Catalog No. PVT11277
Packing 2ug
P414-TEF1p-Cas9-CYC1t is a yeast yeast gene editing Cas9 expression plasmid.
LOCUS Exported 9524 bp ds-DNA circular SYN 06-JAN-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..9524
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 69..572
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 609..713
/gene="bla"
/label=AmpR promoter
CDS 714..1574
/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 1745..2333
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 2792..3190
/gene="S. cerevisiae TEF1"
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
CDS 3222..7325
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 7338..7358
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
terminator 7374..7621
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
rep_origin 7829..8284
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(8383..9057)
/codon_start=1
/gene="S. cerevisiae TRP1"
/product="phosphoribosylanthranilate isomerase, required
for tryptophan biosynthesis"
/label=TRP1
/note="yeast auxotrophic marker"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
promoter complement(9058..9338)
/gene="S. cerevisiae TRP1"
/label=TRP1 promoter
ORIGIN
1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa
121 tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta
181 aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa
241 tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag
301 atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt
361 ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa
421 aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta
481 ttttttcttt aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa
541 tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg
601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
1861 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
2521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
2581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tacctcactc
2641 attaggcacc ccaggcttta cactttatgc ttccggctcc tatgttgtgt ggaattgtga
2701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
2761 accctcacta aagggaacaa aagctggagc tcatagcttc aaaatgtttc tactcctttt
2821 ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc
2881 acagcatact aaatttcccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa
2941 ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa
3001 aaatttttat cacgtttctt tttcttgaaa attttttttt tgattttttt ctctttcgat
3061 gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt tcagtttcat
3121 ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa gcatagcaat
3181 ctaatctaag ttttctagaa ctagtggatc ccccgggaaa aatggacaag aagtactcca
3241 ttgggctcga tatcggcaca aacagcgtcg gttgggccgt cattacggac gagtacaagg
3301 tgccgagcaa aaaattcaaa gttctgggca ataccgatcg ccacagcata aagaagaacc
3361 tcattggcgc cctcctgttc gactccgggg agacggccga agccacgcgg ctcaaaagaa
3421 cagcacggcg cagatatacc cgcagaaaga atcggatctg ctacctgcag gagatcttta
3481 gtaatgagat ggctaaggtg gatgactctt tcttccatag gctggaggag tcctttttgg
3541 tggaggagga taaaaagcac gagcgccacc caatctttgg caatatcgtg gacgaggtgg
3601 cgtaccatga aaagtaccca accatatatc atctgaggaa gaagcttgta gacagtactg
3661 ataaggctga cttgcggttg atctatctcg cgctggcgca tatgatcaaa tttcggggac
3721 acttcctcat cgagggggac ctgaacccag acaacagcga tgtcgacaaa ctctttatcc
3781 aactggttca gacttacaat cagcttttcg aagagaaccc gatcaacgca tccggagttg
3841 acgccaaagc aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg
3901 cacagctccc tggggagaag aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg
3961 ggctgacccc caactttaaa tctaacttcg acctggccga agatgccaag cttcaactga
4021 gcaaagacac ctacgatgat gatctcgaca atctgctggc ccagatcggc gaccagtacg
4081 cagacctttt tttggcggca aagaacctgt cagacgccat tctgctgagt gatattctgc
4141 gagtgaacac ggagatcacc aaagctccgc tgagcgctag tatgatcaag cgctatgatg
4201 agcaccacca agacttgact ttgctgaagg cccttgtcag acagcaactg cctgagaagt
4261 acaaggaaat tttcttcgat cagtctaaaa atggctacgc cggatacatt gacggcggag
4321 caagccagga ggaattttac aaatttatta agcccatctt ggaaaaaatg gacggcaccg
4381 aggagctgct ggtaaagctt aacagagaag atctgttgcg caaacagcgc actttcgaca
4441 atggaagcat cccccaccag attcacctgg gcgaactgca cgctatcctc aggcggcaag
4501 aggatttcta cccctttttg aaagataaca gggaaaagat tgagaaaatc ctcacatttc
4561 ggatacccta ctatgtaggc cccctcgccc ggggaaattc cagattcgcg tggatgactc
4621 gcaaatcaga agagaccatc actccctgga acttcgagga agtcgtggat aagggggcct
4681 ctgcccagtc cttcatcgaa aggatgacta actttgataa aaatctgcct aacgaaaagg
4741 tgcttcctaa acactctctg ctgtacgagt acttcacagt ttataacgag ctcaccaagg
4801 tcaaatacgt cacagaaggg atgagaaagc cagcattcct gtctggagag cagaagaaag
4861 ctatcgtgga cctcctcttc aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag
4921 actatttcaa aaagattgaa tgtttcgact ctgttgaaat cagcggagtg gaggatcgct
4981 tcaacgcatc cctgggaacg tatcacgatc tcctgaaaat cattaaagac aaggacttcc
5041 tggacaatga ggagaacgag gacattcttg aggacattgt cctcaccctt acgttgtttg
5101 aagataggga gatgattgaa gaacgcttga aaacttacgc tcatctcttc gacgacaaag
5161 tcatgaaaca gctcaagagg cgccgatata caggatgggg gcggctgtca agaaaactga
