Essential Basic Part Types Coding Sequences C Complete

Essential Basic Part Types Coding Sequences (C) - Complete open reading frames (type I), or sequences encoding polypeptides but lacking either a stop codon (type II), a start codon (type III), or both (type IV) Ribosome Binding Sites (RBS) - Sequence encoding a ribosome binding site, fused 5' to an ORF part Terminators (TT) - Sequence causing transcription termination (and more can come later, with their own part definitions and standards rules)

Potter Standard Polylinker GAATTCaaa. AGATCTPARTSEQUENCE 1 GGATCCaaa. CTCGAG AGATCTPARTSEQUENCE 1 GGATCC AGATCTPARTSEQUENCE 2 GGATCC Gly. Ser AGATCTPARTSEQUENCE 1 GGATCTPARTSEQUENCE 2 GGATCC

Type I Coding Sequences AGATCTATG_MIDDLE_OF_PART_TAAGGATCC AGATCTGTG_MIDDLE_OF_PART_TGAGGATCC ØThe start and stop codons are placed directly adjacent to the Bgl. II and Bam. HI sites, respectively ØStart codons are free to be ATG, CTG, TTG, or GTG

Coding Sequences Type II AGATCTATGAAATTTCCCGGGAAATTTGGATCC Type III AGATCTCATCATCATTAAGGATCC Type IV AGATCTAAATTTCCCGGGAAATTTCCCGGATCC ØCoding sequences allow the construction of ORF fusions for chimeric and tagged proteins. Gly. Ser scars separate junctions between fused peptides.

Ribosome Binding Sites AGATCTGAAAGAGGAGAAAGGATCC ØThe spacing of a ribosome binding site relative to the start codon is fixed. Shown is a (likely) strong RBS AGATCTATG_ORF_Part_TAAGGATCC. RBS. AGATCTGAAAGAGGAGAAAGGATCTATG_ORF_Part_TAAGGATCC

Promoters +1 | AGATCTTCC_Middle_of_Ptet_TAGAGATACTGAGCACGGATCC ØThe transcriptional start site (+1) is located at the position directly 5' to the Bam. HI site (whenever possible)

Terminators. . . CUUUCUGCGUUUAUA 3' | AGATCTCCA_Middle_of_Ptet_CTTTCTGCGTTTATAGGATCC ØThe transcriptional termination site is located at the position directly 5' to the Bam. HI site (whenever possible)