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| clotech |
| pGADT7 |
| 酿酒酵母表达载体 |
| -- |
| ADH1 |
| 多克隆位点，限制性内切酶 |
| 7988bp |
| T7: TAATACGACTCACTATAGGG |
| -- |
| -- |
| Ampicillin |
| -- |
| -- |
| 630442 |
| -- |
| -- |
| 非病毒 |
载体质粒图谱和多克隆位点信息
载体简介
pGADT7 AD is a yeast expression vector that is designed to express a protein of interest
fused to a GAL4 activation domain (AD; amino acids 768–881). Transcription of the GAL4 AD
fusion is driven by the constitutively active ADH1 promoter (PADH1), and is terminated at the
ADH1 transcription termination signal (TADH1). The GAL4 AD fusion contains an N-terminal
SV40 nuclear localization signal (SV40 NLS; 1) that targets the protein to the yeast nucleus,
and a hemagglutinin epitope tag (HA Tag), located between the GAL4 AD and the protein
of interest, that allows the protein to be easily detected with HA-tag antibodies.
The T7 promoter (PT7), located just upstream of the HA tag sequence, allows in vitro transcription
and translation of the HA-tagged protein of interest (without the GAL4 AD and
the SV40 NLS). pGADT7 AD replicates autonomously in both E. coli and S. cerevisiae from
the pUC and 2 ori, respectively. The vector also contains an ampicillin resistance gene
(Ampr) for selection in E. coli and a LEU2 nutritional marker for selection in yeast.
fused to a GAL4 activation domain (AD; amino acids 768–881). Transcription of the GAL4 AD
fusion is driven by the constitutively active ADH1 promoter (PADH1), and is terminated at the
ADH1 transcription termination signal (TADH1). The GAL4 AD fusion contains an N-terminal
SV40 nuclear localization signal (SV40 NLS; 1) that targets the protein to the yeast nucleus,
and a hemagglutinin epitope tag (HA Tag), located between the GAL4 AD and the protein
of interest, that allows the protein to be easily detected with HA-tag antibodies.
The T7 promoter (PT7), located just upstream of the HA tag sequence, allows in vitro transcription
and translation of the HA-tagged protein of interest (without the GAL4 AD and
the SV40 NLS). pGADT7 AD replicates autonomously in both E. coli and S. cerevisiae from
the pUC and 2 ori, respectively. The vector also contains an ampicillin resistance gene
(Ampr) for selection in E. coli and a LEU2 nutritional marker for selection in yeast.
载体序列
