基本信息
| 启动子: | PpADE2 LC，AOX1 |
| 复制子: | pUC ori |
| 终止子: | CYC1 |
| 质粒分类: | 酵母系列，毕赤酵母表达载体 |
| 质粒大小: | 7732bp |
| 原核抗性: | Amp |
| 筛选标记: | ADE2 |
| 克隆菌株: | DH5α |
| 培养条件: | 37℃，有氧 LB |
| 表达宿主: | 酵母细胞 |
| 5'测序引物: | AOX5:GACTGGTTCCAATTGACAAGC |
| 3'测序引物: | CYC1-R: GCGTGAATGTAAGCGTGAC |
质粒简介
he PichiaPink™ Vector Kit contains the pPINK-HC and pPINK-LC vectors for use with the PichiaPink™ Yeast Expression System. The pPINK vectors are built around the ADE2 gene for complementing the ade2 deletion in the PichiaPink™ strains. You can order the PichiaPink™ Vector Kit to refill your PichiaPink™ Yeast Expression System Kits.
质粒图谱
质粒序列
LOCUS Exported 7732 bp ds-DNA circular SYN 10-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 15
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7732)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..7732
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 2..940
/gene="Pichia pastoris AOX1"
/note="AOX1 promoter"
/note="inducible promoter, regulated by methanol"
terminator 1013..1262
/gene="S. cerevisiae CYC1"
/note="CYC1 terminator"
/note="transcription terminator for CYC1"
promoter 1282..1363
/gene="Pichia pastoris ADE2"
/note="PpADE2 LC promoter"
/note="strong 82-bp promoter fragment that requires only
low-copy integration for efficient expression"
CDS 1364..3055
/codon_start=1
/gene="Pichia pastoris ADE2"
/product="phosphoribosylaminoimidazole carboxylase from
Pichia pastoris"
/note="PpADE2"
/note="essential for the biosynthesis of purine
nucleotides, including adenine"
/translation="MDSQVIGILGGGQLGRMIVEAASRLNIKTVILDDGFSPAKHINAA
QDHIDGSFKDEEAIAKLAAKCDVLTVEIEHVNTDALKRVQDRTGIKIYPLPETIELIKD
KYLQKEHLIKHNISVTKSQGIESNEKALLLFGEENGFPYLLKSRTMAYDGRGNFVVESK
EDISKALEFLKDRPLYAEKFAPFVKELAVMVVRSLEGEVFSYPTVETVHKDNICHIVYA
PARVNDTIQKKAQILAENTVKTFPGAGIFGVEMFLLSDGELLVNEIAPRPHNSGHYTID
ACVTSQFEAHVRAITGLPMPLDFTKLSTSNTNAIMLNVLGAEKSHGELEFCRRALETPG
ASVYLYGKTTRLARKMGHINIIGSSMLEAEQKLEYILEESTHLPSSTVSADTKPLVGVI
MGSDSDLPVISKGCDILKQFGVPFEVTIVSAHRTPQRMTRYAFEAASRGIKAIIAGAGG
AAHLPGMVAAMTPLPVIGVPVKGSTLDGVDSLHSIVQMPRGVPVATVAINNATNAALLA
IRILGTIDHKWQKEMSKYMNAMETEVLGKASNLESEGYESYLKNRL"
promoter 3282..3431
/gene="Pichia pastoris TRP2"
/note="PpTRP2 promoter"
/note="promoter for Pichia pastoris TRP2"
CDS 3432..5048
/codon_start=1
/gene="Pichia pastoris TRP2"
/product="anthranilate synthase from Pichia pastoris"
/note="PpTRP2"
/note="catalyzes the initial step of tryptophan
biosynthesis"
/translation="MSVSDSHLATDYFGTQLTKQIHPSAESFLDIVKNDRQSHSPETVP
NLYPIYKFIQSDFLTPHLAYIKLTNSSETPSPSFLLESAKNGDTVDRYTFMGHSPRKII
KTGPLEGAEVDPLVLLEKELKGTRQAQLPGIPRLSGGAIGYISYDCIKYFEPKTERKLK
DVLQLPEAALMLFDTIVAFDNVYQRFQVIGNVSLSVDDSDEAILEKYYKTREEVEKISK
VVFDNKTVPYYEQKDIIQGQTFTSNIGQEGYENHVRKLKEHILKGDIFQAVPSQRVARP
TSLHPFNIYRHLRTVNPSPYMFYIDYLDFQVVGASPELLVKSDNNNKIITHPIAGTLPR
GKTIEEDDNYAKQLKSSLKDRAEHVMLVDLARNDINRVCEPTSTTVDRLLTVERFSHVM
HLVSEVSGTLRPNKTRFDAFRSIFPAGTVSGAPKVRAMQLIGELEGEKRGVYAGAVGHW
SYDGKSMDTCIALRTMVVKDGVAYLQAGGGIVYDSDPYDEYIETMNKMRSNNNTILEAE
KIWTDRLARDENQSESEENDQ"
primer_bind complement(5110..5126)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 5134..5150
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(5158..5188)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 5203..5224
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5512..6100)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6271..7131)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7132..7236)
/gene="bla"
/note="AmpR promoter"
primer_bind 7710..7726
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
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 aaagtcggca 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 attcgaaacg gaattccgga ccggcatgca
961 ggcctggtac cggccggcca tttaaataca ggcccctttt cctttgtcga tatcatgtaa
