| 质粒名称: | pSL1521 (pSPIN, pSC101* backbone) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 160729 |
| 存储实验室: | Samuel H. Sternberg |
|
载体骨架基本信息 |
|
| 载体名称: | pSC101* |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 12724bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | Spectinomycin, 50 μg/mL |
| 生长温度: | 30℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | VchCAST proteins, crRNA, and donor DNA |
| 物种来源: | V. cholerae |
| 启动子: | J23119 |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | CGCAGATCAGTTGGAAGAAT |
| 3’测序引物 | GCTAGTTATTGCTCAGCGG |
|
文献引用方法 |
|
| 材料和方法部分: | pSL1521 (pSPIN, pSC101* backbone) was a gift from Samuel H. Sternberg (Addgene plasmid # 160729 ; http://n2t.net/addgene:160729 ; RRID:Addgene_160729) |
| 参考文献部分: | CRISPR RNA-guided integrases for high-efficiency, multiplexed bacterial genome engineering. Vo PLH, Ronda C, Klompe SE, Chen EE, Acree C, Wang HH, Sternberg SH. Nat Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi: 10.1038/s41587-020-00745-y. 10.1038/s41587-020-00745-y PubMed 33230293 |
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pSL1521(pSPIN,pSC101*backbone)载体质粒基本信息
pSL1521(pSPIN,pSC101*backbone)质粒图谱载体图谱和pSL1521(pSPIN,pSC101*backbone)载体序列质粒序列多克隆位点信息
pSL1521(pSPIN,pSC101*backbone)质粒载体简介
pSL1521(pSPIN,pSC101*backbone)质粒序列载体序列
LOCUS pSL1521 (pSPIN, pSC101* backbone) 12724 bp DNA circular SYN 25-AUG-2024
DEFINITION Single-plasmid V. cholerae CAST system. Encodes all proteins, crRNA,
donor DNA; non-targeting crRNA has BsaI sites for cloning.
Temperature-sensitive pSC101* backbone can be cured by 37 (o)C
incubation..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12724)
AUTHORS Vo PLH, Ronda C, Klompe SE, Chen EE, Acree C, Wang HH, Sternberg SH
TITLE CRISPR RNA-guided integrases for high-efficiency, multiplexed
bacterial genome engineering.
JOURNAL Nat Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi:
10.1038/s41587-020-00745-y.
PUBMED 33230293
REFERENCE 2 (bases 1 to 12724)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi:
10.1038/s41587-020-00745-y."
FEATURES Location/Qualifiers
source 1..12724
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 241..332
/gene="bla"
/label=AmpR promoter
CDS 333..1124
/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"
primer_bind 1286..1303
/label=L4440
/note="L4440 vector, forward primer"
promoter 1310..1344
/label=J23119 promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119)"
CDS 8326..8337
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(9772..9790)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 9786..9833
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS 9988..10647
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(10054..10073)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
primer_bind complement(10847..10864)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(10847..10863)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10856..10878)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 11470..11692
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS 11740..12690
