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載體基本信息
| Invitrogen |
| pPIC3.5K, pPIC 3.5K |
| 畢赤酵母蛋白表達載體 |
| 高拷貝 |
| AOX1 |
| 多克隆位點，限制性內(nèi)切酶 |
| 9004 bp |
| 5´ AOX1:5´-GACTGGTTCCAATTGACAAGC-3´ |
| 3´ AOX1:5´-GCAAATGGCATTCTGACATCC-3´ |
| / |
| Ampicillin |
| HIS4 |
插入基因是必需包含起始密碼子ATG。 |
| K173-20 |
| 穩(wěn)定 Stable |
| 組成型 Constitutive |
| 非病毒 |
載體質(zhì)粒圖譜和多克隆位點信息
載體簡介
pPIC3.5K是一個鑒定多拷貝基因插入到畢赤酵母基因組中的載體。
1. pPIC3.5K載體大小為9004 bp。
2. 在多克隆位點區(qū)域，擁有5個單一的限制性酶切位點：BamH I，Snab I，EcoR I，Avr II和Not I。
3.胞內(nèi)表達。
4.載體構建時，需要包含一個ATG啟動子，同時需要加入Kozak序列。 (Cavener and Stuart, 1991; Kozak, 1987; Kozak, 1990)
5. 畢赤酵母中使用HIS4基因篩選。
6. 將基因插入GS115和KM71的AOX1中，需要使用Sac I線性化質(zhì)粒(GS115產(chǎn)生的基因型為His+ Mut+，KM71酵母產(chǎn)生的基因型為His+ MutS)。
7. 將基因插入GS115和KM71的HIS4中，需要使用Sal I線性化質(zhì)粒(GS115產(chǎn)生的基因型為His+ Mut+，KM71酵母產(chǎn)生的基因型為His+ MutS)。
8. 將基因插入GS115的AOX1基因中，也可以使用Bgl II 限制酶線性化質(zhì)粒，產(chǎn)生的酵母基因型為His+ MutS。
載體序列
LOCUS pPIC3.5K 9004 bp DNA circular SYN
DEFINITION pPIC3.5K
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
source 1..9004
/organism="pPIC3.5K"
/mol_type="other DNA"
misc_feature complement(1055..1075)
/label="3_AOX1_primer"
CDS complement(1708..4242)
/label="ORF frame 2"
/translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQST
ELQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYR
TLVDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGII
PIIDARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSS
KKSIPEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGF
CHLERTSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAE
AKSKEDIAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQ
PKEESKPKEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQ
NGDKALLELTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLT
ETLQVETCPGVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASP
PKKDGTLTPEVIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTA
AKMMVQNDTSALCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAV
DMTDKELARIEDAVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLI
LQIENASSYVDQVQHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQ
KFITSQDVTPEGLKHIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI*"
gene complement(4656..5471)
/label="KanR2"
/gene="KanR2"
/translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQST
ELQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYR
TLVDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGII
PIIDARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSS
KKSIPEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGF
CHLERTSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAE
AKSKEDIAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQ
PKEESKPKEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQ
NGDKALLELTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLT
ETLQVETCPGVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASP
PKKDGTLTPEVIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTA
AKMMVQNDTSALCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAV
DMTDKELARIEDAVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLI
LQIENASSYVDQVQHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQ
KFITSQDVTPEGLKHIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI*"
CDS complement(4656..5471)
/label="ORF frame 3"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
misc_feature complement(5636..5655)
/label="pBRrevBam_primer"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
CDS complement(5637..6359)
/label="ORF frame 3"
/translation="MTSKHEHTIVSFKTALAWINSRQTTLISLDSEWFERNSNVVTEV
GISIYKPYIPLEGPEGKHLPAPFLPDISTLHFIVQGTEKVRNKKFVPDNKDFFIGGTS
FTVSKKDISAVITEIVSQFESTDDQKSEKFTMKSPPPVALVGHNLIGDLKTLKNAGIT
IPILPIIDTRRSPLCDSCIRKQPSSRLRPLSTAAARNGACKEMAPNSPPATGPATIPT
PKQALMSPKWRARSSPSVMSAI*"
misc_feature complement(6641..6663)
/label="pGEX_3_primer"
/translation="MTSKHEHTIVSFKTALAWINSRQTTLISLDSEWFERNSNVVTEV
GISIYKPYIPLEGPEGKHLPAPFLPDISTLHFIVQGTEKVRNKKFVPDNKDFFIGGTS
FTVSKKDISAVITEIVSQFESTDDQKSEKFTMKSPPPVALVGHNLIGDLKTLKNAGIT
IPILPIIDTRRSPLCDSCIRKQPSSRLRPLSTAAARNGACKEMAPNSPPATGPATIPT
PKQALMSPKWRARSSPSVMSAI*"
rep_origin complement(7060..7679)
/label="pBR322_origin"
/translation="MTSKHEHTIVSFKTALAWINSRQTTLISLDSEWFERNSNVVTEV
GISIYKPYIPLEGPEGKHLPAPFLPDISTLHFIVQGTEKVRNKKFVPDNKDFFIGGTS
FTVSKKDISAVITEIVSQFESTDDQKSEKFTMKSPPPVALVGHNLIGDLKTLKNAGIT
IPILPIIDTRRSPLCDSCIRKQPSSRLRPLSTAAARNGACKEMAPNSPPATGPATIPT
PKQALMSPKWRARSSPSVMSAI*"
gene complement(7834..8694)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MTSKHEHTIVSFKTALAWINSRQTTLISLDSEWFERNSNVVTEV
GISIYKPYIPLEGPEGKHLPAPFLPDISTLHFIVQGTEKVRNKKFVPDNKDFFIGGTS
FTVSKKDISAVITEIVSQFESTDDQKSEKFTMKSPPPVALVGHNLIGDLKTLKNAGIT
IPILPIIDTRRSPLCDSCIRKQPSSRLRPLSTAAARNGACKEMAPNSPPATGPATIPT
PKQALMSPKWRARSSPSVMSAI*"
CDS complement(7834..8694)
/label="ORF frame 2"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(8736..8764)
/label="AmpR_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
ORIGIN
1 AGATCTAACA TCCAAAGACG AAAGGTTGAA TGAAACCTTT TTGCCATCCG ACATCCACAG
61 GTCCATTCTC ACACATAAGT GCCAAACGCA ACAGGAGGGG ATACACTAGC AGCAGACCGT
121 TGCAAACGCA GGACCTCCAC TCCTCTTCTC CTCAACACCC ACTTTTGCCA TCGAAAAACC
181 AGCCCAGTTA TTGGGCTTGA TTGGAGCTCG CTCATTCCAA TTCCTTCTAT TAGGCTACTA
241 ACACCATGAC TTTATTAGCC TGTCTATCCT GGCCCCCCTG GCGAGGTTCA TGTTTGTTTA
301 TTTCCGAATG CAACAAGCTC CGCATTACAC CCGAACATCA CTCCAGATGA GGGCTTTCTG
361 AGTGTGGGGT CAAATAGTTT CATGTTCCCC AAATGGCCCA AAACTGACAG TTTAAACGCT
421 GTCTTGGAAC CTAATATGAC AAAAGCGTGA TCTCATCCAA GATGAACTAA GTTTGGTTCG
