PCT74 -2 ug

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    pCT74

    Catalog No. PVT11337
    Packing 2ug

     

    pCT74 Informaiton

    Alias: pCT74-Hygro-sGFP; pCT-74

    Prokaryotic resistance: Ampicillin (Amp)

    Screening marker: hygromycin Hyg, green fluorescent protein EGFP

    Cloned strain: Escherichia coli DH5 alpha

    Culture conditions: 37℃

     

    pCT74 Description

    PCT74 is a fungal expression plasmid which can be transformed into various fungi by protoplast method.

     

    pCT74 Reference

    1) Clathrin mediated endocytosis is involved in the uptake of exogenous double-stranded RNA in the white mold phytopathogen Sclerotinia sclerotiorum. Sci Rep 10, 12773 (2020). 
    Author: Wytinck, N., Sullivan, D.S., Biggar, K.T. et al.
    Scientific Reports

    Received
    20 April 2020

    Accepted
    16 July 2020

    Published
    29 July 2020

    DOI
    https://doi.org/10.1038/s41598-020-69771-9

     

    pCT74 Sequence

    LOCUS       Exported                5790 bp ds-DNA     circular SYN 25-JAN-2018

    DEFINITION  synthetic circular DNA

    FEATURES             Location/Qualifiers

         source          1..5790

                         /organism="synthetic DNA construct"

                         /mol_type="other DNA"

         primer_bind     19..35

                         /label=SK primer

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

                         variants"

         terminator      78..330

                         /label=NOS terminator

                         /note="nopaline synthase terminator and poly(A) signal"

         CDS             complement(349..1068)

                         /codon_start=1

                         /product="enhanced GFP"

                         /label=EGFP

                         /note="mammalian codon-optimized"

                         /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL

                         KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD

                         GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK

                         VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL

                         EFVTAAGITHGMDELYK"

         CDS             1855..2880

                         /codon_start=1

                         /gene="aph(4)-Ia"

                         /product="aminoglycoside phosphotransferase from E. coli"

                         /label=HygR

                         /note="confers resistance to hygromycin"

                         /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG

                         YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP

                         ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ

                         TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF

                         GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG

                         NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK

                         E"

         promoter        complement(2947..2965)

                         /label=T3 promoter

                         /note="promoter for bacteriophage T3 RNA polymerase"

         primer_bind     complement(2986..3002)

                         /label=M13 rev

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

                         variants"

         protein_bind    3010..3026

                         /label=lac operator

                         /bound_moiety="lac repressor encoded by lacI"

                         /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(3034..3064)

                         /label=lac promoter

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

         protein_bind    3079..3100

                         /label=CAP binding site

                         /bound_moiety="E. coli catabolite activator protein"

                         /note="CAP binding activates transcription in the presence 

                         of cAMP."

         rep_origin      complement(3388..3976)

                         /direction=LEFT

                         /label=ori

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

                         replication"

         CDS             complement(4147..5007)

                         /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(5008..5112)

                         /gene="bla"

                         /label=AmpR promoter

         rep_origin      complement(5138..5593)

                         /direction=LEFT

                         /label=f1 ori

                         /note="f1 bacteriophage origin of replication; arrow 

                         indicates direction of (+) strand synthesis"

         primer_bind     5735..5751

                         /label=M13 fwd

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

                         variants"

         promoter        5758..5776

                         /label=T7 promoter

                         /note="promoter for bacteriophage T7 RNA polymerase"

    ORIGIN

            1 caccgcggtg gcggccgctc tagaactagt ggatcccccg ggctgcagga attctcatgt

           61 ttgacagctt atcatcggat ctagtaacat agatgacacc gcgcgcgata atttatccta

          121 gtttgcgcgc tatattttgt tttctatcgc gtattaaatg tataattgcg ggactctaat

          181 cataaaaacc catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac

          241 gtaattcaac agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa

          301 gaaactttat tgccaaatgt ttgaacgatc tgcagccggg cggccgcttt acttgtacag

          361 ctcgtccatg ccgtgagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc

          421 gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg

          481 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac

          541 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt

          601 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat

          661 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc

          721 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg

          781 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc

          841 ggggtagcgg ctgaagcact gcacgccgta ggtgaaggtg gtcacgaggg tgggccaggg

          901 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc

          961 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac

         1021 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg cctatattca

         1081 ttcattgtca gctatcgctg gcaacaagat ataagcgtgt acttacgatt ctaggaccct

         1141 gcgaatagag ttggagttgg ataccgatgg acgattgtta gataggacgg gatgctgata

         1201 gtcgaggtaa gttgttgagg gtggtggttt tatcaaggac ggacacctta gccctttata

         1261 tacacagaat cttattgcta ggtagccgga cggtgcggat tccaggtatc agtaggccct

         1321 ctgaatgtcc atgcaatgat gcccaacaat cgagacgatc ctacttcgga accgactcga

         1381 tcctcgtcgg gcctacatgt atgagaccat gtgcacctcc gaagcatcct tacggagagc

         1441 ggaattgcgc gtacagaact cctccatgca ttccaatcga taccgtcgac gttaactgat

         1501 attgaaggag cattttttgg gcttggctgg agctagtgga ggtcaacaat gaatgcctat

         1561 tttggtttag tcgtccaggc ggtgagcaca aaatttgtgt cgtttgacaa gatggttcat

         1621 ttaggcaact ggtcagatca gccccacttg tagcagtagc ggcggcgctc gaagtgtgac

         1681 tcttattagc agacaggaac gaggacatta ttatcatctg ctgcttggtg cacgataact

         1741 tggtgcgttt gtcaagcaag gtaagtggac gacccggtca taccttctta agttcgccct

         1801 tcctcccttt atttcagatt caatctgact tacctattct acccaagcat ccaaatgaaa

         1861 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc

         1921 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga

         1981 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat

         2041 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggag

         2101 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac

         2161 ctgcctgaaa ccgaactgcc cgctgttctc cagccggtcg cggaggccat ggatgcgatc

         2221 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt

         2281 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg

         2341 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg

         2401 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc atgcggattt cggctccaac

         2461 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc

         2521 ggggattccc aatacgaggt cgccaacatc ctcttctgga ggccgtggtt ggcttgtatg

         2581 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcgcctc

         2641 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat

         2701 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg

         2761 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta

         2821 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag

         2881 agtagatgcc gaccgggaac cagttaacgt cgacctcgag ggggggcccg gtaccagctt

         2941 ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat agctgtttcc

         3001 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg

         3061 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc

         3121 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg

         3181 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc

         3241 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac

         3301 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa

         3361 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca

         3421 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc

         3481 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata

         3541 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta

         3601 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca

         3661 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga

         3721 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg

         3781 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg

         3841 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg

         3901 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag

         3961 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa

         4021 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat

         4081 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc

         4141 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc

         4201 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc

         4261 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc

         4321 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc

         4381 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt

         4441 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc

         4501 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa

         4561 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt

         4621 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg

         4681 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc

         4741 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa

         4801 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt

         4861 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt

         4921 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag

         4981 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta

         5041 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat

         5101 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg

         5161 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc

         5221 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt

         5281 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc

         5341 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg

         5401 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga

         5461 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg

         5521 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg

         5581 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg

         5641 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt

         5701 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa

         5761 tacgactcac tatagggcga attggagctc

    //

     

    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.