/codon_start=1
/gene="rep101 encoding an A56V mutation"
/product="temperature-sensitive version of the RepA protein
needed for replication with the pSC101 origin (Armstrong et
al., 1984)"
/label=Rep101(Ts)
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
1 tcaagactaa caaacaaaag tagaacaact gttcaccgtt acatatcaaa gggaaaactg
61 tccataccca tggattcttc gtctgtttct actggtattg gcacaaacct gatatcgatg
121 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
181 attcaaatat gtatccgctc atgagacaat aaccctggta gcgaccgagt gagctagcta
241 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
301 aaatgcttca ataatattga aaaaggaaga gtatgaggga agcggtgatc gccgaagtat
361 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
421 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
481 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
541 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
601 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
661 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
721 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
781 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
841 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
901 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
961 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
1021 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
1081 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataatgtcct gtcagcagac
1141 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
1201 taggtgaaga tccgcctgca tggataaccg tattaccgcc tttgagtgag ctgataccgc
1261 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagctt tgacagctag
1321 ctcagtccta ggtataatac tagcgtcgac gtggagatat accatgggtg aactgccgag
1381 taggtagctg ataacgagac ctctgtcttg tcagctaggg tggtctcgtg aactgccgag
1441 taggtagctg ataacggatc cgaattcgag cgaaggagat atacatatgt ttttgcaaag
1501 acctaaacct tacagcgatg aaagtttaga aagtttcttt atccgagtgg ctaacaaaaa
1561 tggctacggt gatgtccatc gcttcctaga agccactaaa cgattccttc aagacattga
1621 ccataatggc tatcaaacct ttccgactga tataactcgg ataaacccat actcagctaa
1681 aaacagttcc agcgcacgaa ctgcgtcatt cctgaaactt gcacaattga catttaatga
1741 accgccagag ctacttgggt tggcaattaa cagaacaaac atgaaatact cgccgtcaac
1801 tagcgcggtt gttagaggtg cagaagtctt tcctcgcagt ttactacgga cgcactccat
1861 cccctgctgt cctttgtgtc tgcgagaaaa tggctacgcc tcctaccttt ggcactttca
1921 ggggtacgaa tactgccaca gccataacgt acctttaatt accacttgta gctgtggtaa
1981 ggagtttgac taccgagtat ctgggttaaa gggcatttgc tgcaaatgca aggagcctat
2041 caccttaacc agcagggaga acggtcatga ggcagcgtgt actgtttcaa actggcttgc
2101 tggccatgaa tctaaacctc tgccaaatct tcctaaaagc taccgatggg gtttagttca
2161 ttggtggatg ggtattaaag atagcgagtt cgatcacttt tcgttcgttc aatttttctc
2221 aaactggcca aggtcattcc actcgataat cgaagatgaa gtagagttca accttgagca
2281 tgctgttgtc agcacgtctg aattacgact aaaagatctt cttggtcgat tgtttttcgg
2341 ttcaattcgg ttacctgagc ggaatcttca acacaatatc atccttggtg agcttctctg
2401 ctatttagaa aatcgtttat ggcaagacaa gggattaatc gccaacctca aaatgaacgc
