1、用snapgene进行多序列比对，导出alin文件
2、用python进行多序列比对美化
点击查看代码
from Bio import AlignIO
import os# ====== 用户参数 ======
alignment_file = "比对.fa" # 输入比对文件（fasta/clustal）
alignment_format = "fasta"
html_output = "msa_ruvc_all.html"# 背景渐变蓝色（保守性）
light_blue = "e6f3ff"
dark_blue = "08306b"# RUVC1/2/3 定义（基于 ungapped 序列位置，1-based）
RUVC1 = {"FnCas12a_6I1K_1": [(892, 953)],"LbCas12a_5ID6_1": [(809, 858)],"LbCas12a_6NME_1": [(808, 872)],"Lb2Cas12a_8I54_1": [(792, 852)],"ReChb_Cas12a_1": [(853, 914)],
}
RUVC2 = {"FnCas12a_6I1K_1": [(971, 1078)],"LbCas12a_5ID6_1": [(890, 1011)],"LbCas12a_6NME_1": [(890, 997)],"Lb2Cas12a_8I54_1": [(869, 992)],"ReChb_Cas12a_1": [(930, 1044)],
}
RUVC3 = {"FnCas12a_6I1K_1": [(1254, 1300)],"LbCas12a_5ID6_1": [(1138, 1228)],"LbCas12a_6NME_1": [(1179, 1228)],"Lb2Cas12a_8I54_1": [(1151, 1206)],"ReChb_Cas12a_1": [(1215, 1261)],
}# RUVC 样式
RUVC_color = "#FEBC28" # 所有 RUVC 使用同一颜色
RUVC_italic = False # 是否斜体# ====== 读取比对 ======
alignment = AlignIO.read(alignment_file, alignment_format)
seq_len = alignment.get_alignment_length()# 计算保守性
conservation = []
for i in range(seq_len):column = [rec.seq[i] for rec in alignment]chars = [aa for aa in column if aa != "-"]freq = max([chars.count(aa)/len(chars) for aa in set(chars)]) if chars else 0.0conservation.append(freq)# ====== 辅助函数 ======
def hex_to_rgb(hexstr):return int(hexstr[0:2], 16), int(hexstr[2:4], 16), int(hexstr[4:6], 16)def rgb_to_hex(r, g, b):return f"{r:02x}{g:02x}{b:02x}"lr, lg, lb = hex_to_rgb(light_blue)
dr, dg, db = hex_to_rgb(dark_blue)# 将 ungapped 座标映射到 alignment 座标
def build_ruvc_aligned(ruvc_dict, alignment):result = {}for rec in alignment:seq_id = rec.idseq = str(rec.seq)mapping = [i for i, ch in enumerate(seq) if ch != "-"]if seq_id in ruvc_dict:newranges = []seq_len = len(mapping)for s, e in ruvc_dict[seq_id]:if s > seq_len:continuestart_al = mapping[s-1]end_al = mapping[min(e, seq_len)-1]if start_al <= end_al:newranges.append((start_al, end_al))if newranges:result[seq_id] = newrangesreturn resultRUVC1_aligned = build_ruvc_aligned(RUVC1, alignment)
RUVC2_aligned = build_ruvc_aligned(RUVC2, alignment)
RUVC3_aligned = build_ruvc_aligned(RUVC3, alignment)# 判断某位置是否属于任意 RUVC
def in_ruvc(seq_id, pos, aa):if aa == "-":return Falsefor ruvc_map in [RUVC1_aligned, RUVC2_aligned, RUVC3_aligned]:for start, end in ruvc_map.get(seq_id, []):if start <= pos <= end:return Truereturn False# ====== 生成 HTML ======
with open(html_output, "w", encoding="utf-8") as out:out.write("<!doctype html><html lang='zh-CN'><head><meta charset='utf-8'>\n")out.write("<title>MSA - RUVC 高亮</title>\n")out.write("<style>\n")out.write("body{font-family: Consolas, monospace; padding:16px}\n")out.write("table { border-collapse: collapse; }\n")out.write("td.id { vertical-align: top; padding:4px 8px; white-space: nowrap; }\n")out.write("td.seq { vertical-align: top; padding:4px 8px; white-space: pre; }\n")out.write("span.res { display:inline-block; padding:0 1px; }\n")out.write("</style></head><body>\n")out.write(f"<h2>多序列比对（RUVC 高亮） — {os.path.basename(alignment_file)}</h2>\n")out.write("<div style='overflow-x:auto'><table>\n")for rec in alignment:seq_id = rec.idseq = str(rec.seq)out.write("<tr>")out.write(f"<td class='id'>{seq_id}</td>")out.write("<td class='seq'>")for i, aa in enumerate(seq):if aa == "-":bg = "#ffffff"color = "#000000"style_extra = ""else:# 背景渐变蓝色cons = conservation[i]r = int(round(lr + (dr - lr) * cons))g = int(round(lg + (dg - lg) * cons))b = int(round(lb + (db - lb) * cons))bg = "#" + rgb_to_hex(r, g, b)# RUVC 标注if in_ruvc(seq_id, i, aa):color = RUVC_colorstyle_extra = "font-style:italic;" if RUVC_italic else ""else:color = "#000000"style_extra = ""out.write(f"<span class='res' style='background-color:{bg};color:{color};{style_extra}'>{aa}</span>")out.write("</td></tr>\n")out.write("</table></div></body></html>\n")print(f"已生成：{html_output}")