| 出品公司: | Invitrogen |
|---|---|
| 载体名称: | pPIC3.5, pPIC 3.5 |
| 质粒类型: | 毕赤酵母蛋白表达载体 |
| 表达水平: | 高拷贝 |
| 启动子: | AOX1 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 7751 bp |
| 5' 测序引物及序列: | 5´ AOX1:5´-GACTGGTTCCAATTGACAAGC-3´ |
| 3' 测序引物及序列: | 3´ AOX1:5´-GCAAATGGCATTCTGACATCC-3´ |
| 载体标签: | / |
| 载体抗性: | Ampicillin |
| 筛选标记: | HIS4 |
| 备注: |
插入基因是必需包含起始密码子ATG。
|
| 产品目录号: | K1710-01 |
| 稳定性: | 稳定 Stable |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
买家导航
pPIC3.5载体质粒基本信息
pPIC3.5质粒图谱载体图谱和pPIC3.5载体序列质粒序列多克隆位点信息
pPIC3.5质粒载体简介
pPIC3.5酵母表达载体详细信息如下:
1. 载体大小为7,751 bp，是非融合蛋白表达载体。
2. 单一限制性内切酶位点有BamH I, SnaB I, EcoR I, Avr II 和Not I.
3. 胞内基因表达载体，不是分泌性蛋白表达载体。
4. 载体构建时，需要包含一个ATG启动子，同时需要加入Kozak序列。 (Cavener and Stuart, 1991; Kozak, 1987; Kozak, 1990)
5. 毕赤酵母中使用HIS4基因筛选。
6. 将基因插入GS115和KM71的AOX1中，需要使用Sac I线性化质粒(GS115产生的基因型为His+ Mut+，KM71酵母产生的基因型为His+ MutS)。
7. 将基因插入GS115和KM71的HIS4中，需要使用Sal I或 Stu I线性化质粒(GS115产生的基因型为His+ Mut+，KM71酵母产生的基因型为His+ MutS)。
8. 将基因插入GS115的AOX1基因中，也可以使用Bgl II 限制酶线性化质粒，产生的酵母基因型为His+ MutS。
pPIC3.5质粒序列载体序列
LOCUS pPIC3.5 7751 bp ds-DNA circular SYN 05-二月-2013
DEFINITION Pichia pastoris HIS4 vector for methanol-inducible intracellular
expression of a protein.
ACCESSION .
VERSION .
KEYWORDS pPIC3.5
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
source 1..7751
/organism="synthetic DNA construct"
/lab_host="Pichia pastoris"
/mol_type="other DNA"
promoter 2..937
/gene="Pichia pastoris AOX1"
/note="AOX1 promoter"
/note="inducible promoter, regulated by methanol"
/note="color: #ffffff; direction: RIGHT"
misc_feature 938..969
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
terminator 1049..1295
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
/note="color: #ffffff"
CDS complement(1708..4242)
/codon_start=1
/gene="Pichia pastoris HIS4"
/locus_tag="
"
/product="multifunctional enzyme, required for histidine
biosynthesis"
/note="PpHIS4"
/note="auxotrophic marker for Pichia pastoris"
/note="color: #ff7f50"
/protein_id="
"
/translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE
VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
misc_feature 4597..5353
/note="AOX1 3' fragment"
/note="region downstream of Pichia pastoris AOX1 gene"
/note="color: #ffffff"
rep_origin complement(5822..6410)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
CDS complement(6581..7441)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/note="This feature has 2 segments:
???1:?6581?..?7372?/?#ccffcc
???2:?7373?..?7441?/?#ccffcc?/?signal?sequence
Cleavage site after base 7372"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7442..7546)
/gene="bla"
/note="AmpR promoter"
/note="color: #ffffff; direction: LEFT"
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 tcgtggccgg catcaccggc gccacaggtg cggttgctgg
4381 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
4441 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
4501 ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg
4561 ctgcttccta atgcaggagt cgcataaggg agagcgtcga gtatctatga ttggaagtat
4621 gggaatggtg atacccgcat tcttcagtgt cttgaggtct cctatcagat tatgcccaac
4681 taaagcaacc ggaggaggag atttcatggt aaatttctct gacttttggt catcagtaga
4741 ctcgaactgt gagactatct cggttatgac agcagaaatg tccttcttgg agacagtaaa
4801 tgaagtccca ccaataaaga aatccttgtt atcaggaaca aacttcttgt ttcgaacttt
4861 ttcggtgcct tgaactataa aatgtagagt ggatatgtcg ggtaggaatg gagcgggcaa
4921 atgcttacct tctggacctt caagaggtat gtagggtttg tagatactga tgccaacttc
4981 agtgacaacg ttgctatttc gttcaaacca ttccgaatcc agagaaatca aagttgtttg
5041 tctactattg atccaagcca gtgcggtctt gaaactgaca atagtgtgct cgtgttttga
5101 ggtcatcttt gtatgaataa atctagtctt tgatctaaat aatcttgacg agccaaggcg
5161 ataaataccc aaatctaaaa ctcttttaaa acgttaaaag gacaagtatg tctgcctgta
5221 ttaaacccca aatcagctcg tagtctgatc ctcatcaact tgaggggcac tatcttgttt
5281 tagagaaatt tgcggagatg cgatatcgag aaaaaggtac gctgatttta aacgtgaaat
5341 ttatctcaag atctctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
5401 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
5461 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
5521 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
5581 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
5641 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
5701 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
5761 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
5821 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
5881 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
5941 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
6001 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
6061 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
6121 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
6181 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
6241 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
6301 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
6361 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
6421 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
6481 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
6541 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
6601 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
6661 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
6721 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
6781 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
6841 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
6901 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
6961 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
7021 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
7081 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
7141 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
7201 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
7261 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
7321 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
7381 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
7441 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
7501 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
7561 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
7621 gtatcacgag gccctttcgt cttcaagaat taattctcat gtttgacagc ttatcatcga
7681 taagctgact catgttggta ttgtgaaata gacgcagatc gggaacactg aaaaataaca
7741 gttattattc g
//
DEFINITION Pichia pastoris HIS4 vector for methanol-inducible intracellular
expression of a protein.