2461 gttagaggcg actgtaatgt taaattgtag cctcgatcag attgcatcaa tggttgaaca
2521 acgcatcttg aagccaaatc gaaaaagcaa gcccaacagc cctcttgatg ttaccgatta
2581 tctatttcat ttcggcgata ttttctgtct ttggttagct gagttccaaa gcgatgagtt
2641 taaccgttcg ttttatgtgt cgaggtggta aatgcaaact ctgaaagaac taatcgcatc
2701 caatcctgac gacttaacaa ctgaacttaa gagagcattt cgtccactca caccgcatat
2761 tgcaattgat ggtaatgaac ttgacgcact gacgatatta gtcaatttaa ccgataagac
2821 tgatgatcag aaagacctgc tcgatcgagc caaatgcaag caaaaacttc gagatgaaaa
2881 atggtgggct agctgcataa attgcgttaa ttacagacaa agccataacc caaaattccc
2941 ggatatacgt tctgaaggcg tgattcgaac ccaagccctg ggtgaattac cgagtttcct
3001 actctcgtcc tcaaaaatcc caccatacca ttggtcatat agccatgatt caaaatacgt
3061 caacaaaagc gcattcctca ccaatgagtt ttgttgggat ggtgagatct catgtttggg
3121 tgagcttctt aaagatgcag atcacccact ttggaatact ttgaaaaagt taggttgttc
3181 tcaaaaaact tgcaaagcaa tggcaaaaca actagctgat attactctca cgactatcaa
3241 tgtcacactc gcaccaaatt acctgactca aatatctctc cccgatagtg atacatccta
3301 catttcactc tctcctgttg catcgctatc gatgcaaagc cactttcatc agaggctcca
3361 agatgaaaat cggcatagcg cgataacgcg gtttagccga actaccaaca tgggggtcac
3421 tgcgatgaca tgtggcggcg catttagaat gttaaagtct ggcgctaagt tttccagccc
3481 ccctcatcac cgattaaaca gtaaacgaag ttggttgacg tcagagcatg ttcagtcatt
3541 aaaacagtac cagcgcctca acaaaagcct catacctgaa aactctcgga ttgcactccg
3601 tagaaaatac aaaatcgagc ttcaaaatat ggtcagatct tggtttgcaa tgcaagacca
3661 tacccttgat tcgaacatac ttatccaaca tttgaatcat gacctatctt acttaggagc
3721 cacaaaacgt tttgcatacg atccagcgat gaccaagctc tttactgagc ttttgaaacg
3781 agagttatca aattcaatca ataatggtga gcaacacact aatggatcgt ttttagtcct
3841 accgaatatc agggtttgtg gcgcaacagc tttaagctcc ccggtaacgg tggggattcc
3901 atcacttaca gctttctttg gcttcgttca cgcatttgaa cggaatataa atcgcaccac
3961 ctcatcgttt cgtgttgaat cctttgcgat atgcgtccat caactacatg tcgagaagcg
4021 aggtttgaca gcagagtttg tggaaaaagg cgacgggact atatccgctc ccgcgacccg
4081 ggatgactgg cagtgtgatg tcgtatttag ccttattttg aacaccaact ttgctcaaca
4141 tattgaccaa gatacgttag ttacatcact accaaagcga ttggctcggg gttcagcaaa
4201 aattgcgatt gatgacttta aacatatcaa ctcattctcg acattagaaa cagcgatcga
4261 atctctgcca atagaagctg gtaggtggtt atcactttac gcacagtcaa acaataatct
4321 aagtgatcta ttagcagcca tgacagagga ccatcagctc atggcaagct gcgtcggtta
4381 ccacttgtta gaagagccca aagataaacc aaactccctc agaggttaca aacacgctat
4441 cgccgagtgc atcattggac tcattaactc aatcaccttt agctcagaga ctgatcccaa
4501 cacaatcttt tggtcgctaa agaactatca aaactaccta gtggtacagc caaggagtat
4561 caacgatgaa actaccgaca aatctagcct atgagcgctc tatcgaccca tcagatgtct
4621 gtttttttgt cgtctggccc gatgatagaa aaacaccttt aacctacaat tctcgtactc
4681 tgctcgggca aatggaagcg gcatcattag cctatgatgt ctcaggtcaa ccaataaaaa
4741 gtgccaccgc tgaggcgtta gctcaaggga accctcatca agttgatttc tgccacgttc
4801 catacggtgc gagtcatatt gaatgcagtt tctccgtctc gttttcttct gaactacgtc
4861 aaccatataa gtgtaactca agcaaagtta aacaaacgct agtgcaatta gtcgagctct
4921 acgaaacgaa aatcggctgg actgagctag caacccgata tttgatgaac atttgcaacg
4981 gtaaatggct gtggaaaaat acccgtaaag cctattgctg gaacattgta cttacacctt