481 TTGAAATGCT AACGGCCAGT TGGTCAAAAA GAAACTTCCA AAAGTCGCCA TACCGTTTGT
541 CTTGTTTGGT ATTGATTGAC GAATGCTCAA AAATAATCTC ATTAATGCTT AGCGCAGTCT
601 CTCTATCGCT TCTGAACCCC GGTGCACCTG TGCCGAAACG CAAATGGGGA AACACCCGCT
661 TTTTGGATGA TTATGCATTG TCTCCACATT GTATGCTTCC AAGATTCTGG TGGGAATACT
721 GCTGATAGCC TAACGTTCAT GATCAAAATT TAACTGTTCT AACCCCTACT TGACAGCAAT
781 ATATAAACAG AAGGAAGCTG CCCTGTCTTA AACCTTTTTT TTTATCATCA TTATTAGCTT
841 ACTTTCATAA TTGCGACTGG TTCCAATTGA CAAGCTTTTG ATTTTAACGA CTTTTAACGA
901 CAACTTGAGA AGATCAAAAA ACAACTAATT ATTCGAAGGA TCCTACGTAG AATTCCCTAG
961 GGCGGCCGCG AATTAATTCG CCTTAGACAT GACTGTTCCT CAGTTCAAGT TGGGCACTTA
1021 CGAGAAGACC GGTCTTGCTA GATTCTAATC AAGAGGATGT CAGAATGCCA TTTGCCTGAG
1081 AGATGCAGGC TTCATTTTTG ATACTTTTTT ATTTGTAACC TATATAGTAT AGGATTTTTT
1141 TTGTCATTTT GTTTCTTCTC GTACGAGCTT GCTCCTGATC AGCCTATCTC GCAGCTGATG
1201 AATATCTTGT GGTAGGGGTT TGGGAAAATC ATTCGAGTTT GATGTTTTTC TTGGTATTTC
1261 CCACTCCTCT TCAGAGTACA GAAGATTAAG TGAGAAGTTC GTTTGTGCAA GCTTATCGAT
1321 AAGCTTTAAT GCGGTAGTTT ATCACAGTTA AATTGCTAAC GCAGTCAGGC ACCGTGTATG
1381 AAATCTAACA ATGCGCTCAT CGTCATCCTC GGCACCGTCA CCCTGGATGC TGTAGGCATA
1441 GGCTTGGTTA TGCCGGTACT GCCGGGCCTC TTGCGGGATA TCGTCCATTC CGACAGCATC
1501 GCCAGTCACT ATGGCGTGCT GCTAGCGCTA TATGCGTTGA TGCAATTTCT ATGCGCACCC
1561 GTTCTCGGAG CACTGTCCGA CCGCTTTGGC CGCCGCCCAG TCCTGCTCGC TTCGCTACTT
1621 GGAGCCACTA TCGACTACGC GATCATGGCG ACCACACCCG TCCTGTGGAT CTATCGAATC
1681 TAAATGTAAG TTAAAATCTC TAAATAATTA AATAAGTCCC AGTTTCTCCA TACGAACCTT
1741 AACAGCATTG CGGTGAGCAT CTAGACCTTC AACAGCAGCC AGATCCATCA CTGCTTGGCC
1801 AATATGTTTC AGTCCCTCAG GAGTTACGTC TTGTGAAGTG ATGAACTTCT GGAAGGTTGC
1861 AGTGTTAACT CCGCTGTATT GACGGGCATA TCCGTACGTT GGCAAAGTGT GGTTGGTACC
1921 GGAGGAGTAA TCTCCACAAC TCTCTGGAGA GTAGGCACCA ACAAACACAG ATCCAGCGTG
1981 TTGTACTTGA TCAACATAAG AAGAAGCATT CTCGATTTGC AGGATCAAGT GTTCAGGAGC
2041 GTACTGATTG GACATTTCCA AAGCCTGCTC GTAGGTTGCA ACCGATAGGG TTGTAGAGTG
2101 TGCAATACAC TTGCGTACAA TTTCAACCCT TGGCAACTGC ACAGCTTGGT TGTGAACAGC
2161 ATCTTCAATT CTGGCAAGCT CCTTGTCTGT CATATCGACA GCCAACAGAA TCACCTGGGA
2221 ATCAATACCA TGTTCAGCTT GAGACAGAAG GTCTGAGGCA ACGAAATCTG GATCAGCGTA
2281 TTTATCAGCA ATAACTAGAA CTTCAGAAGG CCCAGCAGGC ATGTCAATAC TACACAGGGC
2341 TGATGTGTCA TTTTGAACCA TCATCTTGGC AGCAGTAACG AACTGGTTTC CTGGACCAAA
2401 TATTTTGTCA CACTTAGGAA CAGTTTCTGT TCCGTAAGCC ATAGCAGCTA CTGCCTGGGC
2461 GCCTCCTGCT AGCACGATAC ACTTAGCACC AACCTTGTGG GCAACGTAGA TGACTTCTGG
2521 GGTAAGGGTA CCATCCTTCT TAGGTGGAGA TGCAAAAACA ATTTCTTTGC AACCAGCAAC
2581 TTTGGCAGGA ACACCCAGCA TCAGGGAAGT GGAAGGCAGA ATTGCGGTTC CACCAGGAAT
2641 ATAGAGGCCA ACTTTCTCAA TAGGTCTTGC AAAACGAGAG CAGACTACAC CAGGGCAAGT
2701 CTCAACTTGC AACGTCTCCG TTAGTTGAGC TTCATGGAAT TTCCTGACGT TATCTATAGA