ACCESSION .
VERSION .
KEYWORDS pPIC3.5
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
source 1..7751
/organism="synthetic DNA construct"
/lab_host="Pichia pastoris"
/mol_type="other DNA"
promoter 2..937
/gene="Pichia pastoris AOX1"
/note="AOX1 promoter"
/note="inducible promoter, regulated by methanol"
/note="color: #ffffff; direction: RIGHT"
misc_feature 938..969
/note="MCS"
/note="multiple cloning site"
/note="color: #99ccff"
terminator 1049..1295
/gene="Pichia pastoris AOX1"
/note="AOX1 terminator"
/note="transcription terminator for AOX1"
/note="color: #ffffff"
CDS complement(1708..4242)
/codon_start=1
/gene="Pichia pastoris HIS4"
/locus_tag="
"
/product="multifunctional enzyme, required for histidine
biosynthesis"
/note="PpHIS4"
/note="auxotrophic marker for Pichia pastoris"
/note="color: #ff7f50"
/protein_id="
"
/translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE
VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
misc_feature 4597..5353
/note="AOX1 3' fragment"
/note="region downstream of Pichia pastoris AOX1 gene"
/note="color: #ffffff"
rep_origin complement(5822..6410)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
CDS complement(6581..7441)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/note="This feature has 2 segments:
???1:?6581?..?7372?/?#ccffcc
???2:?7373?..?7441?/?#ccffcc?/?signal?sequence
Cleavage site after base 7372"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7442..7546)
/gene="bla"
/note="AmpR promoter"
/note="color: #ffffff; direction: LEFT"
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 tcgtggccgg catcaccggc gccacaggtg cggttgctgg
4381 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
4441 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
4501 ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg
4561 ctgcttccta atgcaggagt cgcataaggg agagcgtcga gtatctatga ttggaagtat
4621 gggaatggtg atacccgcat tcttcagtgt cttgaggtct cctatcagat tatgcccaac
4681 taaagcaacc ggaggaggag atttcatggt aaatttctct gacttttggt catcagtaga
4741 ctcgaactgt gagactatct cggttatgac agcagaaatg tccttcttgg agacagtaaa
4801 tgaagtccca ccaataaaga aatccttgtt atcaggaaca aacttcttgt ttcgaacttt
4861 ttcggtgcct tgaactataa aatgtagagt ggatatgtcg ggtaggaatg gagcgggcaa
4921 atgcttacct tctggacctt caagaggtat gtagggtttg tagatactga tgccaacttc
4981 agtgacaacg ttgctatttc gttcaaacca ttccgaatcc agagaaatca aagttgtttg
5041 tctactattg atccaagcca gtgcggtctt gaaactgaca atagtgtgct cgtgttttga
5101 ggtcatcttt gtatgaataa atctagtctt tgatctaaat aatcttgacg agccaaggcg
5161 ataaataccc aaatctaaaa ctcttttaaa acgttaaaag gacaagtatg tctgcctgta
5221 ttaaacccca aatcagctcg tagtctgatc ctcatcaact tgaggggcac tatcttgttt
5281 tagagaaatt tgcggagatg cgatatcgag aaaaaggtac gctgatttta aacgtgaaat
5341 ttatctcaag atctctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
5401 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
5461 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
5521 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
5581 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
5641 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
5701 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
5761 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
5821 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
5881 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
5941 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
6001 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
6061 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
6121 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
6181 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
6241 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
6301 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
6361 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
6421 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
6481 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
6541 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
6601 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
6661 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
6721 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
6781 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
6841 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
6901 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
6961 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
7021 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
7081 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
7141 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
7201 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
7261 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
7321 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
7381 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
7441 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
7501 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
7561 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
7621 gtatcacgag gccctttcgt cttcaagaat taattctcat gtttgacagc ttatcatcga
7681 taagctgact catgttggta ttgtgaaata gacgcagatc gggaacactg aaaaataaca
7741 gttattattc g
//
