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p46815 empty yeast backbone.gb
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p46815 empty yeast backbone.gb
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LOCUS 6981 bp DNA circular GCC
DEFINITION addgene-plasmid-46815-sequence-66300
ACCESSION
SOURCE
ORGANISM
COMMENT This file was created using Genome Compiler.
FEATURES Location/Qualifiers
source 1..6981
/organism=""
terminator 2..167
/label="ADH1 terminator"
/locus_tag=""
/note="transcription terminator for the S. cerevisiae alcoh
ol dehydrogenase 1 (ADH1) gene"
/product=""
CDS complement(315..338)
/label="FLAG"
/locus_tag=""
/product="FLAG® epitope tag, followed by an enterokinase cl
eavage site"
/translation="DYKDDDDK"
promoter 1354..1760
/label="TEF1 promoter"
/locus_tag=""
/note="promoter for EF-1α"
/product=""
promoter 1780..1798
/label="T7 promoter"
/locus_tag=""
/note="promoter for bacteriophage T7 RNA polymerase"
/product=""
CDS 1816..1845
/label="Myc"
/locus_tag=""
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/translation="EQKLISEEDL"
terminator 1875..2064
/label="CYC1 terminator"
/locus_tag=""
/note="transcription terminator for CYC1"
/product=""
rep_origin complement(2307..2895)
/label="ori"
/locus_tag=""
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of rep
lication"
/product=""
CDS complement(3066..3926)
/label="AmpR"
/locus_tag=""
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/product="β-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
promoter complement(3927..4031)
/label="AmpR promoter"
/locus_tag=""
/product=""
rep_origin 4058..5400
/label="2μ ori"
/locus_tag=""
/note="yeast 2μ plasmid origin of replication"
/product=""
protein_bind complement(5420..5453)
/label="loxP"
/locus_tag=""
/note="Cre-mediated recombination occurs in the 8-bp core s
equence (GCATACAT)."
/product=""
promoter 5508..5851
/label="TEF promoter"
/locus_tag=""
/note="Ashbya gossypii TEF promoter"
/product=""
CDS 5852..6661
/label="KanR"
/locus_tag=""
/note="confers resistance to kanamycin"
/product="aminoglycoside phosphotransferase"
/translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGK
PDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVD
ASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRV
GIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
terminator 6667..6864
/label="TEF terminator"
/locus_tag=""
/note="Ashbya gossypii TEF terminator"
/product=""
protein_bind complement(6927..6960)
/label="loxP"
/locus_tag=""
/note="Cre-mediated recombination occurs in the 8-bp core s
equence (GCATACAT)."
/product=""
CDS complement(3066..3857)
/label="Segment of AmpR"
/locus_tag=""
/product=""
/translation="HPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMM
STFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAI
TMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVA
MATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSR
GIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
CDS complement(3858..3926)
/label="signal sequence"
/locus_tag=""
/product=""
/translation="MSIQHFRVALIPFFAAFCLPVFA"
protein_bind complement(5021..5068)
/label="FRT"
/locus_tag=""
/note="FLP-mediated recombination occurs in the 8-bp core s
equence TCTAGAAA (Turan and Bode, 2011)."
/product=""
CDS 5508..6864
/label="kanMX"
/locus_tag=""
/note="yeast selectable marker conferring kanamycin resista
nce (Wach et al., 1994)"
/product=""
/translation="DMEAQNTLLDSLDVRSSGACDCRPYI*PIHPHV*SFASIHFDGR
TARSKNYGSSLQTCEQGNAPLTDALNCPHAAPL*RNIKG*DLPLRFFFHILPFKILLG
YSSHITSEHKQPWVRKRLTFRGRD*IPTWMLIYMGINGLAIMSGNQVRQSIDCMGSPM
RQSCF*NMAKVALPMMLQMRWSD*TG*RNLCLFRPSSILSVLLMMHGYSPLRSPAKQH
SRY*KNILIQVKILLMRWQCSCAGCIRFLFVIVLLTAIAYFVSLRRNHE*ITVWLMRV
ILMTSVMAGLLNKSGKKCISFCHSHRIQSSLMVISHLITLFLTRGN**VVLMLDESES
QTDTRILPSYGTASVSFLLHYRNGFFKNMVLIILI*INCSFI*CSMSFSNQY*Q*KDS
CFQELVICIVFLYCSCSILIKC*RDLYFFSPRHHLPRCEVKCAESNIMRQSYVNAGRY
T"
ORIGIN
1 ggagcgacct catgctatac ctgagaaagc aacctgacct acaggaaaga gttactcaag
61 aataagaatt ttcgttttaa aacctaagag tcactttaaa atttgtatac acttattttt
121 tttataactt atttaataat aaaaatcata aatcataaga aattcgctta tttagaagtg
181 tcaacaacgt atctaccaac gatttgaccc ttttccatct tttcgtaaat ttctggcaag
241 gtagacaagc cgacaacctt gattggagac ttgaccaaac ctctggcgaa gaattgttaa
301 ttaagagctc agatcttatc gtcgtcatcc ttgtaatcca tcgatactag tgcggccgct
361 tgttttatat ttgttgtaaa aagtagataa ttacttcctt gatgatctgt aaaaaagaga
421 aaaagaaagc atctaagaac ttgaaaaact acgaattaga aaagaccaaa tatgtatttc
481 ttgcattgac caatttatgc aagtttatat atatgtaaat gtaagtttca cgaggttcta
541 ctaaactaaa ccaccccctt ggttagaaga aaagagtgtg tgagaacagg ctgttgttgt
601 cacacgattc ggacaattct gtttgaaaga gagagagtaa cagtacgatc gaacgaactt
661 tgctctggag atcacagtgg gcatcatagc atgtggtact aaaccctttc ccgccattcc
721 agaaccttcg attgcttgtt acaaaacctg tgagccgtcg ctaggacctt gttgtgtgac
781 gaaattggaa gctgcaatca ataggaagac aggaagtcga gcgtgtctgg gttttttcag
841 ttttgttctt tttgcaaaca aatcacgagc gacggtaatt tctttctcga taagaggcca
901 cgtgctttat gagggtaaca tcaattcaag aaggagggaa acacttcctt tttctggccc
961 tgataatagt atgagggtga agccaaaata aaggattcgc gcccaaatcg gcatctttaa
1021 atgcaggtat gcgatagttc ctcactcttt ccttactcac gagtaattct tgcaaatgcc
1081 tattatgcag atgttataat atctgtgcgt cttgagttga agtcaggaat ctaaaataaa
1141 aattaaggtt aataaaaaga ggaaagaaaa aaaaattaat cgatttacag aaacttgcac
1201 actaaaaata cacaactaaa agcaattaca gtatgggaag tcatcgacgt tatctctact
1261 atagtatatt atcatttcta ttattatcct gctcagtggt acttgcaaaa caagataaga
1321 ccccattctt tgaaggtact tccaggccgg ccgcacacac catagcttca aaatgtttct
1381 actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac cacttcaaaa
1441 cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc gttaattacc
1501 cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa
1561 aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt tgattttttt
1621 ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt
1681 tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa
1741 gcatagcaat ctaatctaag ttttaattac aaggatccgt aatacgactc actatagggc
1801 ccgggcgtcg acatggaaca gaagttgatt tccgaagaag acctcgagta agcttggtac
1861 cgcggctagc taagatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
1921 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
1981 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
2041 gagaaggttt tgggacgctc gaagatccag ctgcattaat gaatcggcca acgcgcgggg
2101 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
2161 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
2221 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
2281 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
2341 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
2401 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
2461 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
2521 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
2581 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
2641 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
2701 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
2761 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
2821 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
2881 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
2941 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
3001 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
3061 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
3121 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
3181 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
3241 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
3301 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
3361 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
3421 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
3481 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
3541 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
3601 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
3661 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
3721 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
3781 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
3841 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
3901 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
3961 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
4021 ggggttccgc gcacatttcc ccgaaaagtg ccacctgaac gaagcatctg tgcttcattt
4081 tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
4141 ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc
4201 atttttgtaa aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
4261 tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat
4321 acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct
4381 tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca
4441 ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa
4501 aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt
4561 caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa
4621 cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta
4681 ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac
4741 tctatgaata gttcttacta caattttttt gtctaaagag taatactaga gataaacata
4801 aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt
4861 atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga
4921 agcggtattc gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa
4981 agtgcgtctt cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag
5041 agaataggaa cttcggaata ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa
5101 tgcaacgcga gctgcgcaca tacagctcac tgttcacgtc gcacctatat ctgcgtgttg
5161 cctgtatata tatatacatg agaagaacgg catagtgcgt gtttatgctt aaatgcgtac
5221 ttatatgcgt ctatttatgt aggatgaaag gtagtctagt acctcctgtg atattatccc
5281 attccatgcg gggtatcgta tgcttccttc agcactaccc tttagctgtt ctatatgctg
5341 ccactcctca attggattag tctcatcctt caatgctatc atttcctttg atattggatc
5401 atggtagaca acccttaata taacttcgta taatgtatgc tatacgaagt tattaggtct
5461 agagatctgt ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc
5521 agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc
5581 cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc
5641 gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc
5701 ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt
5761 aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca
5821 gttctcacat cacatccgaa cataaacaac catgggtaag gaaaagactc acgtttcgag
5881 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
5941 tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt
6001 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
6061 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
6121 tgatgcatgg ttactcacca ctgcgatccc cggcaaaaca gcattccagg tattagaaga
6181 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
6241 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
6301 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
6361 ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc
6421 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
6481 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
6541 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
6601 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
6661 atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
6721 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
6781 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
6841 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtgtcg
6901 aaaacgagct ctcgagaacc cttaatataa cttcgtataa tgtatgctat acgaagttat
6961 taggtgatat cagatccact a
//