5221 tcaatgggat ccgagacaag cagagtggaa agacaatcct ggattttctt aagtccgatg
5281 gatttgccaa ccggaacttc atgcagttga tccatgatga ctctctcacc tttaaggagg
5341 acatccagaa agcacaagtt tctggccagg gggacagtct tcacgagcac atcgctaatc
5401 ttgcaggtag cccagctatc aaaaagggaa tactgcagac cgttaaggtc gtggatgaac
5461 tcgtcaaagt aatgggaagg cataagcccg agaatatcgt tatcgagatg gcccgagaga
5521 accaaactac ccagaaggga cagaagaaca gtagggaaag gatgaagagg attgaagagg
5581 gtataaaaga actggggtcc caaatcctta aggaacaccc agttgaaaac acccagcttc
5641 agaatgagaa gctctacctg tactacctgc agaacggcag ggacatgtac gtggatcagg
5701 aactggacat caatcggctc tccgactacg acgtggatca tatcgtgccc cagtcttttc
5761 tcaaagatga ttctattgat aataaagtgt tgacaagatc cgataaaaat agagggaaga
5821 gtgataacgt cccctcagaa gaagttgtca agaaaatgaa aaattattgg cggcagctgc
5881 tgaacgccaa actgatcaca caacggaagt tcgataatct gactaaggct gaacgaggtg
5941 gcctgtctga gttggataaa gccggcttca tcaaaaggca gcttgttgag acacgccaga
6001 tcaccaagca cgtggcccaa attctcgatt cacgcatgaa caccaagtac gatgaaaatg
6061 acaaactgat tcgagaggtg aaagttatta ctctgaagtc taagctggtc tcagatttca
6121 gaaaggactt tcagttttat aaggtgagag agatcaacaa ttaccaccat gcgcatgatg
6181 cctacctgaa tgcagtggta ggcactgcac ttatcaaaaa atatcccaag cttgaatctg
6241 aatttgttta cggagactat aaagtgtacg atgttaggaa aatgatcgca aagtctgagc
6301 aggaaatagg caaggccacc gctaagtact tcttttacag caatattatg aattttttca
6361 agaccgagat tacactggcc aatggagaga ttcggaagcg accacttatc gaaacaaacg
6421 gagaaacagg agaaatcgtg tgggacaagg gtagggattt cgcgacagtc cggaaggtcc
6481 tgtccatgcc gcaggtgaac atcgttaaaa agaccgaagt acagaccgga ggcttctcca
6541 aggaaagtat cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa aaagattggg
6601 accccaagaa atacggcgga ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg
6661 ccaaagtgga gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca
6721 caatcatgga gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat
6781 ataaagaggt caaaaaagac ctcatcatta agcttcccaa gtactctctc tttgagcttg
6841 aaaacggccg gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg
6901 cactgccctc taaatacgtt aatttcttgt atctggccag ccactatgaa aagctcaaag
6961 ggtctcccga agataatgag cagaagcagc tgttcgtgga acaacacaaa cactaccttg
7021 atgagatcat cgagcaaata agcgaattct ccaaaagagt gatcctcgcc gacgctaacc
7081 tcgataaggt gctttctgct tacaataagc acagggataa gcccatcagg gagcaggcag
7141 aaaacattat ccacttgttt actctgacca acttgggcgc gcctgcagcc ttcaagtact
7201 tcgacaccac catagacaga aagcggtaca cctctacaaa ggaggtcctg gacgccacac
7261 tgattcatca gtcaattacg gggctctatg aaacaagaat cgacctctct cagctcggtg
7321 gagacagcag ggctgacccc aagaagaaga ggaaggtgtg atctcttctc gagtcatgta
7381 attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga
7441 aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt
7501 aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg
7561 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg
7621 cggccggtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
7681 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
7741 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
7801 ttgcgcagcc tgaatggcga atggcgcgac gcgccctgta gcggcgcatt aagcgcggcg
7861 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
7921 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
7981 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
8041 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
8101 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
8161 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
8221 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
8281 atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataggca
8341 agtgcacaaa caatacttaa ataaatacta ctcagtaata acctatttct tagcattttt
8401 gacgaaattt gctattttgt tagagtcttt tacaccattt gtctccacac ctccgcttac
8461 atcaacacca ataacgccat ttaatctaag cgcatcacca acattttctg gcgtcagtcc
8521 accagctaac ataaaatgta agctttcggg gctctcttgc cttccaaccc agtcagaaat
8581 cgagttccaa tccaaaagtt cacctgtccc acctgcttct gaatcaaaca agggaataaa
8641 cgaatgaggt ttctgtgaag ctgcactgag tagtatgttg cagtcttttg gaaatacgag
8701 tcttttaata actggcaaac cgaggaactc ttggtattct tgccacgact catctccatg
8761 cagttggacg atatcaatgc cgtaatcatt gaccagagcc aaaacatcct ccttaggttg
8821 attacgaaac acgccaacca agtatttcgg agtgcctgaa ctatttttat atgcttttac
8881 aagacttgaa attttccttg caataaccgg gtcaattgtt ctctttctat tgggcacaca
8941 tataataccc agcaagtcag catcggaatc tagagcacat tctgcggcct ctgtgctctg
9001 caagccgcaa actttcacca atggaccaga actacctgtg aaattaataa cagacatact
9061 ccaagctgcc tttgtgtgct taatcacgta tactcacgtg ctcaatagtc accaatgccc
9121 tccctcttgg ccctctcctt ttcttttttc gaccgaatta attcttaatc ggcaaaaaaa
9181 gaaaagctcc ggatcaagat tgtacgtaag gtgacaagct atttttcaat aaagaatatc
9241 ttccactact gccatctggc gtcataactg caaagtacac atatattacg atgctgtcta
9301 ttaaatgctt cctatattat atatatagta atgtcgttta tggtgcactc tcagtacaat
9361 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
9421 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
9481 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcga
//
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