LOCUS pGAD T7 7988 bp DNA SYN 23-Nov-2013
DEFINITION pGAD T7
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..7988
/organism="pGAD T7"
/mol_type="other DNA"
promoter 1079..1476
/label="yADH1_promoter"
misc_feature 1501..1557
/label="SV40_NLS"
misc_feature 1492..1899
/label="Gal4_AD"
misc_feature 1561..1899
/label="Gal4_AD"
misc_feature 1825..1842
/label="Gal4_AD_primer"
promoter 1905..1923
/label="T7_promoter"
misc_feature complement(2295..2318)
/label="pJG4_5_rev_primer"
terminator 2279..2605
/label="yADH2_terminator"
misc_feature complement(2471..2495)
/label="pHybLex_rev_primer"
terminator 2416..2595
/label="yADH1_terminator"
CDS complement(2720..3814)
/label="LEU2(variant)"
/gene="LEU2(variant)"
/note="ORF frame 2"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENH
LIGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQL
YANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSE
QYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLK
VQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPD
KNTAFGLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLD
AGIRTGDLGGSNSTTEVGDAVAEEVKKILA*"
gene complement(2720..3814)
/label="LEU2(variant)"
/gene="LEU2(variant)"
misc_feature complement(4258..4276)
/label="M13_reverse_primer"
misc_feature complement(4275..4297)
/label="M13_pUC_rev_primer"
promoter complement(4311..4340)
/label="lac_promoter"
misc_feature complement(4431..4464)
/label="loxP"
misc_feature complement(4542..4575)
/label="loxP"
rep_origin complement(4800..5419)
/label="pBR322_origin"
CDS complement(5574..6434)
/label="Ampicillin"
/gene="Ampicillin"
/note="ORF frame 3"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
gene complement(5574..6434)
/label="Ampicillin"
/gene="Ampicillin"
promoter complement(6476..6504)
/label="AmpR_promoter"
misc_feature complement(6663..6685)
/label="pGEX_3_primer"
rep_origin 6821..7985
/label="2micron2_origin"
ORIGIN
1 TGCATGCCTG CAGGTCGAGA TCCGGGATCG AAGAAATGAT GGTAAATGAA ATAGGAAATC
61 AAGGAGCATG AAGGCAAAAG ACAAATATAA GGGTCGAACG AAAAATAAAG TGAAAAGTGT
121 TGATATGATG TATTTGGCTT TGCGGCGCCG AAAAAACGAG TTTACGCAAT TGCACAATCA
181 TGCTGACTCT GTGGCGGACC CGCGCTCTTG CCGGCCCGGC GATAACGCTG GGCGTGAGGC
241 TGTGCCCGGC GGAGTTTTTT GCGCCTGCAT TTTCCAAGGT TTACCCTGCG CTAAGGGGCG
301 AGATTGGAGA AGCAATAAGA ATGCCGGTTG GGGTTGCGAT GATGACGACC ACGACAACTG
