| 质粒名称: | pL99 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 78577 |
| 存储实验室: | Xuming Mao |
|
载体骨架基本信息 |
|
| 载体名称: | pIJ8668 |
| 载体出品公司: | -- |
| 载体抗性: | 安普霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 有关克隆的详细信息，请参阅出版物 |
| 载体类型: | 细菌表达；链霉菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | 诱导型PnitA-NitR系统 |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | 5'-gcatgctaaaccgatacaatt-3' |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pL99 was a gift from Xuming Mao (Addgene plasmid # 78577 ; http://n2t.net/addgene:78577 ; RRID:Addgene_78577) |
| 参考文献部分: | Construction of over-expression shuttle vectors in Streptomyces. Sun N, Wang Z-B, Wu H-P, Mao X-M and Li Y-Q. Annals of Microbiology Dec 2012, Volume 62, Issue 4, pp 1541-1546 10.1007/s13213-011-0408-1 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pL99 7554 bp ds-DNA circular SYN 12-NOV-2019
DEFINITION Over-expression shuttle vector for gene cloning and transfer from E.
coli to Streptomyces. Contains inducible PnitA-NitR system .
ACCESSION .
VERSION .
KEYWORDS pL99
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7554)
AUTHORS Sun N, Wang Z-B, Wu H-P, Mao X-M, Li Y-Q
TITLE Construction of over-expression shuttle vectors in Streptomyces
JOURNAL Annals of Microbiology Dec 2012, Volume 62, Issue 4, pp 1541-1546
REFERENCE 2 (bases 1 to 7554)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 12, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7554
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 55..94
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
terminator 1177..1216
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
primer_bind complement(4297..4313)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4297..4313)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4310..4332)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4321..4337
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4345..4375)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4390..4411
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4528..4545)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4699..5287)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4779..4798)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS 5491..6291
/codon_start=1
/gene="aac(3)IV"
/product="aminoglycoside 3-N-acetyltransferase type IV
"
/label=ApmR
/note="confers resistance to apramycin (Paget and Davies,
1996)"
/translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
CDS complement(6725..7096)
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT 7129..7238
/note="incP origin of transfer"
terminator 7402..7441
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
ORIGIN
1 catatgctgg acggcggatc cgccgatatc gccaagcttc agagatctta aaccgataca
61 attaaaggct ccttttggag cctttttttt tggaggcgaa ctcccttatg cgggtggcgc
121 agaatgccag gacccttgtc attccacgtc aattcatgcg ccttttcacc tcgtactgtc
181 ctgccaaaca caagcaacgg aggtacggac atgaacacct tcttctcctc cgaccaggtc
241 tcggcgcccg accgcgtcgc gctctggcac gacgtcatct gccgcagcta cgtcccgctg
301 aacatcaccc tgaccagcga gcagcccttc atcggcaccg tctccaccgg caacctgggc
361 accgtccgca tcgccacgtc ctcgtccctg ccccagcaga tcacccgcac ccgccgcctg
421 atccgccagg acgagcgcga gtacctcatg gtcggcgtgc agtccgccgg ccacgccctc
481 gtgcagcagc acggccgcac cgcccgcgtc ggccgcggcg gcctggtctt ctgggacacc
541 cgccacccgt acgacatcct cttcccgacc gactggcgca tgagcgtctt ccagttcccg
601 cgctactcct tcggcttcac cgaggacttc atcggccgca tgaccgccgt caacgtcggc
661 ggcgaccgcg gcatcggccg cgtggtctcc tccttcatga ccagcatcaa cgacgcgacc
721 gacgccggcg acctggcgga ggtcgcctcc ctccacaaca gcgccgtcga cctgctgtcc
781 gcggcgatcc ggaccgagct ggccgaccag gccgccgcct ccgacggcct gctcgagtgc
841 gtgctcgcct acatccgcca gaacctggcc gacccgaacc tgtgcgcctc ccagatcgcc
901 gccgagcaca acgtctccgt gcggaccctc caccgcctgt tctcggccac cggccagggc