2761 GAGATCAATG GCTCTCTTAA CGTTATCTGG CAATTGCATA AGTTCCTCTG GGAAAGGAGC
2821 TTCTAACACA GGTGTCTTCA AAGCGACTCC ATCAAACTTG GCAGTTAGTT CTAAAAGGGC
2881 TTTGTCACCA TTTTGACGAA CATTGTCGAC AATTGGTTTG ACTAATTCCA TAATCTGTTC
2941 CGTTTTCTGG ATAGGACGAC GAAGGGCATC TTCAATTTCT TGTGAGGAGG CCTTAGAAAC
3001 GTCAATTTTG CACAATTCAA TACGACCTTC AGAAGGGACT TCTTTAGGTT TGGATTCTTC
3061 TTTAGGTTGT TCCTTGGTGT ATCCTGGCTT GGCATCTCCT TTCCTTCTAG TGACCTTTAG
3121 GGACTTCATA TCCAGGTTTC TCTCCACCTC GTCCAACGTC ACACCGTACT TGGCACATCT
3181 AACTAATGCA AAATAAAATA AGTCAGCACA TTCCCAGGCT ATATCTTCCT TGGATTTAGC
3241 TTCTGCAAGT TCATCAGCTT CCTCCCTAAT TTTAGCGTTC AACAAAACTT CGTCGTCAAA
3301 TAACCGTTTG GTATAAGAAC CTTCTGGAGC ATTGCTCTTA CGATCCCACA AGGTGGCTTC
3361 CATGGCTCTA AGACCCTTTG ATTGGCCAAA ACAGGAAGTG CGTTCCAAGT GACAGAAACC
3421 AACACCTGTT TGTTCAACCA CAAATTTCAA GCAGTCTCCA TCACAATCCA ATTCGATACC
3481 CAGCAACTTT TGAGTTGCTC CAGATGTAGC ACCTTTATAC CACAAACCGT GACGACGAGA
3541 TTGGTAGACT CCAGTTTGTG TCCTTATAGC CTCCGGAATA GACTTTTTGG ACGAGTACAC
3601 CAGGCCCAAC GAGTAATTAG AAGAGTCAGC CACCAAAGTA GTGAATAGAC CATCGGGGCG
3661 GTCAGTAGTC AAAGACGCCA ACAAAATTTC ACTGACAGGG AACTTTTTGA CATCTTCAGA
3721 AAGTTCGTAT TCAGTAGTCA ATTGCCGAGC ATCAATAATG GGGATTATAC CAGAAGCAAC
3781 AGTGGAAGTC ACATCTACCA ACTTTGCGGT CTCAGAAAAA GCATAAACAG TTCTACTACC
3841 GCCATTAGTG AAACTTTTCA AATCGCCCAG TGGAGAAGAA AAAGGCACAG CGATACTAGC
3901 ATTAGCGGGC AAGGATGCAA CTTTATCAAC CAGGGTCCTA TAGATAACCC TAGCGCCTGG
3961 GATCATCCTT TGGACAACTC TTTCTGCCAA ATCTAGGTCC AAAATCACTT CATTGATACC
4021 ATTATTGTAC AACTTGAGCA AGTTGTCGAT CAGCTCCTCA AATTGGTCCT CTGTAACGGA
4081 TGACTCAACT TGCACATTAA CTTGAAGCTC AGTCGATTGA GTGAACTTGA TCAGGTTGTG
4141 CAGCTGGTCA GCAGCATAGG GAAACACGGC TTTTCCTACC AAACTCAAGG AATTATCAAA
4201 CTCTGCAACA CTTGCGTATG CAGGTAGCAA GGGAAATGTC ATACTTGAAG TCGGACAGTG
4261 AGTGTAGTCT TGAGAAATTC TGAAGCCGTA TTTTTATTAT CAGTGAGTCA GTCATCAGGA
4321 GATCCTCTAC GCCGGACGCA TCGTGGCCGA CCTGCAGGGG GGGGGGGGGC GCTGAGGTCT
4381 GCCTCGTGAA GAAGGTGTTG CTGACTCATA CCAGGCCTGA ATCGCCCCAT CATCCAGCCA
4441 GAAAGTGAGG GAGCCACGGT TGATGAGAGC TTTGTTGTAG GTGGACCAGT TGGTGATTTT
4501 GAACTTTTGC TTTGCCACGG AACGGTCTGC GTTGTCGGGA AGATGCGTGA TCTGATCCTT
4561 CAACTCAGCA AAAGTTCGAT TTATTCAACA AAGCCGCCGT CCCGTCAAGT CAGCGTAATG
4621 CTCTGCCAGT GTTACAACCA ATTAACCAAT TCTGATTAGA AAAACTCATC GAGCATCAAA
4681 TGAAACTGCA ATTTATTCAT ATCAGGATTA TCAATACCAT ATTTTTGAAA AAGCCGTTTC
4741 TGTAATGAAG GAGAAAACTC ACCGAGGCAG TTCCATAGGA TGGCAAGATC CTGGTATCGG
4801 TCTGCGATTC CGACTCGTCC AACATCAATA CAACCTATTA ATTTCCCCTC GTCAAAAATA
4861 AGGTTATCAA GTGAGAAATC ACCATGAGTG ACGACTGAAT CCGGTGAGAA TGGCAAAAGC
4921 TTATGCATTT CTTTCCAGAC TTGTTCAACA GGCCAGCCAT TACGCTCGTC ATCAAAATCA
4981 CTCGCATCAA CCAAACCGTT ATTCATTCGT GATTGCGCCT GAGCGAGACG AAATACGCGA