5041 ggccttggaa cggggaaaag gttggatttg aagatattcg tactaactac acctcacggc
5101 aagactttaa aaataataaa aattggtctg ctatagttga aatgatcaaa accgcatttt
5161 ctagtactga tgggctggcg atatttgaag tcagggccac cttgcacttg ccaacgaatg
5221 ctatggtgcg gccaagccaa gttttcacag aaaaagaaag tggcagtaaa agtaaatcta
5281 aaactcaaaa cagtcgagtt tttcagagta caactattga tggtgaacga tcgccaatac
5341 taggggcctt taaaacggga gcagctattg caaccattga cgactggtat cctgaagcca
5401 ctgagccact aagggtcgga cggtttgggg ttcatcgcga agatgtcact tgctaccgtc
5461 atccgtctac cggaaaagat tttttctcga tattacaaca agcagagcac tatattgaag
5521 tgttgagcgc caacaaaact cccgctcaag aaactatcaa cgacatgcac tttttaatgg
5581 ctaacctgat taagggtggg atgttccagc ataaaggaga ctgactgtga aatggtatta
5641 taagacaatc acctttctgc cagagttgtg caacaacgag tcactggctg caaagtgtct
5701 ccgcgttctg catggattta actatcagta tgagacacga aatatcggcg tttcatttcc
5761 gctttggtgt gatgcaacgg ttggaaaaaa gatttcattt gtcagcaaga acaagataga
5821 actcgactta ctacttaaac aacactattt cgtccaaatg gaacaacttc aatactttca
5881 tatatccaac actgttctcg tcccagaaga ttgtacatac gtttccttta gacgctgtca
5941 atctatagat aagctcacag cagcagggct ggcaaggaaa atcagacgcc tggagaaacg
6001 tgctctatct agaggcgagc aatttgaccc atcatctttt gctcaaaaag agcatactgc
6061 aatagcgcac taccactcac ttggggagtc cagcaaacag acgaaccgca actttcgact
6121 caatatcagg atgctctcgg agcaaccccg tgaggggaac tcgattttta gtagctatgg
6181 cttatcaaat tcagaaaact cgtttcagcc tgtaccctta atctgacctt taataaggag
6241 atataccatg gcgacaagtt tacctacgcc ctcagcaatt acgacttcgg cgttagagta
6301 tgcattccat actcccgctc gcaatctaac gaaatctcgc ggaaaaaaca ttcatcgtta
6361 tgtcagtgta aagatgagta agaggattac ggtagaatct actctagagt gtgatgcctg
6421 ctatcacttt gattttgagc caagtattgt tcgcttttgc gctcaaccga ttcgattttt
6481 atattatctc aatggtcagt ctcactccta tgttcctgac tttctagttc aatttgatac
6541 caacgagttt gttctatatg aagtaaagtc agcttatgct aagaacaaac ctgattttga
6601 tgttgaatgg gaggcgaaag taaaagcagc aactgaacta gggctagaat tggagcttgt
6661 tgaagagagt gatattaggg atacggttgt attaaataat cttaagcgca tgcatcgtta
6721 tgcttcgaaa gatgagctga ataacgtaca taactctctc ttaaaaataa taaagtacaa
6781 tggcgcccaa tctgcaagat gcttgggaga acagttgggt ttaaaaggcc gaactgtttt
6841 accaattttg tgcgatttgc tgtcaaggtg tttactcgat acacgtttgg ataagcctct
6901 atctcttgaa tctcgatttg agttggccag ttatggctaa gaaagggttc tcaagtttcc
6961 atagaaaagc agtctcttcc caagatacgc ttgaatcgat agagcttgtc tctagcgcta
7021 attgtctaga aagtgttacg tatcaagata tatcagcatt tcccgaaaca attgcggtag
7081 agattaattt ccgattaagc attcttcgct ttttagcgcg aaagtgcgaa accattgtgg
7141 ccaaatcaat tgaaccacat cgtgtagagc tacagcaaaa ctatagtaga aaaataccca
7201 gtgcaataac gatatatcga tggtggcttg cttttcgaaa atcagactac aaccccatta
7261 gcttagcacc taatatcaag gatagaggta atagagagac aaaagtgtca acagttgttg
7321 attctattat ggaacaggca gttgaaagag ttatatctgg acgaaaagtc aatgttagct
7381 ctgcatataa acgtgttcga cgaaaagttc gtcaatacaa tctaactcat ggaacgaaat
7441 acacgtatcc taagtacgaa tctgtaagaa agcgagtaaa aaagaaaacc ccatttgagt
7501 tattagccgc agggaaaggg gagagagtag ctaagagaga gtttcgccga atgggaaaaa
7561 agatcctcac gtctagcgtg ctagagaggg ttgaaataga tcacactgtc gttgaccttt
