向 NCBI 提交基因簇的时候需要提供 sqn 格式的文件，这个文件需要通过 tbl2asn 生成。

# 文件准备

tbl2asn 依赖三个文件来生成 sqn 文件：

文件 1：fasta 格式的基因组序列文件

Header 处的中括号部分可以不写。

	>Toyoncin_biosynthesis_gene_cluster [organism=Bacillus toyonensis] [strain=XIN-YC13] [topology=linear] [moltype=DNA] [tech=wgs] [gcode=11] [country=China] Bacillus toyonensis strain XIN-YC13 Toyoncin biosynthesis gene cluster, complete sequence
	ttaaaa taatttaata
	gggaagtttt ttagttgttt tggactcttc ccaaacactg ctttaagtgt tggattaaca
	tcatccctat tccccgaaaa cataatgtga ggatttatga ataatgcata tgctctaaca
	ttattatcat caacaccact ctctgaacga gccataatac ccttatcaat taattttcta
	accaatggac taactttagt ttcatgtctt ccaatttttt tagctaattc tcgctgagtt
	aatgggattt gttctttcga attaatatca ttaactaaac aattacttaa aaaacctaca
	cacattgaaa tatctactaa aaataccttc tcagcatttg ttaaataatc aatttcgaac
	aaatactgga tattttgttg aataatctga acaaacttcg ctttattttt cactttacgc
	tcaggaacta atttcattcc tcttgaacga gcttttgatt gaagtttatt tgctaaatac
	atctcttctt cagacaagac ctttaaatcc tcaatatctc tcaatcttga atttttttca
	gcttgttcta agttgataaa ctttgacata ttctttttgc tcctcttttc taagattttc
	aactagagaa ggaaaaaatt ttatgttatg attcctgtag aatttacaat tcaatatgta
	caaaagaact ccccttttct aattgatagt ttggtcgctt tcaattataa tacaagggga
	ttttttacat cttaaaattt ttcatttttg aatcaatccc tgaaaatata aagaacacat
	cacataaatt attcttaata ttttataatc gaaaaaataa taggaataaa gaaaaatact
	gcaataaata tattcatctg tttcttactc aaaccggcca ctatatttaa tcccattcct
	ataataatta attcccaaat tgaaaacact tcaaatttac tacaaattat atatagtaat
	gtacctggtt caaatattga acccaaacta gtatacgtta ctatttctcc tcctataaat
	agtgttaata atgtattaat taatttacct aaaatagaaa ttacactagc aaatattgta
	atagatacta actttttata agaaacatct ttactcatca gcatcattac aatctttaaa
	ataatccccc aaataaaagg tgtaattaaa gcaatgaaaa tcgatgcaaa acctcctaac
	atcatttggg aaacaagggg tatttccata tctgcaaata cttctttttg aattttaacc
	aattctggat tgctatgtct tgcatataca gataaaatcc ctattattgc ttgtataact
	gataaataca taagaggaaa ccatatcgga ctaattattt tcatacgctc gaattcagaa
	ataggagatg taatcataaa aattagagat ggtttttcat aattgttttt ttctttattc
	actactaaac tattatccat atattaacac cttctttttt tattcataac gtaatgcttc
	aattggatct aattttgcag ccttattggc tggaatcaat ccaaatataa taccaagcga
	catcgaaaat aatacgccac ccacaacaac ttcccatgaa acaagaggcg gccattttgc
	aaatgtggac acaatgtacg ctccacaata accaagtcca atcccaatca atccaccaag
	aagtgtcaac ataattgctt caattaaaaa ttgcaacaaa attttaccac gcgttgctcc
	aagtgcttta cgtaccccaa tctcacgtgt acgctctgtt acagaaacaa gcatgatatt
	cataactcca attccgccta caactaaaga aatacttgca atacctgcaa taatcattgt
	cataatatta gtaactttag aaataccttt ttggatttct tctaaattta caatttcata
	tttcccttta aactcttcag attgtctatc atttaataat tttactccct tttttccagc
	tgtttgtaat tgatcaaccc ctattgcttg aattgtaata gattgttgag agttatcatc
	tccatataat attggccata ttgaaagtgg tattaaaatt tctgacattc caaaaccaag
	ctcttcatct cctgaactga atagaccaat aatttgaagt ggctgacctt taatttctat
	aattttacca atgactgatt catgctcatt aggaaataac tctttcacta atgtttgatt
	aaccattatt acattattac cttgcatcaa atcatcttca ttaagagaac gacctttctc
	tattttcatt ttagtcatat taaaatattc ttttgtaata ccatttatat tagttacaac
	ctttttatca tcaccaatta atgtctctgt actagagttt tgaacaatta catttttaat
	ttcttttatc ttttttaact caaaaagatc ttcttcactt acagatggtt ttttgtcatt
	catagatcct gttgttaata actcattaat atcttcttta tatgtaatcg gaatagtgtt
	attgccagaa gcggtaaatt gtgatttaag cattgcttct ccacctttac caatggctac
