企业认证
-20℃低温保存
三年
pCas
20
泽叶生物
| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pCas |
| 质粒类型: | CRISPR/Cas9载体；基因敲除载体;原核载体 |
| 高拷贝/低拷贝: | 低拷贝 |
| 克隆方法: | 限制性内切酶，多克隆位点 |
| 启动子: | native cas9 promoter |
| 载体大小: | 12545 bp |
| 5' 测序引物及序列: | pENTR1A-3 (GTAACATCAGAGATTTTGAGACAC) |
| 3' 测序引物及序列: | AraC-F-New (GATGTAGCCGTCAAGTTGTCA) |
| 载体标签: | NLS、His8 (C ter) |
| 载体抗性: | 卡那霉素(Kanamycin) |
| 筛选标记: | -- |
| 克隆菌株: | Stbl3 |
| 宿主细胞（系）: | 细菌 |
| 备注: | Must be grown at 30C! Vector has temperature-sensitive replication (RepA101ts) |
| 产品目录号: | #62225 |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 诱导型 |
| 病毒/非病毒: | 非病毒 |
LOCUS pCas 12545 bp DNA SYN
DEFINITION pCas
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..12545
/organism="pCas"
/mol_type="other DNA"
misc_feature complement(404..1495)
/label="lacI"
CDS complement(404..1363)
/label="ORF frame 2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS complement(1698..2648)
/label="ORF frame 1"
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRK
ERTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAK
FSSEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHK
ANIEISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTL
IFQVELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTS
FEAKFLSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI*"
gene complement(3572..4387)
/label="KanR2"
/gene="KanR2"
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRK
ERTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAK
FSSEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHK
ANIEISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTL
IFQVELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTS
FEAKFLSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI*"
CDS complement(3572..4387)
/label="ORF frame 2"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
CDS 4917..9023
/label="ORF frame 3"
/translation="MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIK
KNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLE
ESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAH
MIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSK
SRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLD
NLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTL
LKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVK
LNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPY
YVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVL
PKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKE
DYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLT
LFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDF
LKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQT
VKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKE
HPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKV
LTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKA
GFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF
YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIG
KATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS
MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVV
AKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLF
ELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQH
KHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA
PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD*"
misc_feature complement(9153..10031)
/label="AraC"
/translation="MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIK
KNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLE
ESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAH
MIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSK
SRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLD
NLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTL
LKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVK
LNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPY
YVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVL
PKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKE
DYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLT
LFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDF
LKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQT
VKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKE
HPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKV
LTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKA
GFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF
YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIG
KATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS
MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVV
AKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLF
ELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQH
KHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA
PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD*"
CDS complement(9153..10082)
/label="ORF frame 1"
/translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIE
ANGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRH
PEAREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQG
EGRYSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVA
QHVCLSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQL
YFSRVFKKCTGASPSEFRAGCEEKVNDVAVKLS*"
misc_feature 10061..10076
/label="araO2"
/translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIE
ANGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRH
PEAREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQG
EGRYSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVA
QHVCLSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQL
YFSRVFKKCTGASPSEFRAGCEEKVNDVAVKLS*"
misc_feature 10218..10229
/label="araO1"
/translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIE
ANGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRH
PEAREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQG
EGRYSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVA
QHVCLSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQL
YFSRVFKKCTGASPSEFRAGCEEKVNDVAVKLS*"
misc_feature 10260..10273
/label="CAP_BS"
/translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIE
ANGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRH
PEAREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQG
EGRYSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVA
QHVCLSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQL
YFSRVFKKCTGASPSEFRAGCEEKVNDVAVKLS*"
misc_feature 10265..10284
/label="pBAD_fwd_primer"
/translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIE
ANGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRH
PEAREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQG
EGRYSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVA
QHVCLSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQL
YFSRVFKKCTGASPSEFRAGCEEKVNDVAVKLS*"
misc_feature 10270..10308
/label="AraI1I2"
/translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIE
ANGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRH
PEAREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQG
EGRYSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVA
QHVCLSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQL
YFSRVFKKCTGASPSEFRAGCEEKVNDVAVKLS*"
promoter 10305..10333
/label="ARA_promoter"
/translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIE
ANGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRH
PEAREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQG
EGRYSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVA
QHVCLSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQL
YFSRVFKKCTGASPSEFRAGCEEKVNDVAVKLS*"
CDS 10808..11593
/label="ORF frame 2"
/translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQF
IALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNE
SCTCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCA
RLAFGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDD
DLLPLCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA*"
CDS 11590..12270
/label="ORF frame 1"
/translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKP
RSGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTE
SPIIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQY
SMWVTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIG
FVFGEQWR*"
ORIGIN
1 gatctcaaaa aaagcaccga ctcggtgcca ctttttcaag ttgataacgg actagcctta
61 ttttaacttg ctatttctag ctctaaaacc tggtaacagg attagcagat gtgtgaaatt
121 gttatccgct cacaattcca cacattatac gagccggatg attaattgtc aacagctcat
181 ttcagaatat ttgccagaac cgttatgatg tcggcgcaaa aaacattatc cagaacggga
241 gtgcgccttg agcgacacga attatgcagt gatttacgac ctgcacagcc ataccacagc
301 ttccgatggc tgcctgacgc cagaagcatt ggtgcaccgt gcagtcgatg ataagctgtc
361 aaaccagatc aattcgcgct aactcacatt aattgcgttg cgctcactgc ccgctttcca
421 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
481 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
541 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
601 gcaggcgaaa atcctgtttg atggtggttg acggcgggat ataacatgag ctgtcttcgg
661 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
721 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
781 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
841 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
901 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
961 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
1021 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
1081 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
1141 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
1201 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
1261 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
1321 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
1381 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
1441 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
1501 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac cattcgatgg
1561 tgtcaacgta aatgcatgcc gcttcgcctt ccatgggtat ggacagtttt ccctttgata
1621 tgtaacggtg aacagttgtt ctacttttgt ttgttagtct tgatgcttca ctgatagata
1681 caagagccat aagaacctca gatccttccg tatttagcca gtatgttctc tagtgtggtt
1741 cgttgttttt gcgtgagcca tgagaacgaa ccattgagat catacttact ttgcatgtca
1801 ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg cagttaaagc atcgtgtagt
1861 gtttttctta gtccgttacg taggtaggaa tctgatgtaa tggttgttgg tattttgtca
1921 ccattcattt ttatctggtt gttctcaagt tcggttacga gatccatttg tctatctagt
1981 tcaacttgga aaatcaacgt atcagtcggg cggcctcgct tatcaaccac caatttcata
2041 ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa cccattggtt aagcctttta
2101 aactcatggt agttattttc aagcattaac atgaacttaa attcatcaag gctaatctct
2161 atatttgcct tgtgagtttt cttttgtgtt agttctttta ataaccactc ataaatcctc
2221 atagagtatt tgttttcaaa agacttaaca tgttccagat tatattttat gaattttttt
2281 aactggaaaa gataaggcaa tatctcttca ctaaaaacta attctaattt ttcgcttgag
2341 aacttggcat agtttgtcca ctggaaaatc tcaaagcctt taaccaaagg attcctgatt
2401 tccacagttc tcgtcatcag ctctctggtt gctttagcta atacaccata agcattttcc
2461 ctactgatgt tcatcatctg agcgtattgg ttataagtga acgataccgt ccgttctttc
2521 cttgtagggt tttcaatcgt ggggttgagt agtgccacac agcataaaat tagcttggtt
2581 tcatgctccg ttaagtcata gcgactaatc gctagttcat ttgctttgaa aacaactaat
2641 tcagacatac atctcaattg gtctaggtga ttttaatcac tataccaatt gagatgggct
2701 agtcaatgat aattactagt ccttttcctt tgagttgtgg gtatctgtaa attctgctag
2761 acctttgctg gaaaacttgt aaattctgct agaccctctg taaattccgc tagacctttg
2821 tgtgtttttt ttgtttatat tcaagtggtt ataatttata gaataaagaa agaataaaaa
2881 aagataaaaa gaatagatcc cagccctgtg tataactcac tactttagtc agttccgcag
2941 tattacaaaa ggatgtcgca aacgctgttt gctcctctac aaaacagacc ttaaaaccct
3001 aaaggcttaa gtagcaccct cgcaagctcg gttgcggccg caatcgggca aatcgctgaa
3061 tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt
3121 tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgccttttat
3181 ggattcatgc aaggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc
3241 gttttatggc gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc
3301 cctctgattt tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct
3361 aatgcaccca gtaaggcagc ggtatcatca acggggtctg acgctcagtg gaacgaaaac
3421 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
3481 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
3541 tacgtttcca caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
3601 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
3661 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
3721 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
3781 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
3841 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
3901 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
3961 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
4021 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
4081 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
4141 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
4201 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
4261 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
4321 aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc cgttgaatat
4381 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
4441 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttccc
4501 tgcagggttt gcagtcagag tagaatagaa gtatcaaaaa aagcaccgac tcggtgccac
4561 tttttcaagt tgataacgga ctagccttat tttaacttgc tatgctgttt tgaatggttc
4621 caacaagatt attttataac ttttataaca aataatcaag gagaaattca aagaaattta
4681 tcagccataa aacaatactt aatactatag aatgataaca aaataaacta ctttttaaaa
4741 gaattttgtg ttataatcta tttattatta agtattgggt aatatttttt gaagagatat
4801 tttgaaaaag aaaaattaaa gcatattaaa ctaatttcgg aggtcattaa aactattatt
4861 gaaatcatca aactcattat ggatttaatt taaacttttt attttaggag gcaaaaatgg
4921 ataagaaata ctcaataggc ttagatatcg gcacaaatag cgtcggatgg gcggtgatca
4981 ctgatgatta taaggttccg tctaaaaagt tcaaggttct gggaaataca gaccgccaca
5041 gtatcaaaaa aaatcttata ggggctcttt tatttgacag tggagagaca gcggaagcga
5101 ctcgtctcaa acggacagct cgtagaaggt atacacgtcg gaagaatcgt atttgttatc
5161 tacaggagat tttttcaaat gagatggcga aagtagatga tagtttcttt catcgacttg
5221 aagagtcttt tttggtggaa gaagacaaga agcatgaacg tcatcctatt tttggaaata
5281 tagtagatga agttgcttat catgagaaat atccaactat ctatcatctg cgaaaaaaat
5341 tggtagattc tactgataaa gcggatttgc gcttaatcta tttggcctta gcgcatatga
5401 ttaagtttcg tggtcatttt ttgattgagg gagatttaaa tcctgataat agtgatgtgg
5461 acaaactatt tatccagttg gtacaaacct acaatcaatt atttgaagaa aaccctatta
5521 acgcaagtgg agtagatgct aaagcgattc tttctgcacg attgagtaaa tcaagacgat
5581 tagaaaatct cattgctcag ctccccggtg agaagaaaaa tggcttattt gggaatctca
5641 ttgctttgtc attgggtttg acccctaatt ttaaatcaaa ttttgatttg gcagaagatg
5701 ctaaattaca gctttcaaaa gatacttacg atgatgattt agataattta ttggcgcaaa
5761 ttggagatca atatgctgat ttgtttttgg cagctaagaa tttatcagat gctattttac
5821 tttcagatat cctaagagta aatactgaaa taactaaggc tcccctatca gcttcaatga
5881 ttaaacgcta cgatgaacat catcaagact tgactctttt aaaagcttta gttcgacaac
5941 aacttccaga aaagtataaa gaaatctttt ttgatcaatc aaaaaacgga tatgcaggtt
6001 atattgatgg gggagctagc caagaagaat tttataaatt tatcaaacca attttagaaa
6061 aaatggatgg tactgaggaa ttattggtga aactaaatcg tgaagatttg ctgcgcaagc
6121 aacggacctt tgacaacggc tctattcccc atcaaattca cttgggtgag ctgcatgcta
6181 ttttgagaag acaagaagac ttttatccat ttttaaaaga caatcgtgag aagattgaaa
6241 aaatcttgac ttttcgaatt ccttattatg ttggtccatt ggcgcgtggc aatagtcgtt
6301 ttgcatggat gactcggaag tctgaagaaa caattacccc atggaatttt gaagaagttg
6361 tcgataaagg tgcttcagct caatcattta ttgaacgcat gacaaacttt gataaaaatc
6421 ttccaaatga aaaagtacta ccaaaacata gtttgcttta tgagtatttt acggtttata
6481 acgaattgac aaaggtcaaa tatgttactg aaggaatgcg aaaaccagca tttctttcag
6541 gtgaacagaa gaaagccatt gttgatttac tcttcaaaac aaatcgaaaa gtaaccgtta
6601 agcaattaaa agaagattat ttcaaaaaaa tagaatgttt tgatagtgtt gaaatttcag
6661 gagttgaaga tagatttaat gcttcattag gtacctacca tgatttgcta aaaattatta