361 GTGTCATTAT TTAAGTTGCC GAAAGAACCT GAGTGCATTT GCAACATGAG TATACTAGAA
421 GAATGAGCCA AGACTTGCGA GACGCGAGTT TGCCGGTGGT GCGAACAATA GAGCGACCAT
481 GACCTTGAAG GTGAGACGCG CATAACCGCT AGAGTACTTT GAAGAGGAAA CAGCAATAGG
541 GTTGCTACCA GTATAAATAG ACAGGTACAT ACAACACTGG AAATGGTTGT CTGTTTGAGT
601 ACGCTTTCAA TTCATTTGGG TGTGCACTTT ATTATGTTAC AATATGGAAG GGAACTTTAC
661 ACTTCTCCTA TGCACATATA TTAATTAAAG TCCAATGCTA GTAGAGAAGG GGGGTAACAC
721 CCCTCCGCGC TCTTTTCCGA TTTTTTTCTA AACCGTGGAA TATTTCGGAT ATCCTTTTGT
781 TGTTTCCGGG TGTACAATAT GGACTTCCTC TTTTCTGGCA ACCAAACCCA TACATCGGGA
841 TTCCTATAAT ACCTTCGTTG GTCTCCCTAA CATGTAGGTG GCGGAGGGGA GATATACAAT
901 AGAACAGATA CCAGACAAGA CATAATGGGC TAAACAAGAC TACACCAATT ACACTGCCTC
961 ATTGATGGTG GTACATAACG AACTAATACT GTAGCCCTAG ACTTGATAGC CATCATCATA
1021 TCGAAGTTTC ACTACCCTTT TTCCATTTGC CATCTATTGA AGTAATAATA GGCGCATGCA
1081 ACTTCTTTTC TTTTTTTTTC TTTTCTCTCT CCCCCGTTGT TGTCTCACCA TATCCGCAAT
1141 GACAAAAAAA ATGATGGAAG ACACTAAAGG AAAAAATTAA CGACAAAGAC AGCACCAACA
1201 GATGTCGTTG TTCCAGAGCT GATGAGGGGT ATCTCGAAGC ACACGAAACT TTTTCCTTCC
1261 TTCATTCACG CACACTACTC TCTAATGAGC AACGGTATAC GGCCTTCCTT CCAGTTACTT
1321 GAATTTGAAA TAAAAAAAAG TTTGCTGTCT TGCTATCAAG TATAAATAGA CCTGCAATTA
1381 TTAATCTTTT GTTTCCTCGT CATTGTTCTC GTTCCCTTTC TTCCTTGTTT CTTTTTCTGC
1441 ACAATATTTC AAGCTATACC AAGCATACAA TCAACTCCAA GCTTTGCAAA GATGGATAAA
1501 GCGGAATTAA TTCCCGAGCC TCCAAAAAAG AAGAGAAAGG TCGAATTGGG TACCGCCGCC
1561 AATTTTAATC AAAGTGGGAA TATTGCTGAT AGCTCATTGT CCTTCACTTT CACTAACAGT
1621 AGCAACGGTC CGAACCTCAT AACAACTCAA ACAAATTCTC AAGCGCTTTC ACAACCAATT
1681 GCCTCCTCTA ACGTTCATGA TAACTTCATG AATAATGAAA TCACGGCTAG TAAAATTGAT
1741 GATGGTAATA ATTCAAAACC ACTGTCACCT GGTTGGACGG ACCAAACTGC GTATAACGCG
1801 TTTGGAATCA CTACAGGGAT GTTTAATACC ACTACAATGG ATGATGTATA TAACTATCTA
1861 TTCGATGATG AAGATACCCC ACCAAACCCA AAAAAAGAGA TCTTTAATAC GACTCACTAT
1921 AGGGCGAGCG CCGCCATGGA GTACCCATAC GACGTACCAG ATTACGCTCA TATGGCCATG
1981 GAGGCCAGTG AATTCCACCC GGGTGGGCAT CGATACGGGA TCCATCGAGC TCGAGCTGCA
2041 GATGAATCGT AGATACTGAA AAACCCCGCA AGTTCACTTC AACTGTGCAT CGTGCACCAT
2101 CTCAATTTCT TTCATTTATA CATCGTTTTG CCTTCTTTTA TGTAACTATA CTCCTCTAAG
2161 TTTCAATCTT GGCCATGTAA CCTCTGATCT ATAGAATTTT TTAAATGACT AGAATTAATG
2221 CCCATCTTTT TTTTGGACCT AAATTCTTCA TGAAAATATA TTACGAGGGC TTATTCAGAA
2281 GCTTTGGACT TCTTCGCCAG AGGTTTGGTC AAGTCTCCAA TCAAGGTTGT CGGCTTGTCT
