTAATATTCTAGGCCATAAACTTGAATATAACTAT AATTCACACAACGTTTACATTACCGCCGACAAGCAGAAGAATGGAATCAAAGCCAATTTTAAGATTAGACACAATATTGAGGATGGTGGAGTACAGCTTGCTGATCATTACCAACAAAATACCCCGATCGGTGATGGACCAGTTTTGCTACCCGATAACCATTATCTGTCCTATCAAAGCAAATTGTCAAAAGATCCTAACGAAAAAAGAGACCACATGGTACTCTTGGAATTTGTAACAGCTGCTGGGATTACACATGGCATGGATGAACTATACAAAGGTTCTGGTACCGCA " "gtttcttcgaattcgcggccgcttctagagGCAACTAGCGGCATGGTTAGTAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTTTAGTTGAACTAGATGGCGACGTGAACGGTCATAAGTTCAGTGTCTCCGGCGAAGGTGAGGGTGATGCAACGTACGGTAAGTTAACTTTGAAGTTAATATGTACAACCGGCAAGCTGCCTGTTCCCTGGCCTACCCTGGTGACAACGTTAGGTTATGGGTTGATGTGCTTTGCTAGATACCCAGATCACATGAAAAGGCATGACTTCTTTAAATCTGCAATGCCAGAAGGTTACGTCCAAGAACGTACTATTTTCTTTAAAGATGACGGTAATTATAAAACTAGGGCTGAAGTTAAATTCGAAGGTGACACACTTGTAAATCGAATAGAGTTAAAGGGGATTGATTTCAAAGAGGATGGTAATATTCTAGGCCATAAACTTGAATATAACTATAATTCACACAACGTTTACATTACCGCCGACAAGCAGAAGAATGGAATCAAAGCCAATTTTAAGATTAGACACAATATTGAGGATGGTGGAGTACAGCTTGCTGATCATTACCAACAAAATACCCCGATCGGTGATGGACCAGTTTTGCTACCCGATAACCATTATCTGTCCTATCAAAGCAAATTGTCAAAAGATCCTAACGAAAAAAGAGACCACATGGTACTCTTGGAATTTGTAACAGCTGCTGGGATTACACATGGCATGGATGAACTATACAAAGGTTCTGGTACCGCAtactagtagcggccgctgcaggaagaaac " None Left to Right