Table 3.
Construct features
FLASH transgene featureSequence
attB–KpnI–promoter 5′-GATCTCTAGAGGTACCcgagattattgtttgtattgtgatgtttttttttaacgaatttaa-3′ 
V5–XhoI–AttB 5′-GGTAAGCCTATCCCTAACCCTCTCCTCGGTCTCGATTCTACGTAACTCGAGCCGCGGCCGCAGAT-3′ 
5′ UTR–EcoRI–coding 5′-GAAGAAACGTAAGCGATTGGTGAATTCCAAAATG-3′ 
EcoRI–FL 5′-GAATTCCAAAATGGAAACGCCTGCATATGCCAC-3′ 
EcoRI–aa 65 5′-GAATTCCAAAATGGACAGATCCCTTGAACTGGAC-3′ 
EcoRI–aa 78 5′-GAATTCCAAAATGGACGACTTTCAGAAGGCCGA-3′ 
aa 844–SacII–V5 5′-CTGCTGCgGACAAACCAATTCCGCGGTTCGAA-3′ GGTAAGCCTATCCCTAACCCTCTCCTCGGTCTCGATTCTACG-3′ 
aa 733–SacII–V5 5′-CTCACCCAGACTCCAAAACAGGCTCCGCGGTTCGGA-3′ GGTAAGCCTATCCCTAACCCTCTCCTCGGTCTCGATTCTACG-3′ 
FLASHmini junction 5′-CGAAAAGGACGATGT/ACCCAGACTCCAAAA-3′ 
FLASH transgene featureSequence
attB–KpnI–promoter 5′-GATCTCTAGAGGTACCcgagattattgtttgtattgtgatgtttttttttaacgaatttaa-3′ 
V5–XhoI–AttB 5′-GGTAAGCCTATCCCTAACCCTCTCCTCGGTCTCGATTCTACGTAACTCGAGCCGCGGCCGCAGAT-3′ 
5′ UTR–EcoRI–coding 5′-GAAGAAACGTAAGCGATTGGTGAATTCCAAAATG-3′ 
EcoRI–FL 5′-GAATTCCAAAATGGAAACGCCTGCATATGCCAC-3′ 
EcoRI–aa 65 5′-GAATTCCAAAATGGACAGATCCCTTGAACTGGAC-3′ 
EcoRI–aa 78 5′-GAATTCCAAAATGGACGACTTTCAGAAGGCCGA-3′ 
aa 844–SacII–V5 5′-CTGCTGCgGACAAACCAATTCCGCGGTTCGAA-3′ GGTAAGCCTATCCCTAACCCTCTCCTCGGTCTCGATTCTACG-3′ 
aa 733–SacII–V5 5′-CTCACCCAGACTCCAAAACAGGCTCCGCGGTTCGGA-3′ GGTAAGCCTATCCCTAACCCTCTCCTCGGTCTCGATTCTACG-3′ 
FLASHmini junction 5′-CGAAAAGGACGATGT/ACCCAGACTCCAAAA-3′ 

Engineered restriction sites are underlined, start and stop codons are bold, the FLASH promoter sequence is lowercase, the V5 tag is italicized, and “/” indicates the unique junction in the FLASHmini open reading frame.

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