5041 TCGCTGTTAA AAGGACAATT ACAAACAGGA ATCGAATGCA ACCGGCGCAG GAACACTGCC
5101 AGCGCATCAA CAATATTTTC ACCTGAATCA GGATATTCTT CTAATACCTG GAATGCTGTT
5161 TTCCCGGGGA TCGCAGTGGT GAGTAACCAT GCATCATCAG GAGTACGGAT AAAATGCTTG
5221 ATGGTCGGAA GAGGCATAAA TTCCGTCAGC CAGTTTAGTC TGACCATCTC ATCTGTAACA
5281 TCATTGGCAA CGCTACCTTT GCCATGTTTC AGAAACAACT CTGGCGCATC GGGCTTCCCA
5341 TACAATCGAT AGATTGTCGC ACCTGATTGC CCGACATTAT CGCGAGCCCA TTTATACCCA
5401 TATAAATCAG CATCCATGTT GGAATTTAAT CG
DEFINITION pPIC3.5K
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
source 1..9004
/organism="pPIC3.5K"
/mol_type="other DNA"
misc_feature complement(1055..1075)
/label="3_AOX1_primer"
CDS complement(1708..4242)
/label="ORF frame 2"
/translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQST
ELQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYR
TLVDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGII
PIIDARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSS
KKSIPEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGF
CHLERTSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAE
AKSKEDIAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQ
PKEESKPKEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQ
NGDKALLELTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLT
ETLQVETCPGVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASP
PKKDGTLTPEVIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTA
AKMMVQNDTSALCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAV
DMTDKELARIEDAVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLI
LQIENASSYVDQVQHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQ
KFITSQDVTPEGLKHIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI*"
gene complement(4656..5471)
/label="KanR2"
/gene="KanR2"
/translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQST
ELQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYR
TLVDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGII
PIIDARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSS
KKSIPEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGF
CHLERTSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAE
AKSKEDIAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQ
PKEESKPKEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQ
NGDKALLELTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLT
ETLQVETCPGVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASP
PKKDGTLTPEVIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTA
AKMMVQNDTSALCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAV
DMTDKELARIEDAVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLI
LQIENASSYVDQVQHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQ
KFITSQDVTPEGLKHIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI*"
