| 质粒名称: | pTW051d |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 116892 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | ColE1 |
| 载体出品公司: | -- |
| 载体抗性: | 大观霉素和链霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 6540 bp |
| 载体修饰: | -- |
| 载体类型: | -- |
| 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: | g3c |
| 物种来源: | Synthetic |
| 插入大小: | 1362 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pTW051d was a gift from David Liu (Addgene plasmid # 116892 ; http://n2t.net/addgene:116892 ; RRID:Addgene_116892) |
| 参考文献部分: | Continuous directed evolution of proteins with improved soluble expression. Wang T, Badran AH, Huang TP, Liu DR. Nat Chem Biol. 2018 Aug 20. pii: 10.1038/s41589-018-0121-5. doi: 10.1038/s41589-018-0121-5. 10.1038/s41589-018-0121-5 [pii] PubMed 30127387 |
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 pTW051d 6540 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION low stringency SE-PACE AP.
ACCESSION .
VERSION .
KEYWORDS pTW051d
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6540)
AUTHORS Wang T, Badran AH, Huang TP, Liu DR
TITLE Continuous directed evolution of proteins with improved soluble
expression.
JOURNAL Nat Chem Biol. 2018 Aug 20. pii: 10.1038/s41589-018-0121-5. doi:
10.1038/s41589-018-0121-5.
PUBMED 30127387
REFERENCE 2 (bases 1 to 6540)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6540
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(1..792)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
promoter complement(793..864)
/gene="bla"
/label=AmpR promoter
rep_origin 893..1481
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1382..1401
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1635..1652
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 1667..1807
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(1733..1752)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1893..1915
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(1909..2100)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
terminator 6120..6147
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 6239..6325
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 6478..6495
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
ORIGIN
1 ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg
61 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag
121 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt
181 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt
241 tcgctcatct ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc
301 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa
361 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc
421 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg
481 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc
541 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc
601 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg
661 tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc
721 gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac
781 cgcttccctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
841 catgagcgga tacatatttg aatgtattta gaaaaatagg ccaaataggc cgttgagatc
901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
961 tttgtttgcc ggatcaagag ctaccacctc tttttccgaa ggtaactggc ttcagcagag
1021 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
1081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
1141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
1201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
1261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
1321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
1381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
1441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
1501 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
1561 ctgattctgt ggataaccgt attacagcct ttgagtgagc tgataccgct cgccgcagcc
1621 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
1681 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
1741 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
1801 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtcttctccc
1861 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
1921 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
1981 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
2041 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
2101 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
2161 gaggatgctc acgatacggg ttactgatga tgaacaagag gacatccggt caaataaaac
2221 gaaaggctca gtcgaaagac tgggcctttc gttttgctga ggagacttag ggaccctact
2281 aatacgtctc actataggga gaaagacctg caggtgcagt aaaaaaggaa aaaaatgatc
2341 aaaattgcca cgcgtaaata tttaggcaaa cagaatgttt atgatatcgg tgtcgagcgc
2401 gatcataatt tcgcgctgaa aaacggcttt atcgccagca attgttttaa tgttgttcct
2461 ttctattctc actccgctga aactgttgaa agttgtttag caaaacccca tacagaaaat