	aacagtaata atagaaccta caccaataat aattccaagc atcgtaagag ctgagcgcag
	tttatgagct aaaatagaag ataaggcaat ttttatacta tctaataaac tcataccata
	caccttctat cttctgtaat tttcccatct cgcaatatga tgcgacgtga agaataagct
	gctacctctt cttcatgtgt aaccataacg attgtcgtac cttctgcatt taacttcgta
	aagatatcca taacttgtgc accagacttc gtatcaagcg caccagttgg ctcatcagcc
	ataataaacg ttggattatt cgcaatcgat cttgcaatag caacacgctg cttctgtcca
	cctgacagct cactaggtaa atgatgtact ctatccgcta atccaacttt cccaagcgct
	tcgagcgctc tttgacgacg ctctgctttc ttcactccac cataaatcag tggtaattca
	acgttttcca ctgcggaaag gcgcggcaat aaattaaaat gctggaacac aaaaccgata
	tattcattac gaattaaagc aagttttgac tcatctgctg ttaaaatatt cacatcattc
	agcatatatt cgccttctgt tggacgatct aaacaaccga taatattcat aagagttgat
	ttaccagaac cagacggtcc cataattgaa acaaattcac caccttgaat agttaaacta
	ataccgtgca aaataggaac cgccattttt ccttgataat acgttttagc aatattattt
	aacgtaatca tttctctttc acttccattc cgtcatatac gttgtcggaa ggatttttaa
	ccaccttttg ccccactgtt gcgccctcta caatctctgt ccaatctcca tcagtagcac
	cttttttcac attttgttta cgaagcttac ctttctcttc gatatataca aatgcatcat
	cgcctttttc aacaatactc ttacttggaa cagcaatcat tcttttattc tctaaattta
	cttgtaacga aacatgataa cctggagata aaccatcttg actatcaaga cttgctttat
	atgtatattg agacatattt tgagtcactt cccccatgcc atcagcttga gccatttcta
	cacttgttgg gaactcactt acctctgtaa tcttccctgt ccactttttc ttactatttg
	ctttcgcagt tacagtaaac gtttgatcct tttgaatttg cgacttctga agctcagtta
	atgttccttg aatttggaat ggatctttag aagcaacttg taaaaaggct ttcccttgac
	cacctaacgc ttgtgatgaa ctttgtgctg catctttatc taacttttga acaacaccag
	caaaattgct ataaatcgta agttcgttct gctttttatt taactcttct ttttgtaact
	tccctttctc tttctcaagg tctgttgtct tttgcgctat ttctaattca cttacttgct
	cttccatcgg atctattact tctttcccag ctccgctatc tttcgccttc ttaatttctt
	tcttcaacga atcaatcttc tttttccctt ggtcataacg catatctgcc atcttttgat
	caagcacagc ttgcttcatt tgcaaattaa tttcttcatt atcgtaagaa aacaatttcg
	ttcccttttc tatttcttgt ccttctttca cttcaatatc tttcactttt cctttagtca
	gatccgcgta gaaactttca atattccccg gcttcacttg accagaaatt aactttgtat
	tattaagatt gcgctctgtg actttttcaa aactaacagt atctattttt gttaccgctt
	tcttcttact ttgcactacg aaaatattaa taaatgtaac aataacaatt aacgcaataa
	ctccaataat agctcctttc tttttatttt taaaaataaa aagttctttt ttaatcacaa
	caatcttctc cttattcata tctaaaattt aaacttttaa attttacata aaaatttaaa
	acttctaaaa tataacatgt ataatttacc atagatgatt tattttgtat aatataaaaa
	tatctatata aataatgcta attttcaaac aatggggtgg aagatactaa tgttagaaaa
	aaaagataga ctaacagaaa tagaggaaca aattatatac ttaatttcaa aggaattagg
	aaataaagaa atagcggaaa aattaaatta ttcacaacgt agcatcggtt acaaaataaa
	taatattttt aaaaaattaa atgttaattc aagaatcgga ctgattatag aagctgtaaa
	aaaaaatata atttaaatat aagaatgctt tcatgttaat attttataga aactaaatat
	agaggtgatt aaaatgcaaa aattttttga agctattagt gctataggta tagtaggtta
	ctttttaggt aaattcacaa gtattccttt aatagacaaa tatacattgt atttcggcgt
	aatgttgatg attggggtta ttggaagatt tattataaaa gtaattaact cagaagaaga
	gacacatgat tcaaacaaat aaaatactct aataaaaatg gaagaagatt gcacttaagt
	gcaatcttct tccattttta ttgaaaattg attaaataat gttaatattg caattgtgtg
	gtgcagatta gggtgattat gtaatagggg gaaattaaaa atgatcaata cagcttggaa
	aattattaaa gcactacaaa aatacggtac aaaagcatac aatgttatca aaaaaggcgg
	ccaagcaatg tacgacagct tcatggcagc taaagctaaa ggttggacac atgcagcttg