CDS complement(4656..5471)
/label="ORF frame 3"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
misc_feature complement(5636..5655)
/label="pBRrevBam_primer"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
CDS complement(5637..6359)
/label="ORF frame 3"
/translation="MTSKHEHTIVSFKTALAWINSRQTTLISLDSEWFERNSNVVTEV
GISIYKPYIPLEGPEGKHLPAPFLPDISTLHFIVQGTEKVRNKKFVPDNKDFFIGGTS
FTVSKKDISAVITEIVSQFESTDDQKSEKFTMKSPPPVALVGHNLIGDLKTLKNAGIT
IPILPIIDTRRSPLCDSCIRKQPSSRLRPLSTAAARNGACKEMAPNSPPATGPATIPT
PKQALMSPKWRARSSPSVMSAI*"
misc_feature complement(6641..6663)
/label="pGEX_3_primer"
/translation="MTSKHEHTIVSFKTALAWINSRQTTLISLDSEWFERNSNVVTEV
GISIYKPYIPLEGPEGKHLPAPFLPDISTLHFIVQGTEKVRNKKFVPDNKDFFIGGTS
FTVSKKDISAVITEIVSQFESTDDQKSEKFTMKSPPPVALVGHNLIGDLKTLKNAGIT
IPILPIIDTRRSPLCDSCIRKQPSSRLRPLSTAAARNGACKEMAPNSPPATGPATIPT
PKQALMSPKWRARSSPSVMSAI*"
rep_origin complement(7060..7679)
/label="pBR322_origin"
/translation="MTSKHEHTIVSFKTALAWINSRQTTLISLDSEWFERNSNVVTEV
GISIYKPYIPLEGPEGKHLPAPFLPDISTLHFIVQGTEKVRNKKFVPDNKDFFIGGTS
FTVSKKDISAVITEIVSQFESTDDQKSEKFTMKSPPPVALVGHNLIGDLKTLKNAGIT
IPILPIIDTRRSPLCDSCIRKQPSSRLRPLSTAAARNGACKEMAPNSPPATGPATIPT
PKQALMSPKWRARSSPSVMSAI*"
gene complement(7834..8694)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MTSKHEHTIVSFKTALAWINSRQTTLISLDSEWFERNSNVVTEV
GISIYKPYIPLEGPEGKHLPAPFLPDISTLHFIVQGTEKVRNKKFVPDNKDFFIGGTS
FTVSKKDISAVITEIVSQFESTDDQKSEKFTMKSPPPVALVGHNLIGDLKTLKNAGIT
IPILPIIDTRRSPLCDSCIRKQPSSRLRPLSTAAARNGACKEMAPNSPPATGPATIPT
PKQALMSPKWRARSSPSVMSAI*"
CDS complement(7834..8694)
/label="ORF frame 2"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(8736..8764)
/label="AmpR_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
ORIGIN
1 AGATCTAACA TCCAAAGACG AAAGGTTGAA TGAAACCTTT TTGCCATCCG ACATCCACAG
61 GTCCATTCTC ACACATAAGT GCCAAACGCA ACAGGAGGGG ATACACTAGC AGCAGACCGT
121 TGCAAACGCA GGACCTCCAC TCCTCTTCTC CTCAACACCC ACTTTTGCCA TCGAAAAACC
181 AGCCCAGTTA TTGGGCTTGA TTGGAGCTCG CTCATTCCAA TTCCTTCTAT TAGGCTACTA
241 ACACCATGAC TTTATTAGCC TGTCTATCCT GGCCCCCCTG GCGAGGTTCA TGTTTGTTTA
301 TTTCCGAATG CAACAAGCTC CGCATTACAC CCGAACATCA CTCCAGATGA GGGCTTTCTG
361 AGTGTGGGGT CAAATAGTTT CATGTTCCCC AAATGGCCCA AAACTGACAG TTTAAACGCT
421 GTCTTGGAAC CTAATATGAC AAAAGCGTGA TCTCATCCAA GATGAACTAA GTTTGGTTCG
481 TTGAAATGCT AACGGCCAGT TGGTCAAAAA GAAACTTCCA AAAGTCGCCA TACCGTTTGT
541 CTTGTTTGGT ATTGATTGAC GAATGCTCAA AAATAATCTC ATTAATGCTT AGCGCAGTCT
601 CTCTATCGCT TCTGAACCCC GGTGCACCTG TGCCGAAACG CAAATGGGGA AACACCCGCT
661 TTTTGGATGA TTATGCATTG TCTCCACATT GTATGCTTCC AAGATTCTGG TGGGAATACT
721 GCTGATAGCC TAACGTTCAT GATCAAAATT TAACTGTTCT AACCCCTACT TGACAGCAAT
781 ATATAAACAG AAGGAAGCTG CCCTGTCTTA AACCTTTTTT TTTATCATCA TTATTAGCTT
841 ACTTTCATAA TTGCGACTGG TTCCAATTGA CAAGCTTTTG ATTTTAACGA CTTTTAACGA