7621 ttgcagtaca cgaagagtat cgaatcccat tgggccgacc ttggcttact caattggttg
7681 attgttacag taaagctgta atcggttttt atttaggttt cgagcctcct agctatgtgt
7741 cggtttccct tgcacttaag aatgcaatac aacgcaaaga tgacttaatc tcctcgtatg
7801 aatcgatcga gaatgaatgg ctatgttatg gcatcccaga cctactcgta actgataatg
7861 gtaaagagtt tttgtcgaaa gcatttgatc aagcatgtga atcactattg atcaatgtgc
7921 atcaaaataa agttgagacg cccgacaaca aacctcatgt tgaacgtaac tacgggacta
7981 ttaatacttc tctgttagac gatttacctg ggaaatcctt cagccagtac cttcaaagag
8041 aagggtacga ctctgtggga gaagctaccc ttacactcaa tgagattaga gaaatttact
8101 taatttggtt ggtggatatt tatcataaaa aacccaatca gagaggcact aattgtccta
8161 atgttgcgtg gaaaaagggt tgtcaagaat gggaaccaga ggagttctct ggttctaaag
8221 acgaattaga ctttaaattt gctattgttg attacaaaca acttactaaa gtagggataa
8281 ctgtctacaa agaactgagt tatagcaatg accgtttagc tgaatataga gggaagaaag
8341 gaaaccataa agttcagttc aagtataacc ctgagtgtat ggcagttatt tgggtgttgg
8401 atgaggatat gaatgagtac tttacagtta atgcgattga ctacgaatat gcaagtagag
8461 tatcactttg gcaacataaa tataacatga aatatcaagc agaactaaat tcagcagaat
8521 atgatgagga caaggaaatt gatgcagaaa taaaaattga agaaatcgca gatcgttcaa
8581 ttgttaagac taacaaaatc agagctcgga ggcgtggcgc taggcatcaa gagaatagcg
8641 caagggctaa gtcaatcagt aatgcgaacc cggcctcgat acaaaaacat gaagatgaaa
8701 tcgttagtgc agataatgac gattgggata ttgattatgt ctgagaaatc gtcaaatgag
8761 tgaaacgcgt gaggctcgta tatcaagagc taaaagggca tttgtatcca caccgagcgt
8821 taggaaaatc ttgagttaca tggatagatg tagagatcta tcagacctag agtctgagcc
8881 tacatgcatg atggtctatg gtgcttcggg tgtaggtaaa acgaccgtca tcaagaaata
8941 cttaaatcaa aacagaagag agtccgaagc cgggggcgat ataataccgg ttttgcatat
9001 tgagttgcca gacaatgcga agccagtaga tgcagcaagg gaattgctgg ttgaaatggg
9061 tgacccgcta gcactttatg aaactgactt agctagattg acgaaaagac tgactgaatt
9121 aatccctgcg gtcggcgtga agctgattat tatcgatgag ttccaacatt tggtggaaga
9181 aaggtcaaat cgggttctta cccaagtagg taattggcta aaaatgatac ttaacaaaac
9241 gaaatgtcca attgttatat ttggtatgcc atactcaaaa gttgtactgc aagcaaactc
9301 gcaacttcac gggcgatttt ccattcaggt tgaactgcgc ccctttagct accagggagg
9361 tagaggtgta tttaaaactt ttttggaata ccttgataaa gccctacctt ttgaaaaaca
9421 ggctggctta gccaacgaaa gtttgcagaa aaaattgtat gcattctctc agggaaacat
9481 gcgttcgttg agaaacctta tttatcaagc atctatcgaa gcaattgata atcagcatga
9541 gacgataacc gaagaagatt tcgtttttgc atcgaagttg acatcgggcg ataaacccaa
9601 ctcatggaaa aatccttttg aggagggtgt tgaggtaaca gaagatatgt tacgaccgcc
9661 accaaaagat attggttggg aggactattt gagacattca accccgagag tgagtaaacc
9721 aggtagaaat aaaaactttt tcgaataacc taggctgctg cgctgctgcc accgctgagc
9781 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac
9841 ctcaggcatt tgttgttgat acaaccataa aatgataatt acacccataa attgataatt
9901 atcacaccca taaattgata ttgcctcttc atggtctaaa cttcagtaag tttacgacat
9961 tttcctcgag gtcatttccg gggatccatg gagaaaaaaa tcactggata taccaccgtt
10021 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
10081 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
10141 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
10201 gagttccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
