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.