2341 ACCTTGCCAG AAATTTACGA AAAGATGGAA AAGGGTCAAA TCGTTGGTAG ATACGTTGTT
2401 GACACTTCTA AATAAGCGAA TTTCTTATGA TTTATGATTT TTATTATTAA ATAAGTTATA
2461 AAAAAAATAA GTGTATACAA ATTTTAAAGT GACTCTTAGG TTTTAAAACG AAAATTCTTA
2521 TTCTTGAGTA ACTCTTTCCT GTAGGTCAGG TTGCTTTCTC AGGTATAGCA TGAGGTCGCT
2581 CTTATTGACC ACACCTCTAC CGGCCGGTCG AAATTCCCCT ACCCTATGAA CATATTCCAT
2641 TTTGTAATTT CGTGTCGTTT CTATTATGAA TTTCATTTAT AAAGTTTATG TACAAATATC
2701 ATAAAAAAAG AGAATCTTTT TAAGCAAGGA TTTTCTTAAC TTCTTCGGCG ACAGCATCAC
2761 CGACTTCGGT GGTACTGTTG GAACCACCTA AATCACCAGT TCTGATACCT GCATCCAAAA
2821 CCTTTTTAAC TGCATCTTCA ATGGCCTTAC CTTCTTCAGG CAAGTTCAAT GACAATTTCA
2881 ACATCATTGC AGCAGACAAG ATAGTGGCGA TAGGGTTGAC CTTATTCTTT GGCAAATCTG
2941 GAGCAGAACC GTGGCATGGT TCGTACAAAC CAAATGCGGT GTTCTTGTCT GGCAAAGAGG
3001 CCAAGGACGC AGATGGCAAC AAACCCAAGG AACCTGGGAT AACGGAGGCT TCATCGGAGA
3061 TGATATCACC AAACATGTTG CTGGTGATTA TAATACCATT TAGGTGGGTT GGGTTCTTAA
3121 CTAGGATCAT GGCGGCAGAA TCAATCAATT GATGTTGAAC CTTCAATGTA GGAAATTCGT
3181 TCTTGATGGT TTCCTCCACA GTTTTTCTCC ATAATCTTGA AGAGGCCAAA ACATTAGCTT
3241 TATCCAAGGA CCAAATAGGC AATGGTGGCT CATGTTGTAG GGCCATGAAA GCGGCCATTC
3301 TTGTGATTCT TTGCACTTCT GGAACGGTGT ATTGTTCACT ATCCCAAGCG ACACCATCAC
3361 CATCGTCTTC CTTTCTCTTA CCAAAGTAAA TACCTCCCAC TAATTCTCTG ACAACAACGA
3421 AGTCAGTACC TTTAGCAAAT TGTGGCTTGA TTGGAGATAA GTCTAAAAGA GAGTCGGATG
3481 CAAAGTTACA TGGTCTTAAG TTGGCGTACA ATTGAAGTTC TTTACGGATT TTTAGTAAAC
3541 CTTGTTCAGG TCTAACACTA CCTGTACCCC ATTTAGGACC ACCCACAGCA CCTAACAAAA
3601 CGGCATCAAC CTTCTTGGAG GCTTCCAGCG CCTCATCTGG AAGTGGGACA CCTGTAGCGT
3661 CGATAGCAGC ACCACCAATT AAATGATTTT CGAAATCGAA CTTGACATTG GAACGAACAT
3721 CAGAAATAGC TTTAAGAACC TTAATGGCTT CGGCTGTGAT TTCTTGACCA ACGTGGTCAC
3781 CTGGCAAAAC GACGATCTTC TTAGGGGCAG ACATTAGAAT GGTATATCCT TGAAATATAT
3841 ATATATATTG CTGAAATGTA AAAGGTAAGA AAAGTTAGAA AGTAAGACGA TTGCTAACCA
3901 CCTATTGGAA AAAACAATAG GTCCTTAAAT AATATTGTCA ACTTCAAGTA TTGTGATGCA
3961 AGCATTTAGT CATGAACGCT TCTCTATTCT ATATGAAAAG CCGGTTCCGG CGCTCTCACC
4021 TTTCCTTTTT CTCCCAATTT TTCAGTTGAA AAAGGTATAT GCGTCAGGCG ACCTCTGAAA
4081 TTAACAAAAA ATTTCCAGTC ATCGAATTTG ATTCTGTGCG ATAGCGCCCC TGTGTGTTCT
4141 CGTTATGTTG AGGAAAAAAA TAATGGTTGC TAAGAGATTC GAACTCTTGC ATCTTACGAT
4201 ACCTGAGTAT TCCCACAGTT GGGGATCTCG ACTCTAGCTA GAGGATCAAT TCGTAATCAT
4261 GGTCATAGCT GTTTCCTGTG TGAAATTGTT ATCCGCTCAC AATTCCACAC AACATACGAG
