Scoville Splice · ScovilleSplice.farm

SS-0094 — Collateral Damage

Pepper Heat Ladder Pepper · C. chinense · 10M SHU

Product Specification

Product ID
SS-0094
Product Name
Collateral Damage
Target Organism
Pepper (C. chinense)
Engineering Tier
Pepper
Series
Heat Ladder
Target SHU
10,000,000
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: Collateral Damage

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
312.5 μmol/g
Predicted DHC
187.5 μmol/g
Total Flux
3.9044 μmol/h
Vanillylamine Branch
0.523
Acyl Branch
0.385
Condensation Efficiency
0.6
Growth Impact
0.492 (fraction of WT)
Feasibility
EXPLORATORY

CRISPR Guide RNAs (POX Knockout)

TargetSpacer (20-mer)PAMGC%Efficiency
CaPOX1CCCGGGAAGCGGAGGGTCAANGG70.0%0.588
CaPOX2GTTAAGTAATCACTTGGAGGNGG40.0%0.928
CaPOX3CTTCGCCTAATTTATTATCANGG30.0%0.563
CaPOX4GACTTGACGGCTCTGCCGAANGG60.0%0.629
CaPOX5GGCCCCGTTAGGGTCGTAACNGG65.0%0.692

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)