10261 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
10321 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
10381 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
10441 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
10501 actatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
10561 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
10621 tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttctaga
10681 gtccttactg cagtagtttt gctgaaatac tcgattcaca aaaatatcaa cttatggttg
10741 ttttgtgaga tatcaatata tggttgtttt gtggttaagt tgctgattat aaataattat
10801 taaatatcac tttatggttg catcaacagg taccgagctc gaattcactg gccgtcgttt
10861 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gccagttcac
10921 cgacaaacga aagctgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
10981 tgagcgtcag accccgttga tgataccgct gccttactgg gtgcattagc cagtctgaat
11041 gacctgtcac gggataatcc gaagtggtca gactggaaaa tcagagggca ggaactgctg
11101 aacagcaaaa agtcagatag caccacatag cagacccgcc ataaaacgcc ctgagaagcc
11161 cgtgacgggc ttttcttgta ttatgggtag tttccttgca tgaatccata aaaggcgcct
11221 gtagtgccat ttacccccat tcactgccag agccgtgagc gcagcgaact gaatgtcacg
11281 aaaaagacag cgactcaggt gcctgatggt cggagacaaa aggaatattc agcgatttgc
11341 ccgattgcgg ccgcaaccga gcttgcgagg gtgctactta agcctttagg gttttaaggt
11401 ctgttttgta gaggagcaaa cagcgtttgc gacatccttt tgtaatactg cggaactgac
11461 taaagtagtg agttatacac agggctggga tctattcttt ttatcttttt ttattctttc
11521 tttattctat aaattataac cacttgaata taaacaaaaa aaacacacaa aggtctagcg
11581 gaatttacag agggtctagc agaatttaca agttttccag caaaggtcta gcagaattta
11641 cagataccca caactcaaag gaaaaggact agtaattatc attgactagc ccatctcaat
11701 tggtatagtg attaaaatca cctagaccaa ttgagatgta tgtctgaatt agttgttttc
11761 aaagcaaatg aactagcgat tagtcgctat gacttaacgg agcatgaaac caagctaatt
11821 ttatgctgtg tggcactact caaccccacg attgaaaacc ctacaaggaa agaacggacg
11881 gtatcgttca cttataacca atacgttcag atgatgaaca tcagtaggga aaatgcttat
11941 ggtgtattag ctaaagcaac cagagagctg atgacgagaa ctgtggaaat caggaatcct
12001 ttggttaaag gctttgagat tttccagtgg acaaactatg ccaagttctc aagcgaaaaa
12061 ttagaattag tttttagtga agagatattg ccttatcttt tccagttaaa aaaattcata
12121 aaatataatc tggaacatgt taagtctttt gaaaacaaat actctatgag gatttatgag
12181 tggttattaa aagaactaac acaaaagaaa actcacaagg caaatataga gattagcctt
12241 gatgaattta agttcatgtt aatgcttgaa aataactacc atgagtttaa aaggcttaac
12301 caatgggttt tgaaaccaat aagtaaagat ttaaacactt acagcaatat gaaattggtg
12361 gttgataagc gaggccgccc gactgatacg ttgattttcc aagttgaact agatagacaa
12421 atggatctcg taaccgaact tgagaacaac cagataaaaa tgaatggtga caaaatacca
12481 acaaccatta catcagattc ctacctacat aacggactaa gaaaaacact acacgatgct
12541 ttaactgcaa aaattcagct caccagtttt gaggcaaaat ttttgagtga catgcaaagt
12601 aagtatgatc tcaatggttc gttctcatgg ctcacgcaaa aacaacgaac cacactagag
12661 aacatactgg ctaaatacgg aaggatctga ggttcttatg gctcttgtat ctatcagtga
12721 agca
//
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pSL1521(pSPIN,pSC101*backbone)质粒载体应用举例
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