4321 CCGGAAGCAT AAAGTGTAAA GCCTGGGGTG CCTAATGAGT GAGCTAACTC ACATTAATTG
4381 CGTTGCGCTC ACTGCCCGCT TTCCAGTCGG GAAACCTGTC GTGCCAGCTG ATAACTTCGT
4441 ATAATGTATG CTATACGAAG TTATTAGGTC TGAAGAGGAG TTTACGTCCA GCCAAGCTAG
4501 CTTGGCTGCA GGTCGAGCGG CCGCGATCCG GAACCCTTAA TATAACTTCG TATAATGTAT
4561 GCTATACGAA GTTATCAGCT GCATTAATGA ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG
4621 CGTATTGGGC GCTCTTCCGC TTCCTCGCTC ACTGACTCGC TGCGCTCGGT CGTTCGGCTG
4681 CGGCGAGCGG TATCAGCTCA CTCAAAGGCG GTAATACGGT TATCCACAGA ATCAGGGGAT
4741 AACGCAGGAA AGAACATGTG AGCAAAAGGC CAGCAAAAGG CCAGGAACCG TAAAAAGGCC
4801 GCGTTGCTGG CGTTTTTCCA TAGGCTCCGC CCCCCTGACG AGCATCACAA AAATCGACGC
4861 TCAAGTCAGA GGTGGCGAAA CCCGACAGGA CTATAAAGAT ACCAGGCGTT TCCCCCTGGA
4921 AGCTCCCTCG TGCGCTCTCC TGTTCCGACC CTGCCGCTTA CCGGATACCT GTCCGCCTTT
4981 CTCCCTTCGG GAAGCGTGGC GCTTTCTCAT AGCTCACGCT GTAGGTATCT CAGTTCGGTG
5041 TAGGTCGTTC GCTCCAAGCT GGGCTGTGTG CACGAACCCC CCGTTCAGCC CGACCGCTGC
5101 GCCTTATCCG GTAACTATCG TCTTGAGTCC AACCCGGTAA GACACGACTT ATCGCCACTG
5161 GCAGCAGCCA CTGGTAACAG GATTAGCAGA GCGAGGTATG TAGGCGGTGC TACAGAGTTC
5221 TTGAAGTGGT GGCCTAACTA CGGCTACACT AGAAGGACAG TATTTGGTAT CTGCGCTCTG
5281 CTGAAGCCAG TTACCTTCGG AAAAAGAGTT GGTAGCTCTT GATCCGGCAA ACAAACCACC
5341 GCTGGTAGCG GTGGTTTTTT TGTTTGCAAG CAGCAGATTA CGCGCAGAAA AAAAGGATCT
5401 CAAGAAGATC CTTTGATCTT TTCTACGGGG TCTGACGCTC AGTGGAACGA AAACTCACGT
5461 TAAGGGATTT TGGTCATGAG ATTATCAAAA AGGATCTTCA CCTAGATCCT TTTAAATTAA
5521 AAATGAAGTT TTAAATCAAT CTAAAGTATA TATGAGTAAA CTTGGTCTGA CAGTTACCAA
5581 TGCTTAATCA GTGAGGCACC TATCTCAGCG ATCTGTCTAT TTCGTTCATC CATAGTTGCC
5641 TGACTCCCCG TCGTGTAGAT AACTACGATA CGGGAGGGCT TACCATCTGG CCCCAGTGCT
5701 GCAATGATAC CGCGAGACCC ACGCTCACCG GCTCCAGATT TATCAGCAAT AAACCAGCCA
5761 GCCGGAAGGG CCGAGCGCAG AAGTGGTCCT GCAACTTTAT CCGCCTCCAT CCAGTCTATT
5821 AATTGTTGCC GGGAAGCTAG AGTAAGTAGT TCGCCAGTTA ATAGTTTGCG CAACGTTGTT
5881 GCCATTGCTA CAGGCATCGT GGTGTCACGC TCGTCGTTTG GTATGGCTTC ATTCAGCTCC
5941 GGTTCCCAAC GATCAAGGCG AGTTACATGA TCCCCCATGT TGTGCAAAAA AGCGGTTAGC
6001 TCCTTCGGTC CTCCGATCGT TGTCAGAAGT AAGTTGGCCG CAGTGTTATC ACTCATGGTT
6061 ATGGCAGCAC TGCATAATTC TCTTACTGTC ATGCCATCCG TAAGATGCTT TTCTGTGACT
6121 GGTGAGTACT CAACCAAGTC ATTCTGAGAA TAGTGTATGC GGCGACCGAG TTGCTCTTGC
6181 CCGGCGTCAA TACGGGATAA TACCGCGCCA CATAGCAGAA CTTTAAAAGT GCTCATCATT