6721 aagataaaga ttttttggat aatgaagaaa atgaagatat cttagaggat attgttttaa
6781 cattgacctt atttgaagat agggagatga ttgaggaaag acttaaaaca tatgctcacc
6841 tctttgatga taaggtgatg aaacagctta aacgtcgccg ttatactggt tggggacgtt
6901 tgtctcgaaa attgattaat ggtattaggg ataagcaatc tggcaaaaca atattagatt
6961 ttttgaaatc agatggtttt gccaatcgca attttatgca gctgatccat gatgatagtt
7021 tgacatttaa agaagacatt caaaaagcac aagtgtctgg acaaggcgat agtttacatg
7081 aacatattgc aaatttagct ggtagccctg ctattaaaaa aggtatttta cagactgtaa
7141 aagttgttga tgaattggtc aaagtaatgg ggcggcataa gccagaaaat atcgttattg
7201 aaatggcacg tgaaaatcag acaactcaaa agggccagaa aaattcgcga gagcgtatga
7261 aacgaatcga agaaggtatc aaagaattag gaagtcagat tcttaaagag catcctgttg
7321 aaaatactca attgcaaaat gaaaagctct atctctatta tctccaaaat ggaagagaca
7381 tgtatgtgga ccaagaatta gatattaatc gtttaagtga ttatgatgtc gatcacattg
7441 ttccacaaag tttccttaaa gacgattcaa tagacaataa ggtcttaacg cgttctgata
7501 aaaatcgtgg taaatcggat aacgttccaa gtgaagaagt agtcaaaaag atgaaaaact
7561 attggagaca acttctaaac gccaagttaa tcactcaacg taagtttgat aatttaacga
7621 aagctgaacg tggaggtttg agtgaacttg ataaagctgg ttttatcaaa cgccaattgg
7681 ttgaaactcg ccaaatcact aagcatgtgg cacaaatttt ggatagtcgc atgaatacta
7741 aatacgatga aaatgataaa cttattcgag aggttaaagt gattacctta aaatctaaat
7801 tagtttctga cttccgaaaa gatttccaat tctataaagt acgtgagatt aacaattacc
7861 atcatgccca tgatgcgtat ctaaatgccg tcgttggaac tgctttgatt aagaaatatc
7921 caaaacttga atcggagttt gtctatggtg attataaagt ttatgatgtt cgtaaaatga
7981 ttgctaagtc tgagcaagaa ataggcaaag caaccgcaaa atatttcttt tactctaata
8041 tcatgaactt cttcaaaaca gaaattacac ttgcaaatgg agagattcgc aaacgccctc
8101 taatcgaaac taatggggaa actggagaaa ttgtctggga taaagggcga gattttgcca
8161 cagtgcgcaa agtattgtcc atgccccaag tcaatattgt caagaaaaca gaagtacaga
8221 caggcggatt ctccaaggag tcaattttac caaaaagaaa ttcggacaag cttattgctc
8281 gtaaaaaaga ctgggatcca aaaaaatatg gtggttttga tagtccaacg gtagcttatt
8341 cagtcctagt ggttgctaag gtggaaaaag ggaaatcgaa gaagttaaaa tccgttaaag
8401 agttactagg gatcacaatt atggaaagaa gttcctttga aaaaaatccg attgactttt
8461 tagaagctaa aggatataag gaagttaaaa aagacttaat cattaaacta cctaaatata
8521 gtctttttga gttagaaaac ggtcgtaaac ggatgctggc tagtgccgga gaattacaaa
8581 aaggaaatga gctggctctg ccaagcaaat atgtgaattt tttatattta gctagtcatt
8641 atgaaaagtt gaagggtagt ccagaagata acgaacaaaa acaattgttt gtggagcagc
8701 ataagcatta tttagatgag attattgagc aaatcagtga attttctaag cgtgttattt
8761 tagcagatgc caatttagat aaagttctta gtgcatataa caaacataga gacaaaccaa
8821 tacgtgaaca agcagaaaat attattcatt tatttacgtt gacgaatctt ggagctcccg
8881 ctgcttttaa atattttgat acaacaattg atcgtaaacg atatacgtct acaaaagaag
8941 ttttagatgc cactcttatc catcaatcca tcactggtct ttatgaaaca cgcattgatt
9001 tgagtcagct aggaggtgac tgaagtatat tttagatgaa gattatttct taatctagac
9061 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
9121 tttccccgaa aagtgccacc tgcatcgatt tattatgaca acttgacggc tacatcattc
9181 actttttctt cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat
9241 acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc
9301 atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt
9361 aagacgctaa tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa
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文献和实验1.常用载体分类：2,常用载体元件介绍：(1) 启动子：启动子是位于结构基因5'端上游的DNA序列，能活化RNA聚合酶，使之与模板DNA准确的结合并具有转录起始的特异性。启动子主要分为广谱启动子和特异性启动子。广谱启动子为在各种组织、细胞上广泛表达的启动子；特异性启动子为在特异性组织或者细胞类型上表达的启动子。常用广谱启动子有CAG，CMV，EF1a，PGK等，还包含启动RNA的III型启动子U6和H1启动子；常用特异性启动子如成熟神经元特异性启动子hSyn，投射神经元特异性启动
恰当的选择固相载体和连接分子是固相反应的重要组成部分. 固相载体和连接分子都应该在多种遗传反应过程期间是保持稳定, 而且产品的切除应该是在温和的条件下实现. 本章将介绍各种不同的载体和连接分子. 如果反应后可以用过滤的方法去除, 从理论上讲, 任何材料都可用作固相载体. 聚乙烯, 聚苯乙烯-外涂聚乙烯薄层, 平滑的纸或棉花等都可以. 最常用的载体是: 聚苯乙烯(PS), 聚乙二醇(PEG), Tentagel (TG), 和PEGA (聚乙二醇-聚丙
zhangyuxianggx 各位大侠好！现有pcDNA3-mCherry和pGEM-hgene两个东西在滤纸上，老师就说让构建载体，具体是什么意思啊？我理解的是一个真核表达载体和构建好的pGEM质粒，但是不知下一步该怎么做？恳请各位好心的大侠指点一二。 shylook 直接插入cDNA吧 zhangyuxianggx 那个pGEM就是已经插入了DNA的载体。请问这两个东西溶下来以后
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