pBridge

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  • Model: PVT4034
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pBridge

PVT4034    2ug
 

 

pBridge Information

Promoter: ADH1, TRP1, MET17

Replicator: 2 ori, ori

Plasmid classification: yeast series, yeast three hybrid carrier

Plasmid size: 6526bp

Prokaryotic resistance: Amp

Screening markers: TRP1

Cloned strain: DH5 alpha

Culture conditions: 37 centigrade, aerobic LB

Expression host: yeast cells

5'sequencing primers: M13R:CAGGAAACAGCTATGACC

3'sequencing primers: primers designed according to sequence

Use: Yeast expression

 

pBridge Description

pBridge is a shuttle vector that replicates autonomously in both E. coli and S. cerevisiae. It carries the bla gene (for ampicillin resistance in E. coli) and the TRP1 nutritional marker that allow yeast auxotrophs carrying pBridge to grow on limiting synthetic medium lacking tryptophan. Note: Yeast strain Y187 is a methionine auxotroph; therefore, haploid Y187 harboring pBridge cannot be grown on medium lacking methionine. 

pBridge

 

pBridge Sequence

LOCUS       Exported                6526 bp ds-DNA     circular SYN 10-SEP-2016

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    Untitled 18

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 6526)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4

FEATURES             Location/Qualifiers

     source          1..6526

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     rep_origin      1..1165

                     /note="2u ori"

                     /note="yeast 2u plasmid origin of replication"

     promoter        1447..1551

                     /gene="bla"

                     /note="AmpR promoter"

     CDS             1552..2412

                     /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"

     rep_origin      2583..3171

                     /direction=RIGHT

                     /note="ori"

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     terminator      3432..3702

                     /gene="S. cerevisae PGK1"

                     /note="PGK1 terminator"

                     /note="transcription terminator for 3-phosphoglycerate 

                     kinase gene"

     CDS             complement(3828..3848)

                     /codon_start=1

                     /product="nuclear localization signal of SV40 large T 

                     antigen"

                     /note="SV40 NLS"

                     /translation="PKKKRKV"

     CDS             complement(3870..3896)

                     /codon_start=1

                     /product="HA (human influenza hemagglutinin) epitope tag"

                     /note="HA"

                     /translation="YPYDVPDYA"

     promoter        complement(3918..4268)

                     /gene="S. cerevisiae MET17"

                     /gene_synonym="MET15; MET25"

                     /note="MET17 promoter"

                     /note="expression is repressed by methionine"

     promoter        4498..4528

                     /note="lac promoter"

                     /note="promoter for the E. coli lac operon"

     protein_bind    4536..4552

                     /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)."

     primer_bind     4560..4576

                     /note="M13 rev"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     promoter        4590..4691

                     /gene="S. cerevisiae TRP1"

                     /note="TRP1 promoter"

     CDS             4692..5366

                     /codon_start=1

                     /gene="S. cerevisiae TRP1"

                     /product="phosphoribosylanthranilate isomerase, required 

                     for tryptophan biosynthesis"

                     /note="TRP1"

                     /note="yeast auxotrophic marker"

                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK

                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES

                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW

                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA

                     KK"

     terminator      5419..5606

                     /gene="S. cerevisiae ADH1"

                     /note="ADH1 terminator"

                     /note="transcription terminator for the S. cerevisiae 

                     alcohol dehydrogenase 1 (ADH1) gene"

     CDS             complement(5653..6093)

                     /codon_start=1

                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"

                     /product="DNA binding domain of the GAL4 transcriptional 

                     activator"

                     /note="GAL4 DNA binding domain"

                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK

                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK

                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"

     promoter        complement(6121..6517)

                     /gene="S. cerevisiae ADH1"

                     /note="ADH1 promoter"

                     /note="promoter for alcohol dehydrogenase 1"

