Scoville Splice · ScovilleSplice.farm

SS-0095 — Extinction Protocol

Pepper Heat Ladder Pepper · C. chinense · 12M SHU

Product Specification

Product ID
SS-0095
Product Name
Extinction Protocol
Target Organism
Pepper (C. chinense)
Engineering Tier
Pepper
Series
Heat Ladder
Target SHU
12,000,000
Pun1/CS Variant
Triple
Gene Count
10
Construct Length
15,914 bp
Construct Type
T-DNA binary vector
Selection Marker
nptII (kanamycin resistance)
Promoter System
2A11
Codon Optimization
pepper
Capsaicinoid Profile
extreme multi
Burn Character
Heat Ladder: Extinction Protocol

Gene Cassette Map

LB — [nptII] — [pAMT] — [Pun1/CS Triple] — [BCAT] — [KAS] — [FAR] — [PAL] — [C4H] — [4CL] — [HCT] — [COMT] — RB
GeneFunctionRole
pAMTPutative aminotransferaseVanillylamine biosynthesis
Pun1/CS (Triple)Capsaicin synthaseAcyl-vanillylamine condensation
BCATPathway enzymeCapsaicinoid pathway
KASPathway enzymeCapsaicinoid pathway
FARPathway enzymeCapsaicinoid pathway
PALPathway enzymeCapsaicinoid pathway
C4HPathway enzymeCapsaicinoid pathway
4CLPathway enzymeCapsaicinoid pathway
HCTPathway enzymeCapsaicinoid pathway
COMTPathway enzymeCapsaicinoid pathway

Flux Balance Analysis

Predicted Capsaicin
300.0 μmol/g
Predicted DHC
270.0 μmol/g
Total Flux
0.3748 μmol/h
Vanillylamine Branch
0.62
Acyl Branch
0.615
Condensation Efficiency
0.435
Growth Impact
0.437 (fraction of WT)
Feasibility
EXPLORATORY

CRISPR Guide RNAs (POX Knockout)

TargetSpacer (20-mer)PAMGC%Efficiency
CaPOX1ATGTCTACGGCGCAGCAAGCNGG60.0%0.657
CaPOX2TGCCGGTATAGAGTTGATCANGG45.0%0.949
CaPOX3CAACCCCTCACGTTGATTACNGG50.0%0.584
CaPOX4ACCCCTATTAGATTGAAGTGNGG40.0%0.783
CaPOX5ACGTGAGAAACAAGAGATTGNGG40.0%0.729

Transformation Protocol Summary

Vector
T-DNA binary vector in Agrobacterium tumefaciens GV3101
Selection
50 mg/L kanamycin
Confirmation
PCR with construct-specific primers + HPLC capsaicinoid assay
Expected Timeline
4-6 weeks (callus to T0 plant)