pTEF:ATP pTEF- ATP

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  • Model: PVT11292
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pTEF:ATP pTEF-ATP

PVT11292    2ug

 

pTEF:ATP pTEF-ATP Information

Promoter: TEF1

Replicon: pUC

Prokaryotic resistance: ampicillin Amp

Screening marker: hereditary mycin G418

Clone strain: Escherichia coli DH5 alpha

Culture conditions: 37 C

 

pTEF:ATP pTEF-ATP Description

PTEF:ATP pTEF-ATP is a yeast ATP sensor plasmid. HO-TEF1p-ATeam1.03-KanMX4-HO. Codon optimized for yeast version of the ATeam1.03 ATP FRET sensor (originally from Imamura et al. PNAS, 2009 ), expressed via the TEF1 promoter. HO locus integration.

 

pTEF:ATP pTEF-ATP Sequence

 

LOCUS       Exported                8554 bp ds-DNA     circular SYN 26-DEC-2017
DEFINITION  synthetic circular DNA
FEATURES             Location/Qualifiers
     source          1..8554
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    281..1187
                     /label=HO up
     terminator      1205..1394
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(1427..3262)
                     /codon_start=1
                     /label=ATeam1.03
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAIDMKTVKVNITTPDGPVYDADIEMVSVRAESGDLGILPGHIPTKAPLKIGAVR
                     LKKDGQTEMVAVSGGTVEVRPDHVTINAQAAETAEGIDKERAEAARQRAQERLNSQSDD
                     TDIRRAELALQRALNRLDVAGKANEFMDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQ
                     SKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELD
                     GDVNGHKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHM
                     KQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNIL
                     GHKLEYNYNSHNVYITADKQKNGIKANFKIRHNIE"
     promoter        complement(3280..3678)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3761..4104
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             4105..4914
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      4920..5117
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    5178..5675
                     /label=HO down
     promoter        6639..6743
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6744..7604
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7775..8363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
       61 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      121 ggcaccccag gctttacact ttatgcttcc gcggctcgta tgttgtgtgg aattgtgagc
      181 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccactag tccgaggcct
      241 cgagatccga tatcgccgtg gcggccgcca gctgaagctt aattatcctg ggcacgagtg
      301 aaacaaagct aaaaccttta tttagcatgg ccattgaatg taacaattat atatatcgca
      361 agcacaaaaa atcaaggaga gagaactacc actttgttca tgtgtacaat gttcattatc
      421 tccataagca aaaaaaaaaa aatagaaaac atatgctata aggttgatat tctcacgagt
      481 aagcggcact tgctacttat tgacattgca gatttttggc tacagaaata gtatattaga
      541 gattataatt gctaatcaaa tcaaaatata aaattagtaa accaaaccat ttataccctt
      601 ccttagtagt tatggattgt tttttaatga tatttctgca aaccaaagaa agattgttat
      661 ccagatagaa tttagttttg atattcattt ttttgttgaa gattgaacgc catatctggg
      721 cctcataatt caaaagacgg tgccattatc ggtagcgttt cgcattgtac tggatttcag
      781 aaatttcaca gttgatgaat cgaaaagaat ggtctcattg caacacgtaa ggttaagatg
      841 tcccttttta ccattatagg caataaatga atcataaaac gaccgtatac tggtgaaata
      901 gtagggagaa cgagtacctg tagtaaaaag tataaatcat agttaatcgg gcaatgtccc
      961 tcgatcaagg agtattgtgt catgttcgag acaaacgcca acatttttgt ttcttttgga
     1021 caaatgttgt ttgcatttat gatccgttat attttgatct aatgtagagt tgcacgtagt
     1081 tcttactggc aaagaaatcg atgcatacca aaaaagaata aaggtgatat ttgatcttta
     1141 ccgtttagtt ccaacgtaaa attgtgcctt tggacttaaa atggcgtcgt acgctgagct
     1201 ggatcttcga gcgtcccaaa accttctcaa gcaaggtttt cagtataatg ttacatgcgt
     1261 acacgcgtct gtacagaaaa aaaagaaaaa tttgaaatat aaataacgtt cttaatacta
     1321 acataactat aaaaaaataa atagggacct agacttcagg ttgtctaact ccttcctttt
     1381 cggttagagc ggatcttagc tagccgcggt accaagctta ctcgagtcat tcaatgttat
     1441 gtctaatttt gaagttggcc ttgataccat ttttttgttt gtcagcagtg atgtagacgt
     1501 tgtgagagtt ataattgtat tctaacttat gacctaaaat gttaccatct tctttaaaat
     1561 caataccttt caattcaatt ctattgacca aagtgtcacc ttcaaatttt acttcagctc
     1621 tggtcttgta attaccatcg tccttaaaga aaatagttct ttcttggaca taaccttcag
     1681 gcattgcaga cttaaaaaag tcatgttgct tcatgtggtc aggatatcta gcgaagcatt
     1741 gtaaaccgta acccaatgtt gtaactaatg taggccaagg aacaggtaac ttaccggtag
     1801 tgcatatcaa tttcaaagtc aatttaccgt atgttgcgtc accttcacct tcaccactaa
     1861 ctgaaaactt atgaccattg acgtcaccgt ctaattctac taaaataggg acgacacctg
     1921 tgaataattc ttcaccttta ctaaccatac caccagaacc acccttgtat aattcatcca
     1981 tacccaaagt aataccggca gcggtgacaa attctaataa aaccatatga tctctttttt
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