ORIGIN

        1 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt

       61 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt

      121 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa

      181 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc

      241 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc

      301 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg

      361 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg

      421 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta

      481 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat

      541 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag

      601 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca

      661 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa

      721 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg

      781 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga

      841 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc

      901 cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc

      961 tctgaagttc ctatactttc tagctagaga ataggaactt cggaatagga acttcaaagc

     1021 gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac agctcactgt

     1081 tcacgtcgca cctatatctg cgtgttgcct gtatatatat atacatgaga agaacggcat

     1141 agtgcgtgtt tatgcttaaa tgcgttatgg tgcactctca gtacaatctg ctctgatgcc

     1201 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt

     1261 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag

     1321 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt

     1381 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga

     1441 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc

     1501 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt

     1561 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct

     1621 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt

     1681 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt

     1741 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac

     1801 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac

     1861 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct

     1921 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg

     1981 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg

     2041 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca

     2101 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa

     2161 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt

     2221 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc

     2281 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg

     2341 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt

     2401 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt

     2461 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc

     2521 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct

     2581 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta

     2641 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc

     2701 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac

     2761 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct

     2821 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat

     2881 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg

     2941 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa

     3001 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg

     3061 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga

     3121 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc

     3181 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct

     3241 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct

     3301 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca

     3361 atacgcaaac gcctctcccc gcgcgttggc cgattcatta atgcaggaag atccgaggcc

     3421 tagctttaac gaacgcagaa ttttcgagtt attaaactta aaatacgctg aacccgaaca

     3481 tagaaatatc gaatgggaaa aaaaaactgc ataaaggcat taaaagagga gcgaattttt

     3541 ttttaataaa aatcttaata atcattaaaa gataaataat agtctatata tacgtatata

     3601 aataaaaaat attcaaaaaa taaaataaac tattatttta gcgtaaagga tggggaaaga

     3661 gaaaagaaaa aaattgatct atcgatttca attcaattca atttatttct tttcggataa

     3721 gaaagcaaca cctggcaatt ccttaccttc caataattcc aaagaagcac caccaccagt

     3781 agagacatgg gagatcagcc cgaagatctt cgggctaatg cggccgccac ctttctcttc

     3841 ttcttaggac tagaaggtcc tcccaggctg gcatagtcag gcacgtcata aggatagcta

     3901 gaagccatat ggcccattgt atggatgggg gtaatagaat tgtatctatg tatctgacga

     3961 ccctgtatta cacgaaggaa atagtagatg aaaagacaag agagcaagaa aaaggaaaaa

     4021 cttcctttct aacagacgct ttacttaacc ttatatatat cttatttttt tcatcgagcg

     4081 tgttcaattg gacaaggtgc caccttttcg acacttcggt tattatgtta cacagttttc

     4141 atgaggatgg cgccttgact aacttaatta gtcatttgcc aaccaccaca gttccccaat

     4201 atcgacagct tcacgtgcca tttgccattt tgcgatcatg tgacctcaag agaaaaagca

     4261 aaaaataatg agagctaagg gattcgaacc cttgcatccg aagatatcag agattttaga

     4321 ggttaaatcc acctaaatct gacgccttaa accactcggc caaactccca ttgtatgttg

     4381 gaagaattag aagctaggcc tcggatcttc ctggcacgac aggtttcccg actggaaagc

     4441 gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact cattaggcac cccaggcttt

     4501 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac

     4561 aggaaacagc tatgacatga ttacgaatta attcggtcga aaaaagaaaa ggagagggcc

     4621 aagagggagg gcattggtga ctattgagca cgtgagtata cgtgattaag cacacaaagg

     4681 cagcttggag tatgtctgtt attaatttca caggtagttc tggtccattg gtgaaagttt

     4741 gcggcttgca gagcacagag gccgcagaat gtgctctaga ttccgatgct gacttgctgg

     4801 gtattatatg tgtgcccaat agaaagagaa caattgaccc ggttattgca aggaaaattt

     4861 caagtcttgt aaaagcatat aaaaatagtt caggcactcc gaaatacttg gttggcgtgt

     4921 ttcgtaatca acctaaggag gatgttttgg ctctggtcaa tgattacggc attgatatcg

     4981 tccaactgca tggagatgag tcgtggcaag aataccaaga gttcctcggt ttgccagtta

     5041 ttaaaagact cgtatttcca aaagactgca acatactact cagtgcagct tcacagaaac

     5101 ctcattcgtt tattcccttg tttgattcag aagcaggtgg gacaggtgaa cttttggatt

     5161 ggaactcgat ttctgactgg gttggaaggc aagagagccc cgaaagctta cattttatgt

     5221 tagctggtgg actgacgcca gaaaatgttg gtgatgcgct tagattaaat ggcgttattg

     5281 gtgttgatgt aagcggaggt gtggagacaa atggtgtaaa agactctaac aaaatagcaa

     5341 atttcgtcaa aaatgctaag aaataggtta ttactgagta gtatttattt aagtattgtt

     5401 tgtgcacttg ccggcatgcc ggtagaggtg tggtcaataa gagcgacctc atgctatacc

     5461 tgagaaagca acctgaccta caggaaagag ttactcaaga acaagaattt tcgttttaaa

     5521 acctaagagt cactttaaaa tttgtataca cttatttttt ttataactta tttaataata

     5581 aaaatcataa atcataagaa attcgcccgg aattagcttg gctgcaggtc gacggatccc

     5641 cgggaattcc ggcgatacag tcaactgtct ttgacctttg ttactactct cttccgatga

     5701 tgatgtcgca cttattctat gctgtctcaa tgttagaggc atatcagtct ccactgaagc

     5761 caatctatct gtgacggcat ctttattcac attatcttgt acaaataatc ctgttaacaa

     5821 tgcttttata tcctgtaaag aatccatttt caaaatcatg tcaaggtctt ctcgaggaaa

     5881 aatcagtaga aatagctgtt ccagtctttc tagccttgat tccacttctg tcagatgtgc

     5941 cctagtcagc ggagaccttt tggttttggg agagtagcga cactcccagt tgttcttcag

     6001 acacttggcg cacttcggtt tttctttgga gcacttgagc tttttaagtc ggcaaatatc

     6061 gcatgcttgt tcgatagaag acagtagctt catctttcag gaggcttgct tcaagcttgg

     6121 agttgattgt atgcttggta tagcttgaaa tattgtgcag aaaaagaaac aaggaagaaa

     6181 gggaacgaga acaatgacga ggaaacaaaa gattaataat tgcaggtcta tttatacttg

     6241 atagcaagac agcaaacttt tttttatttc aaattcaagt aactggaagg aaggccgtat

     6301 accgttgctc attagagagt agtgtgcgtg aatgaaggaa ggaaaaagtt tcgtgtgctt

     6361 cgagataccc ctcatcagct ctggaacaac gacatctgtt ggtgctgtct ttgtcgttaa

     6421 ttttttcctt tagtgtcttc catcattttt ttgtcattgc ggatatggtg agacaacaac

     6481 gggggagaga gaaaagaaaa aaaaagaaaa gaagttgcat gcaagc

//

 

 

Caution:
1.  This product is FOR RESEARCH USE ONLY!
2.  The item is lyophilized form, Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.

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