961 gtggccgagc acatccgcaa cctccgcctc gagcgcatca agaccgagct ggccgacccg
1021 accagccggc gctacaccat cagcgccctg gcgcgcaagt ggggcttcct cgacccgtcc
1081 accttctccc gcgcgttcaa ggacgcctac ggcatcaccg cccgcgagtg ggcggcctcc
1141 gcctccgcct ccccgaccga ggtctcgtag taaaccgata caattaaagg ctccttttgg
1201 agcctttttt tttggagtct agactgcagc cgggggtccg gcagggccgg agcgggcgga
1261 accgtgctct gacctgcggc ccgagtttcg tcacgtgacg gaatggaagg ctgctgcatt
1321 tcgtcacgtg acgtatctcg gcgagcgact gccgacgcca cggcggacac gatcgcctcg
1381 cgctggcgcc gggcctcgta cgcccgctgg cggcaggagc ggcggcagta gtcccggctc
1441 cggccgacgc cggattgctt gatctccgag ccgcaccagg cgcagagctt cgcgccgtcg
1501 gcgtccctgg gggtggtggt gctcatggcc gacgaccgta cgcggcacgt ctcgtagcga
1561 ggcgagtcgg gcgcgaggta ccgcctgcac gaagtgccgg cggggccgac cccgggcgag
1621 taatcccagg attactcccg cggcttcgac cccggccgcc gtcgccgcgt acgtcaccga
1681 cccccgccgt acgtcaccgg gatgacgtac ggcggggggg agcgagttag tgcgaagtgg
1741 gcccacttgc gagccgggcg atgtgccggg cggcccgctc ctggcggtcg tcggcgtcgt
1801 cgtcctggtc gtcgtcctgc tctcgccgtc ggcgtgcagt tgcttcctcg cggcgctggg
1861 cgagggcggc gagcatgtcg gcgtacgcct cggccacctc ccccgccgtg agcaccacca
1921 ctgtgtcggc cgcgtcggcc agcgccagga cctcccgcac ccgttcgccc acggccgccg
1981 aatcctcgtt gccgtccttg ccttcggcgg cccgggtcgc ctcgaggtcg agggcgcggc
2041 gggtgaccgc gtgccatccg tcctcggtca cggcgacccc ggcccgcagc tccccgccgt
2101 cggcgtcggc cgccaggagc agatcgaggt cgtcggcctc ggtgtcgccg ccgtcgagcc
2161 cgagcatctg ccgcaggtag cgggtccatt cgatggcccg gcgtccccgg gttgcccgct
2221 cgtactcgtg ccagcgcgag aggttccact ccagcgagcc gaccccggcg gcgtcgtcct
2281 cggtcatgcc gccggtcagg tccccgatcc gtccgaggag ttcgaacggg gcgacgttcc
2341 cgccggtcgc cgtcttgagg tcggcgcggg cgagttcgag ggcgggcgcc ttcccgtcct
2401 gggtcttggc gatgtactcg gcgaggtcgt tggcgtcgcg ctcggtctcc agccgcttga
2461 agtcgacgcc gtgccggtcg tcgggcgtga aggcggggtt gaccttgcgc agggcggcgg
2521 tccacacgga ccgccagtgc ccctgccact cgtcgagcgc ggcgccggtc ggctcgaagg
2581 tggcgacgat ctgcttcgcg gaccgctccc cctcggtccg gccgccgacc aggacgatcg
2641 cgtggatgtg cgggtgccag ccgttgatct gccccacggt gacttcggtc gcgcggatca
2701 tgccgacgta cccgatccgg tctcggatgc cctcgcggtc ggcggcccgg tgcccgtcct
2761 tggcccggcg tccggcccac gtgccgcccg tgatcagtcg ctggtaggcg cccggccgcc
2821 gggggctgtc cggcgtcttc cgggtgccct ggagggcgtc catgaggtcc gcgagccggt
2881 ccgtgtgccc atggcgggcc gtgaaggtga ccaggtaggc ggtccccccg cgcttgatcc
2941 actcgaccac ggcggcggtg atctcctcgg cccgcttgtg ccggatcgtg gcggcgcaga
3001 ccgggcagag ccagatccgc ccgcaccgca tcaggcccag gaccacggac gttccggccg
3061 ccgtctgggc gacgattacg ccggaggcag ggtccatcag ggcgcggccg cagcccttgc
3121 acgcggcgtc cccgctgatc cgccacagcg tccggcggcg gctgtaccgg gcggctttcc
3181 gcagtcgggc agcgtcgctc cgcgacgtgc ttcctacttc cgagaggctg tcgcctctcg
3241 ggctctcccc atccaccccg tccggagaaa ccgcaggtcg gaggggtgcg ggaaactctg
3301 ttgtttcttt cccaaggtgt tcgcttttgc ctcgggcggc atctcgcgtc acacgcgcga
3361 tcgcccgctt cgctgccatc cggcagcggt ctgagcagta gatacgcggc cgtttgcccg
3421 gtgtgtgggc aattgcggtc ccgcagtggc agcggggccc ggcgggccga tctggcaatg
3481 cctcggcatc gctccgtact ctgggcacga gcaacgttcc tgtctcgccc ggctaagggg
3541 cgcgagtctg ggagcggacg ggtcggaggt gcgaagtccg gcccgttgct ctttggtctg
3601 gtgggaatcc tggcaccaat cgggccagag gttccctccg ccactcccga cgccccttgg
3661 ggctggtgtg acttggaggg ccgaagagag ccccgccggt gatccggcgg ggctttgacg
3721 tgcggtcagt gcgtgtgtcg gcgagcgatg gccacgaggc cctggaagcc gagcggtccg
3781 gcgaagtcgg cccagtcgca accgggctca gcgcagtggg cggaccagcc accgccgttg
3841 gggtcctgga ccaggttcac ggtcccctcg gtcaggcgtc cgtcgaagtc ggtcatggtc
3901 ggtctcctgg tgggtggggg cggggcgcca gcacgaagtg ccggcgcccc gcgggggttg
3961 gtcgggtcag gcgccgaacc ggcgggcggc ggcggcgacc aggccgtcgg cggcggccat
4021 ggcgatgatc cgctgcaccg cgggatcgga cggatttgcg atggtgtcca actcagtcat
4081 ggtcgtccta ccggctgctg tgttcagtga cgcgattcct ggggtgtgac accctacgcg