6241 GGAAAACGTT CTTCGGGGCG AAAACTCTCA AGGATCTTAC CGCTGTTGAG ATCCAGTTCG
6301 ATGTAACCCA CTCGTGCACC CAACTGATCT TCAGCATCTT TTACTTTCAC CAGCGTTTCT
6361 GGGTGAGCAA AAACAGGAAG GCAAAATGCC GCAAAAAAGG GAATAAGGGC GACACGGAAA
6421 TGTTGAATAC TCATACTCTT CCTTTTTCAA TATTATTGAA GCATTTATCA GGGTTATTGT
6481 CTCATGAGCG GATACATATT TGAATGTATT TAGAAAAATA AACAAATAGG GGTTCCGCGC
6541 ACATTTCCCC GAAAAGTGCC ACCTGACGTC TAAGAAACCA TTATTATCAT GACATTAACC
6601 TATAAAAATA GGCGTATCAC GAGGCCCTTT CGTCTCGCGC GTTTCGGTGA TGACGGTGAA
6661 AACCTCTGAC ACATGCAGCT CCCGGAGACG GTCACAGCTT GTCTGTAAGC GGATGCCGGG
6721 AGCAGACAAG CCCGTCAGGG CGCGTCAGCG GGTGTTGGCG GGTGTCGGGG CTGGCTTAAC
6781 TATGCGGCAT CAGAGCAGAT TGTACTGAGA GTGCACCATA ACGCATTTAA GCATAAACAC
6841 GCACTATGCC GTTCTTCTCA TGTATATATA TATACAGGCA ACACGCAGAT ATAGGTGCGA
6901 CGTGAACAGT GAGCTGTATG TGCGCAGCTC GCGTTGCATT TTCGGAAGCG CTCGTTTTCG
6961 GAAACGCTTT GAAGTTCCTA TTCCGAAGTT CCTATTCTCT AGCTAGAAAG TATAGGAACT
7021 TCAGAGCGCT TTTGAAAACC AAAAGCGCTC TGAAGACGCA CTTTCAAAAA ACCAAAAACG
7081 CACCGGACTG TAACGAGCTA CTAAAATATT GCGAATACCG CTTCCACAAA CATTGCTCAA
7141 AAGTATCTCT TTGCTATATA TCTCTGTGCT ATATCCCTAT ATAACCTACC CATCCACCTT
7201 TCGCTCCTTG AACTTGCATC TAAACTCGAC CTCTACATTT TTTATGTTTA TCTCTAGTAT
7261 TACTCTTTAG ACAAAAAAAT TGTAGTAAGA ACTATTCATA GAGTGAATCG AAAACAATAC
7321 GAAAATGTAA ACATTTCCTA TACGTAGTAT ATAGAGACAA AATAGAAGAA ACCGTTCATA
7381 ATTTTCTGAC CAATGAAGAA TCATCAACGC TATCACTTTC TGTTCACAAA GTATGCGCAA
7441 TCCACATCGG TATAGAATAT AATCGGGGAT GCCTTTATCT TGAAAAAATG CACCCGCAGC
7501 TTCGCTAGTA ATCAGTAAAC GCGGGAAGTG GAGTCAGGCT TTTTTTATGG AAGAGAAAAT
7561 AGACACCAAA GTAGCCTTCT TCTAACCTTA ACGGACCTAC AGTGCAAAAA GTTATCAAGA
7621 GACTGCATTA TAGAGCGCAC AAAGGAGAAA AAAAGTAATC TAAGATGCTT TGTTAGAAAA
7681 ATAGCGCTCT CGGGATGCAT TTTTGTAGAA CAAAAAAGAA GTATAGATTC TTTGTTGGTA
DEFINITION pGAD T7
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..7988
/organism="pGAD T7"
/mol_type="other DNA"
promoter 1079..1476
/label="yADH1_promoter"
misc_feature 1501..1557
/label="SV40_NLS"
misc_feature 1492..1899
/label="Gal4_AD"
misc_feature 1561..1899
/label="Gal4_AD"
misc_feature 1825..1842
/label="Gal4_AD_primer"
promoter 1905..1923
/label="T7_promoter"
misc_feature complement(2295..2318)
/label="pJG4_5_rev_primer"
terminator 2279..2605
/label="yADH2_terminator"
misc_feature complement(2471..2495)
/label="pHybLex_rev_primer"