901 CAACTTGAGA AGATCAAAAA ACAACTAATT ATTCGAAGGA TCCTACGTAG AATTCCCTAG
961 GGCGGCCGCG AATTAATTCG CCTTAGACAT GACTGTTCCT CAGTTCAAGT TGGGCACTTA
1021 CGAGAAGACC GGTCTTGCTA GATTCTAATC AAGAGGATGT CAGAATGCCA TTTGCCTGAG
1081 AGATGCAGGC TTCATTTTTG ATACTTTTTT ATTTGTAACC TATATAGTAT AGGATTTTTT
1141 TTGTCATTTT GTTTCTTCTC GTACGAGCTT GCTCCTGATC AGCCTATCTC GCAGCTGATG
1201 AATATCTTGT GGTAGGGGTT TGGGAAAATC ATTCGAGTTT GATGTTTTTC TTGGTATTTC
1261 CCACTCCTCT TCAGAGTACA GAAGATTAAG TGAGAAGTTC GTTTGTGCAA GCTTATCGAT
1321 AAGCTTTAAT GCGGTAGTTT ATCACAGTTA AATTGCTAAC GCAGTCAGGC ACCGTGTATG
1381 AAATCTAACA ATGCGCTCAT CGTCATCCTC GGCACCGTCA CCCTGGATGC TGTAGGCATA
1441 GGCTTGGTTA TGCCGGTACT GCCGGGCCTC TTGCGGGATA TCGTCCATTC CGACAGCATC
1501 GCCAGTCACT ATGGCGTGCT GCTAGCGCTA TATGCGTTGA TGCAATTTCT ATGCGCACCC
1561 GTTCTCGGAG CACTGTCCGA CCGCTTTGGC CGCCGCCCAG TCCTGCTCGC TTCGCTACTT
1621 GGAGCCACTA TCGACTACGC GATCATGGCG ACCACACCCG TCCTGTGGAT CTATCGAATC
1681 TAAATGTAAG TTAAAATCTC TAAATAATTA AATAAGTCCC AGTTTCTCCA TACGAACCTT
1741 AACAGCATTG CGGTGAGCAT CTAGACCTTC AACAGCAGCC AGATCCATCA CTGCTTGGCC
1801 AATATGTTTC AGTCCCTCAG GAGTTACGTC TTGTGAAGTG ATGAACTTCT GGAAGGTTGC
1861 AGTGTTAACT CCGCTGTATT GACGGGCATA TCCGTACGTT GGCAAAGTGT GGTTGGTACC
1921 GGAGGAGTAA TCTCCACAAC TCTCTGGAGA GTAGGCACCA ACAAACACAG ATCCAGCGTG
1981 TTGTACTTGA TCAACATAAG AAGAAGCATT CTCGATTTGC AGGATCAAGT GTTCAGGAGC
2041 GTACTGATTG GACATTTCCA AAGCCTGCTC GTAGGTTGCA ACCGATAGGG TTGTAGAGTG
2101 TGCAATACAC TTGCGTACAA TTTCAACCCT TGGCAACTGC ACAGCTTGGT TGTGAACAGC
2161 ATCTTCAATT CTGGCAAGCT CCTTGTCTGT CATATCGACA GCCAACAGAA TCACCTGGGA
2221 ATCAATACCA TGTTCAGCTT GAGACAGAAG GTCTGAGGCA ACGAAATCTG GATCAGCGTA
2281 TTTATCAGCA ATAACTAGAA CTTCAGAAGG CCCAGCAGGC ATGTCAATAC TACACAGGGC
2341 TGATGTGTCA TTTTGAACCA TCATCTTGGC AGCAGTAACG AACTGGTTTC CTGGACCAAA
2401 TATTTTGTCA CACTTAGGAA CAGTTTCTGT TCCGTAAGCC ATAGCAGCTA CTGCCTGGGC
2461 GCCTCCTGCT AGCACGATAC ACTTAGCACC AACCTTGTGG GCAACGTAGA TGACTTCTGG
2521 GGTAAGGGTA CCATCCTTCT TAGGTGGAGA TGCAAAAACA ATTTCTTTGC AACCAGCAAC
2581 TTTGGCAGGA ACACCCAGCA TCAGGGAAGT GGAAGGCAGA ATTGCGGTTC CACCAGGAAT
2641 ATAGAGGCCA ACTTTCTCAA TAGGTCTTGC AAAACGAGAG CAGACTACAC CAGGGCAAGT
2701 CTCAACTTGC AACGTCTCCG TTAGTTGAGC TTCATGGAAT TTCCTGACGT TATCTATAGA
2761 GAGATCAATG GCTCTCTTAA CGTTATCTGG CAATTGCATA AGTTCCTCTG GGAAAGGAGC
2821 TTCTAACACA GGTGTCTTCA AAGCGACTCC ATCAAACTTG GCAGTTAGTT CTAAAAGGGC
2881 TTTGTCACCA TTTTGACGAA CATTGTCGAC AATTGGTTTG ACTAATTCCA TAATCTGTTC
2941 CGTTTTCTGG ATAGGACGAC GAAGGGCATC TTCAATTTCT TGTGAGGAGG CCTTAGAAAC
3001 GTCAATTTTG CACAATTCAA TACGACCTTC AGAAGGGACT TCTTTAGGTT TGGATTCTTC
3061 TTTAGGTTGT TCCTTGGTGT ATCCTGGCTT GGCATCTCCT TTCCTTCTAG TGACCTTTAG
3121 GGACTTCATA TCCAGGTTTC TCTCCACCTC GTCCAACGTC ACACCGTACT TGGCACATCT