2521 tcatttacta acgtctggaa agacgacaaa actttagatc gttacgctaa ctatgagggc
2581 tgtctgtgga atgctacagg cgttgtagtt tgtactggtg acgaaactca gtgttacggt
2641 acatgggttc ctattgggct tgctatccct gaaaatgagg gtggtggctc tgagggtggc
2701 ggttctgagg gtggcggttc tgagggtggc ggtactaaac ctcctgagta cggtgataca
2761 cctattccgg gctatactta tatcaaccct ctcgacggca cttatccgcc tggtactgag
2821 caaaaccccg ctaatcctaa tccttctctt gaggagtctc agcctcttaa tactttcatg
2881 tttcagaata ataggttccg aaataggcag ggggcattaa ctgtttatac gggcactgtt
2941 actcaaggca ctgaccccgt taaaacttat taccagtaca ctcctgtatc atcaaaagcc
3001 atgtatgacg cttactggaa cggtaaattc agagactgcg ctttccattc tggctttaat
3061 gaggatccat tcgtttgtga atatcaaggc caatcgtctg acctgcctca acctcctgtc
3121 aatgctggcg gcggctctgg tggtggttct ggtggcggct ctgagggtgg tggctctgag
3181 ggtggcggtt ctgagggtgg cggctctgag ggaggcggtt ccggtggtgg ctctggttcc
3241 ggtgattttg attatgaaaa gatggcaaac gctaataagg gggctatgac cgaaaatgcc
3301 gatgaaaacg cgctacagtc tgacgctaaa ggcaaacttg attctgtcgc tactgattac
3361 ggtgctgcta tcgatggttt cattggtgac gtttccggcc ttgctaatgg taatggtgct
3421 actggtgatt ttgctggctc taattcccaa atggctcaag tcggtgacgg tgataattca
3481 cctttaatga ataatttccg tcaatattta ccttccctcc ctcaatcggt tgaatgtcgc
3541 ccttttgtct ttggcgctgg taaaccttac gagttcagta tcgactgcga taagatcaac
3601 ctgttccgcg gtgtctttgc gtttctttta tatgttgcca cctttatgta tgtattttct
3661 acgtttgcta acatactgcg taataaggag tcttaaacgg agccaatgta cgcaaaaaac
3721 cccgcttcag cggggttttt tcgccaaaaa aaaccccgcc ctgtcagggg cggggttttt
3781 ttttagggcc ctactgactg ttacgcgaac gcgaagtccg actctaagat gtcacggagg
3841 ttcaagttac ctttagccgg aagtgctggc attttgtcca attgagactc gtgcaactgg
3901 tcagcgaact ggtcgtagaa atcagccagt acatcacaag actcatatgt gtcaaccata
3961 gtttcgcgca ctgctttgaa caggttcgca gcgtcagccg gaatggtacc gaaggagtcg
4021 tgaatcagtg caaaagattc gattccgtac ttctcgtgtg cccacactac agtcttacga
4081 aggtggctac cgtcttggct gtgtacaaag ttaggagcga taccagactc ctgtttgtgt
4141 gcatcaatct cgctatcttt gttggtgtta atggtaggct gtaagcggaa ctgaccgagg
4201 aacatcaggt tcaagcgcgt ctgaataggc ttcttgtatt cctgccacac agggaaacca
4261 tcaggagtta cccaatgcac agcgcaacgc ttgcgaagaa tctctccagt cttcttatct
4321 ttgacctcag cagccagcag cttagcagca gacttaagcc agttcattgc ttcaaccgca
4381 gctaccaccg tcacgctcac agattcccaa atcagcttag ccatgtatcc agcagcctga
4441 ttcggctgag tgaacatcag acccttgccg gaatcaatag ctggctgaat ggtatcttcc
4501 agcacacttt gacggaagcc gaactctttg gacccgtaag ccagcgtcat gactgaactc
4561 ttagtcacac tgcgagtaac accgtaagcc agccattgac cagccagtgc cttagtgccc
4621 agcttgactt tctcagagat ttcaccagtg ttctcatcgg tcacggtaac tacttcgtta
4681 tcggtcccat tgattgcgtc tgcttgtaga atctcgttga ctttcttagc aacaatcccg
4741 tagatgtcct gaacggtttc actaggaagc aagttaaccg cgcgaccacc tacctcatct
4801 cggagcatcg cggagaagtg ctggatgcca gagcaagacc cgtcaaacgc cagcggaagg
4861 gagcagttat agctcaggcc gtggtgctgt accccagcgt actcaaagca gaacgcaagg
4921 aagcagaacg gagaatcttg ctcagcccac caagtgttct ccagtggaga cttagcgcaa
4981 gccatgatgt tctcgtggtt ttcctcaatg aacttgatgc gctcagggaa cggaacctta
5041 tcgacacccg cacagtttgc accgtggatt ttcagccagt agtaaccttc cttaccgatt
5101 ggtttacctt tcgccagcgt aagcagtcct ttggtcatat cgttaccttg cgggttgaac
5161 attgacacag cgtaaacacg accgcgccag tccatgttgt aagggaacca gatggcctta
5221 tggttagcaa acttattggc ttgctcaagc atgaactcaa ggctgatacg gcgagacttg
5281 cgagccttgt ccttgcggta cacagcagcg gcagcacgtt tccacgcggt gagagcctca
5341 ggattcatgt cgatgtcttc cggtttcatc gggagttctt cacgctcaat cgcagggatg
5401 tcctcgaccg gacaatgctt ccacttggtg attacgttgg cgaccgctag gactttcttg
5461 ttgattttcc atgcggtgtt ttgcgcaatg ttaatcgctt tgtacacctc aggcatgtaa
5521 acgtcttcgt agcgcatcag tgctttctta ctgtgagtac gcaccagcgc cagaggacga
5581 cgaccgttag cccaatagcc accaccagta atgccagtcc acggcttagg aggaactacg
5641 caaggttgga acatcggaga gatgccagcc agcgcacctg cacgggttgc gatagcctca
5701 gcgtattcag gtgcgagttc gatagtctca gagtcttgac ctactacgcc agcattttgg
5761 cggtgtaagc taaccattcc ggttgactca atgagcatct cgatgcagcg tactcctaca
5821 tgaatagagt cttccttatg ccacgaagac cacgcctcgc caccgagtag acccttagag
5881 agcatgtcag cctcgacaac ttgcataaat gctttcattt ttttttcctc cttactctag
5941 taaaagttaa acaaaattat ttgtagaggg aaaccgttgt ggtctccctg aatatagata
6001 ccgagcctta tgcatgcccg taaagttatc cagcaaccac tcatagacct agggcagcag
6061 atagggacga cgtggtgtta gctgtgctct agaatcctca gcgagtttgt agaaacgcaa
6121 aaaggccatc cgtcaggatg gccttctgct taatttgatg cctggcagtt tatggcgggc
6181 gtcctgcccg ccaccctccg ggccgttgct tcgcaacgtt caaatccgct cccggcggat
6241 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
6301 tcgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc atggggagac
6361 cccacactac catcggcgct acggcgtttc acttctgagt tcggcatggg gtcaggtggg
6421 accaccgcgc tactgccgcc aggcaaattc tgttttatca gaccgcttct gcgttctgat
6481 ttaatctgta tcaggctgaa aatcttctct catccgccaa aacagccatg cccatggcgt
//
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