Scoville Splice · ScovilleSplice.farm

SS-0091 — System Shock

Pepper Heat Ladder Pepper · C. chinense · 7M SHU

Product Specification

Product ID
SS-0091
Product Name
System Shock
Target Organism
Pepper (C. chinense)
Engineering Tier
Pepper
Series
Heat Ladder
Target SHU
7,000,000
Pun1/CS Variant
Triple
Gene Count
9
Construct Length
14,655 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: System Shock

Gene Cassette Map

LB — [nptII] — [pAMT] — [Pun1/CS Triple] — [BCAT] — [KAS] — [FAR] — [PAL] — [C4H] — [4CL] — [HCT] — 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

Flux Balance Analysis

Predicted Capsaicin
284.375 μmol/g
Predicted DHC
91.875 μmol/g
Total Flux
2.0191 μmol/h
Vanillylamine Branch
0.84
Acyl Branch
0.261
Condensation Efficiency
0.635
Growth Impact
0.452 (fraction of WT)
Feasibility
EXPLORATORY

CRISPR Guide RNAs (POX Knockout)

TargetSpacer (20-mer)PAMGC%Efficiency
CaPOX1TAACTTCGCCATAGTGATAGNGG40.0%0.9
CaPOX2ACTTTGTCCTTATCGCAGAGNGG45.0%0.551
CaPOX3TTGGAGAGACCCAAACTGGCNGG55.0%0.802
CaPOX4CATTGGCAGCCAGCTTATGANGG50.0%0.565
CaPOX5GGAGCTTCTAGAATTTAATCNGG35.0%0.665

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)