1021 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
1081 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
1141 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaacgc gtgtacgcat
1201 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt
1261 gcaagctgga tccgcggccg cgacttgact cttcactagc ctatgcaaat aaggttacct
1321 tttccaagaa tcgtagaaac gattaaaaaa cttccaaact ctcatggatt ctcaggtaat
1381 aggtattcta ggaggaggcc agctaggccg aatgattgtt gaggccgcta gcaggctcaa
1441 tatcaagacc gtgattcttg atgatggttt ttcacctgct aagcacatta atgctgcgca
1501 agaccacatc gacggatcat tcaaagatga ggaggctatc gccaagttag ctgccaaatg
1561 tgatgttctc actgtagaga ttgagcatgt caacacagat gctctaaaga gagttcaaga
1621 cagaactgga atcaagatat atcctttacc agagacaatc gaactaatca aggataagta
1681 cttgcaaaag gaacatttga tcaagcacaa catttcggtg acaaagtctc agggtataga
1741 atctaatgaa aaggcgctgc ttttgtttgg agaagagaat ggatttccat atctgttgaa
1801 gtcccggact atggcttatg atggaagagg caattttgta gtggagtcta aagaggacat
1861 cagtaaggca ttagagttct tgaaagatcg tccattgtat gccgagaagt ttgctccttt
1921 tgttaaagaa ttagcggtaa tggttgtgag atcactggaa ggcgaagtat tctcctaccc
1981 aaccgtagaa actgtgcaca aggacaatat ctgtcatatt gtgtatgctc cggccagagt
2041 taatgacacc atccaaaaga aagctcaaat attagctgaa aacactgtga agactttccc
2101 aggcgctgga atcttcggag ttgagatgtt cctattgtct gatggagaac ttcttgtaaa
2161 tgagattgct ccaaggcccc acaattctgg tcactataca atcgatgcat gtgtaacatc
2221 tcagttcgaa gcacatgtaa gagccataac tggtctgcca atgccactag atttcaccaa
2281 actatctact tccaacacca acgctattat gctcaatgtt ttgggtgctg aaaaatctca
2341 cggggaatta gagttttgta gaagagcctt agaaacaccc ggtgcttctg tatatctgta
2401 cggaaagacc acccgattgg ctcgtaagat gggtcatatc aacataatag gatcttccat
2461 gttggaagca gaacaaaagt tagagtacat tctagaagaa tcaacccact taccatccag
2521 tactgtatca gctgacacta aaccgttggt tggagttatc atgggttcag actctgatct
2581 acctgtgatt tcgaaaggtt gcgatatttt aaaacagttt ggtgttccat tcgaagttac
2641 tattgtctct gctcatagaa caccacagag aatgaccaga tatgcctttg aagccgctag
2701 tagaggtatc aaggctatca ttgcaggtgc tggtggtgct gctcatcttc caggaatggt
2761 tgctgccatg actccgttgc cagtcattgg tgttcctgtc aagggctcta cgttggatgg
2821 tgtagactcg ctacactcga ttgtccaaat gcctagaggt gttcctgtgg ctacggttgc
2881 tatcaacaac gccaccaatg ccgctctgtt ggccatcagg attttaggta caattgacca
2941 caaatggcaa aaggaaatgt ccaagtatat gaatgcaatg gagaccgaag tgttggggaa
3001 ggcatccaac ttggaatctg aagggtatga atcctatttg aagaatcgtc tttgaattta
3061 gtattgtttt ttaatagatg tatatataat agtacacgta acttatctat tccattcata
3121 attttatttt aaaggttcgg tagaaatttg tcctccaaaa agttggttag agcctggcag
3181 ttttgatagg cattattata gattgggtaa tatttaccct gcacctggag gaactttgca
3241 aagagcctca tgtgcggcgc gccaggccat aatggccaaa cggtttctca attactatat
3301 actactaacc atttacctgt agcgtatttc ttttccctct tcgcgaaagc tcaagggcat
3361 cttcttgact catgaaaaat atctggattt cttctgacag atcatcaccc ttgagcccaa
3421 ctctctagcc tatgagtgta agtgatagtc atcttgcaac agattatttt ggaacgcaac
3481 taacaaagca gatacaccct tcagcagaat cctttctgga tattgtgaag aatgatcgcc
3541 aaagtcacag tcctgagaca gttcctaatc tttaccccat ttacaagttc atccaatcag
3601 acttcttaac gcctcatctg gcttatatca agcttaccaa cagttcagaa actcccagtc
3661 caagtttctt gcttgaaagt gcgaagaatg gtgacaccgt tgacaggtac acctttatgg
3721 gacattcccc cagaaaaata atcaagactg ggcctttaga gggtgctgaa gttgacccct
3781 tggtgcttct ggaaaaagaa ctgaagggca ccagacaagc gcaacttcct ggtattcctc
3841 gtctaagtgg tggtgccata ggatacatct cgtacgattg tattaagtac tttgaaccaa
3901 aaactgaaag aaaactgaaa gatgttttgc aacttccgga agcagctttg atgttgttcg
3961 acacgatcgt ggcttttgac aatgtttatc aaagattcca ggtaattgga aacgtttctc
4021 tatccgttga tgactcggac gaagctattc ttgagaaata ttataagaca agagaagaag
4081 tggaaaagat cagtaaagtg gtatttgaca ataaaactgt tccctactat gaacagaaag
