pPINK- LC Plasmid

$216.00

  • Model: PVT4035
  • 49 Units in Stock
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pPINK-LC

PVT4035    2ug

 

pPINK-LC Information

 

Promoter: PpADE2 LC, AOX1

Replicon: pUC ori

Terminator: CYC1

Plasmid Classification: Yeast Series, Pichia pastoris Expression Vector

Plasmid size: 7732 BP

Prokaryotic resistance: Amp

Screening marker: ADE2

Cloning strain: DH5alpha

Culture conditions: 37 C, aerobic LB

Expression host: yeast cells

5'Sequencing Primers: AOX5: GACTGTTCCAATTGACAAGC

3'Sequencing primers: CYC1-R: GCGTGAATGTAA GCGTGAC

 

pPINK-LC cloning site

pPINK-LC-vector

 

pPINK-LC Sequence

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

//

 

Caution:
Product is for research use only!

 

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pPINK-LC,Plasmid pPINK-LC,pPINK-LC vector

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