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基本信息

質粒圖譜和多克隆位點信息

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載體描述

帶有HIS3標記的酵母表達載體，用半乳糖調節表達和標記2個基因。

載體序列

LOCUS       Exported                6705 bp ds-DNA     circular SYN 02-APR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pESC-His

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 6705)

  AUTHORS   BT-Lab（Agilent）

  TITLE     Direct Submission

  JOURNAL   Exported Tuesday, Apr 2, 2019 from SnapGene Viewer 4.3.6

FEATURES             Location/Qualifiers

     source          1..6705

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        317..503

                     /gene="S. cerevisiae HIS3"

                     /label=HIS3 promoter

     CDS             504..1163

                     /codon_start=1

                     /gene="S. cerevisiae HIS3"

                     /product="imidazoleglycerol-phosphate dehydratase, required

                     for histidine biosynthesis"

                     /label=HIS3

                     /note="yeast auxotrophic marker"

                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE

                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF

                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL

                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"

     rep_origin      complement(1423..1878)

                     /direction=LEFT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow

                     indicates direction of (+) strand synthesis"

     terminator      1961..2126

                     /gene="S. cerevisiae ADH1"

                     /label=ADH1 terminator

                     /note="transcription terminator for the S. cerevisiae

                     alcohol dehydrogenase 1 (ADH1) gene"

     CDS             complement(2274..2297)

                     /codon_start=1

                     /product="FLAG(R) epitope tag, followed by an enterokinase

                     cleavage site"

                     /label=FLAG

                     /translation="DYKDDDDK"

     misc_feature    2299..2339

                     /label=MCS1

     promoter        2344..3008

                     /gene="GAL1 and GAL10 genes of S. cerevisiae"

                     /label=GAL1,10 promoter

                     /note="divergent inducible promoter, regulated by Gal4"

     protein_bind    2560..2677

                     /label=UAS

                     /bound_moiety="Gal4"

                     /note="upstream activating sequence mediating

                     Gal4-dependent induction"

     promoter        3019..3037

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     misc_feature    3038..3051

                     /label=MCS2

                     /note="multiple cloning site"

     CDS             3055..3084

                     /codon_start=1

                     /product="Myc (human c-Myc proto-oncogene) epitope tag"

                     /label=Myc

                     /translation="EQKLISEEDL"

     terminator      3114..3303

                     /gene="S. cerevisiae CYC1"

                     /label=CYC1 terminator

                     /note="transcription terminator for CYC1"

     rep_origin      complement(3546..4134)

                     /direction=LEFT

                     /label=ori

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

                     replication"

     CDS             complement(4305..5165)

                     /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"

     promoter        complement(5166..5270)

                     /gene="bla"

                     /label=AmpR promoter

     rep_origin      5297..6639

                     /label=2u ori

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

     protein_bind    complement(6260..6307)

                     /label=FRT

                     /bound_moiety="FLP recombinase from the Saccharomyces

                     cerevisiae 2u plasmid"

                     /note="FLP-mediated recombination occurs in the 8-bp core

                     sequence TCTAGAAA (Turan and Bode, 2011)."

ORIGIN

        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca

       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg

      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc

      181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt

      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta

      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat

      361 tttttttttt cccctagcgg atgactcttt ttttttctta gcgattggca ttatcacata

      421 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc

      481 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa

      541 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact

      601 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga

      661 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt

      721 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca

      781 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctactg gcgcgtggag

      841 taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga gcggtggtag

      901 atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag aaagtaggag

      961 atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct agcagaatta

     1021 ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag agtgcgttca

     1081 aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac gatgttccct

     1141 ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca tacgatatat

     1201 atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat

     1261 actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt

     1321 ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc tatgcggtgt

     1381 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata

     1441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg

     1501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc

     1561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa

     1621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt

     1681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac

     1741 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta

     1801 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg

     1861 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc

     1921 gatcggtgcg ggcctcttcg ctattacgcc agctgaattg gagcgacctc atgctatacc

     1981 tgagaaagca acctgaccta caggaaagag ttactcaaga ataagaattt tcgttttaaa

     2041 acctaagagt cactttaaaa tttgtataca cttatttttt ttataactta tttaataata

     2101 aaaatcataa atcataagaa attcgcttat ttagaagtgt caacaacgta tctaccaacg

     2161 atttgaccct tttccatctt ttcgtaaatt tctggcaagg tagacaagcc gacaaccttg

     2221 attggagact tgaccaaacc tctggcgaag aattgttaat taagagctca gatcttatcg

     2281 tcgtcatcct tgtaatccat cgatactagt gcggccgccc tttagtgagg gttgaattcg

     2341 aattttcaaa aattcttact ttttttttgg atggacgcaa agaagtttaa taatcatatt

     2401 acatggcatt accaccatat acatatccat atacatatcc atatctaatc ttacttatat

     2461 gttgtggaaa tgtaaagagc cccattatct tagcctaaaa aaaccttctc tttggaactt

     2521 tcagtaatac gcttaactgc tcattgctat attgaagtac ggattagaag ccgccgagcg

     2581 ggtgacagcc ctccgaagga agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt

     2641 cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt ctacaatact

     2701 agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa accttcaaat

     2761 gaacgaatca aattaacaac cataggatga taatgcgatt agttttttag ccttatttct

     2821 ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat aaatgcaaaa

     2881 actgcataac cactttaact aatactttca acattttcgg tttgtattac ttcttattca

     2941 aatgtaataa aagtatcaac aaaaaattgt taatatacct ctatacttta acgtcaagga

     3001 gaaaaaaccc cggatccgta atacgactca ctatagggcc cgggcgtcga catggaacag

     3061 aagttgattt ccgaagaaga cctcgagtaa gcttggtacc gcggctagct aagatccgct

     3121 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata

     3181 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga

     3241 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg

     3301 aagatccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg

     3361 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg

     3421 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga

     3481 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg

     3541 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag

     3601 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc

     3661 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg

     3721 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt

     3781 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc

     3841 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc

     3901 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg

     3961 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca

     4021 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc

     4081 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat

     4141 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt

     4201 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt

     4261 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc

     4321 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc

     4381 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata

     4441 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg

     4501 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc

     4561 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct

     4621 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa

     4681 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt

     4741 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca

     4801 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac

     4861 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca

     4921 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt

     4981 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc

     5041 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca

     5101 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata

     5161 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc

     5221 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc

     5281 cgaaaagtgc cacctgaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc

     5341 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa

     5401 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc

     5461 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa

     5521 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa

     5581 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt

     5641 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag

     5701 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg

     5761 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat

     5821 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg

     5881 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt

     5941 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac

     6001 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt

     6061 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag

     6121 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta

     6181 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt

     6241 tggttttcaa aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag

     6301 gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat

     6361 acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga

     6421 gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta

     6481 ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat

     6541 gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt

     6601 ctcatccttc aatgctatca tttcctttga tattggatca tctaagaaac cattattatc

     6661 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgt

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