3181 AACTAATGCA AAATAAAATA AGTCAGCACA TTCCCAGGCT ATATCTTCCT TGGATTTAGC
3241 TTCTGCAAGT TCATCAGCTT CCTCCCTAAT TTTAGCGTTC AACAAAACTT CGTCGTCAAA
3301 TAACCGTTTG GTATAAGAAC CTTCTGGAGC ATTGCTCTTA CGATCCCACA AGGTGGCTTC
3361 CATGGCTCTA AGACCCTTTG ATTGGCCAAA ACAGGAAGTG CGTTCCAAGT GACAGAAACC
3421 AACACCTGTT TGTTCAACCA CAAATTTCAA GCAGTCTCCA TCACAATCCA ATTCGATACC
3481 CAGCAACTTT TGAGTTGCTC CAGATGTAGC ACCTTTATAC CACAAACCGT GACGACGAGA
3541 TTGGTAGACT CCAGTTTGTG TCCTTATAGC CTCCGGAATA GACTTTTTGG ACGAGTACAC
3601 CAGGCCCAAC GAGTAATTAG AAGAGTCAGC CACCAAAGTA GTGAATAGAC CATCGGGGCG
3661 GTCAGTAGTC AAAGACGCCA ACAAAATTTC ACTGACAGGG AACTTTTTGA CATCTTCAGA
3721 AAGTTCGTAT TCAGTAGTCA ATTGCCGAGC ATCAATAATG GGGATTATAC CAGAAGCAAC
3781 AGTGGAAGTC ACATCTACCA ACTTTGCGGT CTCAGAAAAA GCATAAACAG TTCTACTACC
3841 GCCATTAGTG AAACTTTTCA AATCGCCCAG TGGAGAAGAA AAAGGCACAG CGATACTAGC
3901 ATTAGCGGGC AAGGATGCAA CTTTATCAAC CAGGGTCCTA TAGATAACCC TAGCGCCTGG
3961 GATCATCCTT TGGACAACTC TTTCTGCCAA ATCTAGGTCC AAAATCACTT CATTGATACC
4021 ATTATTGTAC AACTTGAGCA AGTTGTCGAT CAGCTCCTCA AATTGGTCCT CTGTAACGGA
4081 TGACTCAACT TGCACATTAA CTTGAAGCTC AGTCGATTGA GTGAACTTGA TCAGGTTGTG
4141 CAGCTGGTCA GCAGCATAGG GAAACACGGC TTTTCCTACC AAACTCAAGG AATTATCAAA
4201 CTCTGCAACA CTTGCGTATG CAGGTAGCAA GGGAAATGTC ATACTTGAAG TCGGACAGTG
4261 AGTGTAGTCT TGAGAAATTC TGAAGCCGTA TTTTTATTAT CAGTGAGTCA GTCATCAGGA
4321 GATCCTCTAC GCCGGACGCA TCGTGGCCGA CCTGCAGGGG GGGGGGGGGC GCTGAGGTCT
4381 GCCTCGTGAA GAAGGTGTTG CTGACTCATA CCAGGCCTGA ATCGCCCCAT CATCCAGCCA
4441 GAAAGTGAGG GAGCCACGGT TGATGAGAGC TTTGTTGTAG GTGGACCAGT TGGTGATTTT
4501 GAACTTTTGC TTTGCCACGG AACGGTCTGC GTTGTCGGGA AGATGCGTGA TCTGATCCTT
4561 CAACTCAGCA AAAGTTCGAT TTATTCAACA AAGCCGCCGT CCCGTCAAGT CAGCGTAATG
4621 CTCTGCCAGT GTTACAACCA ATTAACCAAT TCTGATTAGA AAAACTCATC GAGCATCAAA
4681 TGAAACTGCA ATTTATTCAT ATCAGGATTA TCAATACCAT ATTTTTGAAA AAGCCGTTTC
4741 TGTAATGAAG GAGAAAACTC ACCGAGGCAG TTCCATAGGA TGGCAAGATC CTGGTATCGG
4801 TCTGCGATTC CGACTCGTCC AACATCAATA CAACCTATTA ATTTCCCCTC GTCAAAAATA
4861 AGGTTATCAA GTGAGAAATC ACCATGAGTG ACGACTGAAT CCGGTGAGAA TGGCAAAAGC
4921 TTATGCATTT CTTTCCAGAC TTGTTCAACA GGCCAGCCAT TACGCTCGTC ATCAAAATCA
4981 CTCGCATCAA CCAAACCGTT ATTCATTCGT GATTGCGCCT GAGCGAGACG AAATACGCGA
5041 TCGCTGTTAA AAGGACAATT ACAAACAGGA ATCGAATGCA ACCGGCGCAG GAACACTGCC
5101 AGCGCATCAA CAATATTTTC ACCTGAATCA GGATATTCTT CTAATACCTG GAATGCTGTT
5161 TTCCCGGGGA TCGCAGTGGT GAGTAACCAT GCATCATCAG GAGTACGGAT AAAATGCTTG
5221 ATGGTCGGAA GAGGCATAAA TTCCGTCAGC CAGTTTAGTC TGACCATCTC ATCTGTAACA
5281 TCATTGGCAA CGCTACCTTT GCCATGTTTC AGAAACAACT CTGGCGCATC GGGCTTCCCA
5341 TACAATCGAT AGATTGTCGC ACCTGATTGC CCGACATTAT CGCGAGCCCA TTTATACCCA
5401 TATAAATCAG CATCCATGTT GGAATTTAAT CG
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