	gtggctagta gaacatggtt caactttagg aacattctat gatttattaa aagctgctgg
	attaatcgac taattacagc aactaaacaa ctaaacaact aaacaactta aaaatacaaa
	ttaccctaaa ctgtacccct attacatatt aactaattat tttaaaggtt ggatgataat
	atgtcaaata acatcatatc tgtaaaaaat ttaattaaaa gcttcgataa caaaatagta
	ttagataaat taaatttcga aatgaaagaa aactccactg ttgtaataat aggtaaaaac
	ggtgcaggta aaagtgtctt tctaaattgt ttacttggat ttattcatta caaccaaggt
	tcaatactaa tagatggaca acctgtagaa aatcgattac atctccgcaa gattacatcg
	ttaatttctt cagaccatca agaacatcta aatttattaa cccccaatga atatttttct
	tttttacaag atatttacca actaaaaagt aataataaag acaaaattca aaattactca
	gaagatctat atgttactaa agaactcaat actgtatttt catcactttc ttttggaaca
	aaaaagaaaa tacaattaat tggtagccta ttatattctc ctaaattatt gatttgcgac
	gaaatatttg aagggcttga tacagactca gtaaaatggg ttaaaaactt atttcaacaa
	agaaaacaag aaaatctttc tactttattt acaactcata ttactgaaca tataacagat
	ataacagaaa aaaattacat acttgaaaat ggaaaattaa ttgtgtaagt ttaaccactt
	atatttaaag ctaaaattaa ggagcttaaa atatgaattt taatatatat aagagactat
	atgataaatc aacagaagaa aaaagcaaaa caataacaaa acaaatatta tttggaatta
	taaatagttc tatattaata ggtatactac tcacatgttt ggagattttc aactttaaaa
	tttcaactgt aatgtatggt tatttcacta tatatataat actagaactt ttactattat
	tctctgcaaa tcaactatat gaaagtacag aattcataat aaaattcctt aaatatacac
	caataaccat aaataaacta tatttctcac attttctaag ttctaaatat tcattttcca
	atctttttga aataataact ctcacatcaa ttttattaat atataatgtc gatatcttat
	attcatttat tttcataatt agcttacaaa ttattagctt aataagaaca tatttagaat
	ttttactatt atattctcaa aaaaaacagg ttaaaatttt tactctaacc cattttgttt
	tcataatatc tatggttttt tatattattg ttaaaacaaa atcgatagat ttagtattct
	ttgaaaacac aaatatgtta attatatctg ttcttctcat aacattcttg atatcacttt
	taacatataa acatattata gaatacttaa tgaaaaataa tgaaattgta tataatgcta
	tttttatcaa gttaactttt aacacagcta atttaattag taaattattt aaatttaata
	catcaattgc atctttaata aaaatacata taatacgatt attacgtaat caagactata
	taagtagatt actaaaaata ggaatattac tatttatttt ttcttctata agctttctat
	ttttcgataa atcatcaaca aacaatgaaa tgagtgatat actttacttt tcatttttta
	tttccttatt tagtttttct aacatacgat tagactataa cttagtttct aaattaagct
	tagaggatta tccaataaca aaattacaat caagattaag cattgatata gcacatggaa
	ttttactatt tatactatct ttatttcttt tattaacaca atacttattg aatccaacaa
	atattctaac tctaattgat ggtttattat catttatttg tttttatttt ctaagtcttg
	gtatagaaaa agcagatatt ataataacac caaaaacaaa atggaaaatg tatccattat
	tttttgtgat gggattaata attgaagcaa tatttctatt aaaattcaaa atatggataa
	aattaataac tttattcctt tgtatactgt ggtcatattt acgtgtttat tggaaattaa
	aaaaacaata aacacaatta aaaagttccc ttcatatttt ttgaagggaa cttttatttt
	aaacaaaaat tacaaacaag caaagttatt taaaagtaaa cttttaaaat tattgaatta
	ataacaatta gtctaagata tatcagccaa atttaatttt taaacaaacc gaaaaaccct
	ttccgttttt gtttctgatt ttggctctgt atttctctaa tgttttcaag caataactga
	tctcgttttt caaatttttt ctctataaaa acctctaatt caatattttt atcttctact
	tcctttaatt ttctctccgt attagccaaa tgttcttttg tggtaactaa ttcattcgta
	atctcttgta atttttgaac aagcgtttga ttgaactgat tttgtaattg ttgattttct
	aatacttcat ccaacttctt ttctaattcc gatttttcct ctcttgaaac aaacaaatca
	agttctccat tccgccatgc tcgaacttga tcataagacc atttttgcac ctgtattttt
	tctttaattg ctatcaattc ttctatattt tcctttgagt acctacgatg ccctccctga
	cttcgctccg tttgtatatt aaattcgttt gaccatgctt ttaacaagtc aggggtaatc
	cctaaacgat ccgcaacaat tttcggtgta tacatttctg attttaattc caa

