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.