4141 acgatggcgg atggctgccc tgaccggcaa tcaccaacgc aaggggaagt cgtcgctctc
4201 tggcaaagct ccccgctctt ccccgtccgg gacccgcgcg gtcgatcccc gcatatatga
4261 agtattcgcc ttgatcccca tcgaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
4321 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
4381 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
4441 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
4501 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
4561 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
4621 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
4681 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
4741 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
4801 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
4861 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
4921 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
4981 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
5041 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
5101 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
5161 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
5221 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
5281 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
5341 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttggt
5401 tcatgtgcag ctccatcagc aaaaggggat gataagttta tcaccaccga ctatttgcaa
5461 cagtgccgtt gatcgtgcta tgatcgactg atgtcatcag cggtggagtg caatgtcgtg
5521 caatacgaat ggcgaaaagc cgagctcatc ggtcagcttc tcaaccttgg ggttaccccc
5581 ggcggtgtgc tgctggtcca cagctccttc cgtagcgtcc ggcccctcga agatgggcca
5641 cttggactga tcgaggccct gcgtgctgcg ctgggtccgg gagggacgct cgtcatgccc
5701 tcgtggtcag gtctggacga cgagccgttc gatcctgcca cgtcgcccgt tacaccggac
5761 cttggagttg tctctgacac attctggcgc ctgccaaatg taaagcgcag cgcccatcca
5821 tttgcctttg cggcagcggg gccacaggca gagcagatca tctctgatcc attgcccctg
5881 ccacctcact cgcctgcaag cccggtcgcc cgtgtccatg aactcgatgg gcaggtactt
5941 ctcctcggcg tgggacacga tgccaacacg acgctgcatc ttgccgagtt gatggcaaag
6001 gttccctatg gggtgccgag acactgcacc attcttcagg atggcaagtt ggtacgcgtc
6061 gattatctcg agaatgacca ctgctgtgag cgctttgcct tggcggacag gtggctcaag
6121 gagaagagcc ttcagaagga aggtccagtc ggtcatgcct ttgctcggtt gatccgctcc
6181 cgcgacattg tggcgacagc cctgggtcaa ctgggccgag atccgttgat cttcctgcat
6241 ccgccagagg cgggatgcga agaatgcgat gccgctcgcc agtcgattgg ctgagctcat
6301 gagcggagaa cgagatgacg ttggaggggc aaggtcgcgc tgattgctgg ggcaacacgt
6361 ggagcggatc ggggattgtc tttcttcagc tcgctgatga tatgctgacg ctcaatgccg
6421 tttggcctcc gactaacgaa aatcccgcat ttggacggct gatccgattg gcacggcgga
6481 cggcgaatgg cggagcagac gctcgtccgg gggcaatgag atatgaaaaa gcctgaactc
6541 accgcgacgt atcgggccct ggccagctag ctagagtcga cctgcaggtc cccggggatc
6601 ggtcttgcct tgctcgtcgg tgatgtactt caccagctcc gcgaagtcgc tcttcttgat
6661 ggagcgcatg gggacgtgct tggcaatcac gcgcaccccc cggccgtttt agcggctaaa
6721 aaagtcatgg ctctgccctc gggcggacca cgcccatcat gaccttgcca agctcgtcct
6781 gcttctcttc gatcttcgcc agcagggcga ggatcgtggc atcaccgaac cgcgccgtgc
6841 gcgggtcgtc ggtgagccag agtttcagca ggccgcccag gcggcccagg tcgccattga
6901 tgcgggccag ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg tagccctggc
6961 cgacggccag caggtaggcc gacaggctca tgccggccgc cgccgccttt tcctcaatcg
7021 ctcttcgttc gtctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct
7081 tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc
7141 gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac
7201 acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc
7261 aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat
7321 ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa aagatccgtc
7381 gacctgcagg catgctaaac cgatacaatt aaaggctcct tttggagcct ttttttttgg
7441 aggcgaactc ccttatgcgg gtggcgcaga atgccaggac ccttgtcatt ccacgtcaat
7501 tcatgcgcct tttcacctcg tactgtcctg ccaaacacaa gcaacggagg tacg
//