文件 2：描述基因特征的 feature table 文件（.tbl）

该文件可以用 prokka 对文件 1 进行注释而得到，但是需要自己加以修改，加上文件前几行以及 gene 相关的信息，各列之间用制表符分隔。

	>Feature Toyoncin_biosynthesis_gene_cluster
	1 8409 source
	organism Bacillus toyonensis
	mol_type genomic DNA
	strain XIN-YC13
	585 1 gene
	gene orf1
	585 1 CDS
	inference ab initio prediction:Prodigal:002006
	locus_tag Toyoncin_biosynthesis_gene_cluster_00001
	product MarR family transcriptional regulator
	1476 811 gene
	gene orf2
	1476 811 CDS
	inference ab initio prediction:Prodigal:002006
	locus_tag Toyoncin_biosynthesis_gene_cluster_00002
	product YIP1 family membrane protein
	2710 1496 gene
	gene orf3
	2710 1496 CDS
	inference ab initio prediction:Prodigal:002006
	locus_tag Toyoncin_biosynthesis_gene_cluster_00003
	product ABC transporter permease
	3387 2707 gene
	gene orf4
	3387 2707 CDS
	inference ab initio prediction:Prodigal:002006
	locus_tag Toyoncin_biosynthesis_gene_cluster_00004
	product ABC transporter ATP-binding protein
	4595 3384 gene
	gene orf5
	4595 3384 CDS
	inference ab initio prediction:Prodigal:002006
	locus_tag Toyoncin_biosynthesis_gene_cluster_00005
	product RND family efflux transporter, MFP subunit
	4746 4952 gene
	gene orf6
	4746 4952 CDS
	inference ab initio prediction:Prodigal:002006
	locus_tag Toyoncin_biosynthesis_gene_cluster_00006
	product Helix-turn-helix transcriptional regulator
	5010 5198 gene
	gene orf7
	5010 5198 CDS
	inference ab initio prediction:Prodigal:002006
	locus_tag Toyoncin_biosynthesis_gene_cluster_00007
	product Putative membrane protein
	5337 5549 gene
	gene toyA
	5337 5549 CDS
	inference ab initio prediction:Prodigal:002006
	locus_tag Toyoncin_biosynthesis_gene_cluster_00008
	product Toyonsin precusor
	5657 6304 gene
	gene orf9
	5657 6304 CDS
	inference ab initio prediction:Prodigal:002006
	locus_tag Toyoncin_biosynthesis_gene_cluster_00009
	product ABC transporter ATP-binding protein
	6349 7707 gene
	gene orf10
	6349 7707 CDS
	inference ab initio prediction:Prodigal:002006
	locus_tag Toyoncin_biosynthesis_gene_cluster_00010
	product Putative membrane protein
	8391 7849 gene
	gene orf11
	8391 7849 CDS
	inference ab initio prediction:Prodigal:002006
	locus_tag Toyoncin_biosynthesis_gene_cluster_00011
	product MarR family transcriptional regulator