4141 atattattca aggccaaacg ttcacctcta atattggtca ggaagggtat gaaaaccatg
4201 ttcgcaagct gaaagaacat attctgaaag gagacatctt ccaagctgtt ccctctcaaa
4261 gggtagccag gccgacctca ttgcaccctt tcaacatcta tcgtcatttg agaactgtca
4321 atccttctcc atacatgttc tatattgact atctagactt ccaagttgtt ggtgcttcac
4381 ctgaattact agttaaatcc gacaacaaca acaaaatcat cacacatcct attgctggaa
4441 ctcttcccag aggtaaaact atcgaagagg acgacaatta tgctaagcaa ttgaagtcgt
4501 ctttgaaaga cagggccgag cacgtcatgc tggtagattt ggccagaaat gatattaacc
4561 gtgtgtgtga gcccaccagt accacggttg atcgtttatt gactgtggag agattttctc
4621 atgtgatgca tcttgtgtca gaagtcagtg gaacattgag accaaacaag actcgcttcg
4681 atgctttcag atccattttc ccagcaggaa ccgtctccgg tgctccgaag gtaagagcaa
4741 tgcaactcat aggagaattg gaaggagaaa agagaggtgt ttatgcgggg gccgtaggac
4801 actggtcgta cgatggaaaa tcgatggaca catgtattgc cttaagaaca atggtcgtca
4861 aggacggtgt cgcttacctt caagccggag gtggaattgt ctacgattct gacccctatg
4921 acgagtacat cgaaaccatg aacaaaatga gatccaacaa taacaccatc ttggaggctg
4981 agaaaatctg gaccgatagg ttggccagag acgagaatca aagtgaatcc gaagaaaacg
5041 atcaatgaac ggaggacgta agtaggaatt tatggtttgg ccataatggc ctagcttggc
5101 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
5161 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
5221 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
5281 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
5341 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
5401 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
5461 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
5521 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
5581 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
5641 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
5701 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
5761 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
5821 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
5881 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
5941 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
6001 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
6061 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
6121 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
6181 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
6241 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
6301 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
6361 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
6421 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
6481 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
6541 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
6601 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
6661 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
6721 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
6781 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
6841 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
6901 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
6961 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
7021 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
7081 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
7141 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
7201 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
7261 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
7321 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
7381 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
7441 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
7501 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
7561 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
7621 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
7681 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tg
温馨提示：不可用于临床ZL。
