VN550 pAC-cI_E34P-Asf1A_Term_f1-pOL2-62_L-eGFP-KanR

Plasmid Information

Purpose

Quantitative bacterial two-hybrid (qB2H). Expresses Asf1A.

Depositor

Oscar Ramos

Article

Guyot et al bioRxiv 2025.10.29.680610

Insert

Asf1A (ASF1A Human)

Sequence Information

Length

6,156 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

CDS 5

Label	Gene	Product	Position	Strand
CmR	cat	chloramphenicol acetyltransferase	0..6156	Fwd
DNA Sequence ATGGAGAAAAAAATCACTGGATATACCACCGTTGATATATCCCAATGGCATCGTAAAGAACATTTTGAGGCATTTCAGTCAGTTGCTCAATGTACCTATAACCAGACCGTTCAGCTGGATATTACGGCCTTTTTAAAGACCGTAAAGAAAAATAAGCACAAGTTTTATCCGGCCTTTATTCACATTCTTGCCCGCCTGATGAATGCTCATCCGGAATTCCGTATGGCAATGAAAGACGGTGAGCTGGTGATATGGGATAGTGTTCACCCTTGTTACACCGTTTTCCATGAGCAAACTGAAACGTTTTCATCGCTCTGGAGTGAATACCACGACGATTTCCGGCAGTTTCTACACATATATTCGCAAGATGTGGCGTGTTACGGTGAAAACCTGGCCTATTTCCCTAAAGGGTTTATTGAGAATATGTTTTTCGTCTCAGCCAATCCCTGGGTGAGTTTCACCAGTTTTGATTTAAACGTGGCCAATATGGACAACTTCTTCGCCCCCGTTTTCACCATGGGCAAATATTATACGCAAGGCGACAAGGTGCTGATGCCGCTGGCGATTCAGGTTCATCATGCCGTTTGTGATGGCTTCCATGTCGGCAGAATGCTTAATGAATTACAACAGTACTGCGATGAGTGGCAGGGCGGGGCGTAA Protein Sequence MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFLKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA
lambda repressor	cI	phage lambda repressor	1301..2012	Rev
DNA Sequence ATGAGCACAAAAAAGAAACCATTAACACAAGAGCAGCTTGAGGACGCACGTCGCCTTAAAGCAATTTATGAAAAAAAGAAAAATGAACTTGGCTTATCCCAGCCGTCTGTCGCAGACAAGATGGGGATGGGGCAGTCAGGCGTTGGTGCTTTATTTAATGGCATCAATGCATTAAATGCTTATAACGCCGCATTGCTTGCAAAAATTCTCAAAGTTAGCGTTGAAGAATTTAGCCCTTCAATCGCCAGAGAAATCTACGAGATGTATGAAGCGGTTAGTATGCAGCCGTCACTTAGAAGTGAGTATGAGTACCCTGTTTTTTCTCATGTTCAGGCAGGGATGTTCTCACCTGAGCTTAGAACCTTTACCAAAGGTGATGCGGAGAGATGGGTAAGCACAACCAAAAAAGCCAGTGATTCTGCATTCTGGCTTGAGGTTGAAGGTAATTCCATGACCGCACCAACAGGCTCCAAGCCAAGCTTTCCTGACGGAATGTTAATTCTCGTTGACCCTGAGCAGGCTGTTGAGCCAGGTGATTTCTGCATAGCCAGACTTGGGGGTGATGAGTTTACCTTCAAGAAACTGATCAGGGATAGCGGTCAGGTGTTTTTACAACCACTAAACCCACAGTACCCAATGATCCCATGCAATGAGAGTTGTTCCGTTGTGGGGAAAGTTATCGCTAGTCAGTGGCCTGAAGAGACGTTTGGC Protein Sequence MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQPSVADKMGMGQSGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEYPVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPDGMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSVVGKVIASQWPEETFG
S-Tag	—	affinity and epitope tag derived from p…	2239..2284	Rev
DNA Sequence AAAGAAACCGCTGCTGCGAAATTTGAACGCCAGCACATGGACTCG Protein Sequence KETAAAKFERQHMDS
EGFP	—	the original enhanced GFP (Yang et al.,…	2491..3208	Fwd
DNA Sequence ATGGTCAGTAAAGGCGAAGAGTTGTTTACGGGTGTAGTACCGATTTTAGTGGAACTCGATGGAGATGTGAATGGGCATAAATTTTCGGTTTCAGGTGAAGGTGAAGGAGATGCCACCTATGGCAAACTGACGCTGAAGTTCATCTGTACGACAGGCAAACTGCCTGTTCCGTGGCCAACACTGGTTACTACGCTGACCTATGGCGTTCAATGCTTTTCGCGTTATCCGGACCACATGAAACAGCATGACTTCTTTAAATCCGCGATGCCTGAAGGTTACGTACAAGAACGCACCATCTTCTTCAAAGATGACGGGAACTACAAAACCCGTGCAGAGGTCAAATTTGAGGGTGACACCCTTGTGAATCGGATTGAGCTGAAAGGCATTGACTTTAAAGAAGATGGCAACATTCTGGGTCATAAACTGGAGTACAACTACAACAGCCATAACGTGTATATCATGGCGGACAAACAGAAGAATGGGATCAAAGTGAATTTCAAGATTCGCCACAACATTGAAGATGGTAGTGTGCAGTTAGCCGACCACTATCAGCAGAATACCCCGATTGGCGATGGCCCAGTCTTGCTGCCGGATAATCATTACCTTAGCACCCAATCTGCGTTAAGCAAAGATCCCAACGAAAAGCGCGATCACATGGTCCTCCTGGAATTTGTGACTGCTGCAGGCATCACTTTGGGTATGGATGAACTGTATAAG Protein Sequence MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK
KanR	aph(3')-Ia	aminoglycoside phosphotransferase	3239..4055	Fwd
DNA Sequence ATGAGCCATATTCAACGGGAAACGTCTTGCTCTAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGGAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGTCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAACTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAA Protein Sequence MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF

promoter 2

Label	Gene	Product	Position	Strand
lac UV5 promoter	—	—	2056..2087	Rev
DNA Sequence TTTACACTTTATGCTTCCGGCTCGTATAATG
cat promoter	—	—	5834..5937	Fwd
DNA Sequence TGATCGGCACGTAAGAGGTTCCAACTTTCACCATAATGAAATAAGATCACTACCGGGCGTATTTTTTGAGTTATCGAGATTTTCAGGAGCTAAGGAAGCTAAA

protein_bind 1

Label	Gene	Product	Position	Strand
lac operator	—	—	2032..2049	Fwd
DNA Sequence TTGTTATCCGCTCACAA

rep_origin 1

Label	Gene	Product	Position	Strand
p15A ori	—	—	4763..5309	Fwd
DNA Sequence TTGAGATCGTTTTGGTCTGCGCGTAATCTCTTGCTCTGAAAACGAAAAAACCGCCTTGCAGGGCGGTTTTTCGAAGGTTCTCTGAGCTACCAACTCTTTGAACCGAGGTAACTGGCTTGGAGGAGCGCAGTCACCAAAACTTGTCCTTTCAGTTTAGCCTTAACCGGCGCATGACTTCAAGACTAACTCCTCTAAATCAATTACCAGTGGCTGCTGCCAGTGGTGCTTTTGCATGTCTTTCCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGACTGAACGGGGGGTTCGTGCATACAGTCCAGCTTGGAGCGAACTGCCTACCCGGAACTGAGTGTCAGGCGTGGAATGAGACAAACGCGGCCATAACAGCGGAATGACACCGGTAAACCGAAAGGCAGGAACAGGAGAGCGCACGAGGGAGCCGCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCACTGATTTGAGCGTCAGATTTCGTGATGCTTGTCAGGGGGGCGGAGCCTATGGAAA

terminator 1

Label	Gene	Product	Position	Strand
T7Te terminator	—	—	4666..4694	Rev
DNA Sequence GGCTCACCTTCGGGTGGGCCTTTCTGCG