文件 3：描述作者信息的模板文件（.sbt）

可以在 NCBI 上生成该文件。

	Submit-block ::= {
	contact {
	contact {
	name name {
	last "xin",
	first "bingyue",
	middle "",
	initials "",
	suffix "",
	title ""
	},
	affil std {
	affil "Huaibei Normal University",
	div "College of Life Sciences",
	city "Huaibei",
	sub "Anhui",
	country "China",
	street "Dongshan road No.100",
	email "xinbingyuex@163.com",
	postal-code "235000"
	}
	}
	},
	cit {
	authors {
	names std {
	{
	name name {
	last "Xin",
	first "Bingyue",
	middle "",
	initials "",
	suffix "",
	title ""
	}
	}
	},
	affil std {
	affil "Huaibei Normal University",
	div "College of Life Sciences",
	city "Huaibei",
	sub "Anhui",
	country "China",
	street "Dongshan road No.100",
	postal-code "235000"
	}
	}
	},
	subtype new
	}
	Seqdesc ::= pub {
	pub {
	gen {
	cit "unpublished",
	authors {
	names std {
	{
	name name {
	last "Xin",
	first "Bingyue",
	middle "",
	initials "",
	suffix "",
	title ""
	}
	}
	}
	},
	title "Purification and characterization of a novel leaderless bacteriocin, toyoncin, produced by Bacillus toyonensis XIN-YC13 that specifically active against Bacilus cereus and Listeria monocytogenes"
	}
	}
	}
	Seqdesc ::= user {
	type str "Submission",
	data {
	{
	label str "AdditionalComment",
	data str "ALT EMAIL:xinbingyuex@163.com"
	}
	}
	}
	Seqdesc ::= user {
	type str "Submission",
	data {
	{
	label str "AdditionalComment",
	data str "Submission Title:None"
	}
	}
	}

注意：文件 1 和文件 2 的序列描述信息必须一致，此例中均为 “Toyoncin_biosynthesis_gene_cluster”。

# 文件生成

tbl2asn -t template.sbt -p ./ -V vb -x .fna

-t 模板文件
- p 输入文件所在路径
- V
-v 生成验证文件，保存错误信息
- b 生成 gbf 文件
- x 文件 1（FASTA 文件）的后缀名，根据实际情况填写

注意：如果用 Prokka 带的 tbl2asn，生成的 sqn 和 gbf 文件中的日期通常是 1-JAN-2019，需要自己手动改正为当前时间，这是因为 Prokka 里的 tbl2asn 是经过修改的。建议使用官方版的 tbl2asn，可避免日期错误。

# 参考

tbl2asn
上传基因组数据到 NCBI

NCBI 序列处理生信软件

# 文件准备

# 文件生成

# 参考

R语言安装依赖包错误集锦

利用NCycDB数据库从宏基因组中预测氮循环基因