terminator 2416..2595
/label="yADH1_terminator"
CDS complement(2720..3814)
/label="LEU2(variant)"
/gene="LEU2(variant)"
/note="ORF frame 2"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENH
LIGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQL
YANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSE
QYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLK
VQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPD
KNTAFGLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLD
AGIRTGDLGGSNSTTEVGDAVAEEVKKILA*"
gene complement(2720..3814)
/label="LEU2(variant)"
/gene="LEU2(variant)"
misc_feature complement(4258..4276)
/label="M13_reverse_primer"
misc_feature complement(4275..4297)
/label="M13_pUC_rev_primer"
promoter complement(4311..4340)
/label="lac_promoter"
misc_feature complement(4431..4464)
/label="loxP"
misc_feature complement(4542..4575)
/label="loxP"
rep_origin complement(4800..5419)
/label="pBR322_origin"
CDS complement(5574..6434)
/label="Ampicillin"
/gene="Ampicillin"
/note="ORF frame 3"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
gene complement(5574..6434)
/label="Ampicillin"
/gene="Ampicillin"
promoter complement(6476..6504)
/label="AmpR_promoter"
misc_feature complement(6663..6685)
/label="pGEX_3_primer"
rep_origin 6821..7985
/label="2micron2_origin"
ORIGIN
1 TGCATGCCTG CAGGTCGAGA TCCGGGATCG AAGAAATGAT GGTAAATGAA ATAGGAAATC
61 AAGGAGCATG AAGGCAAAAG ACAAATATAA GGGTCGAACG AAAAATAAAG TGAAAAGTGT
121 TGATATGATG TATTTGGCTT TGCGGCGCCG AAAAAACGAG TTTACGCAAT TGCACAATCA
181 TGCTGACTCT GTGGCGGACC CGCGCTCTTG CCGGCCCGGC GATAACGCTG GGCGTGAGGC
241 TGTGCCCGGC GGAGTTTTTT GCGCCTGCAT TTTCCAAGGT TTACCCTGCG CTAAGGGGCG
301 AGATTGGAGA AGCAATAAGA ATGCCGGTTG GGGTTGCGAT GATGACGACC ACGACAACTG
361 GTGTCATTAT TTAAGTTGCC GAAAGAACCT GAGTGCATTT GCAACATGAG TATACTAGAA
421 GAATGAGCCA AGACTTGCGA GACGCGAGTT TGCCGGTGGT GCGAACAATA GAGCGACCAT
481 GACCTTGAAG GTGAGACGCG CATAACCGCT AGAGTACTTT GAAGAGGAAA CAGCAATAGG
541 GTTGCTACCA GTATAAATAG ACAGGTACAT ACAACACTGG AAATGGTTGT CTGTTTGAGT
601 ACGCTTTCAA TTCATTTGGG TGTGCACTTT ATTATGTTAC AATATGGAAG GGAACTTTAC
661 ACTTCTCCTA TGCACATATA TTAATTAAAG TCCAATGCTA GTAGAGAAGG GGGGTAACAC
721 CCCTCCGCGC TCTTTTCCGA TTTTTTTCTA AACCGTGGAA TATTTCGGAT ATCCTTTTGT
781 TGTTTCCGGG TGTACAATAT GGACTTCCTC TTTTCTGGCA ACCAAACCCA TACATCGGGA
841 TTCCTATAAT ACCTTCGTTG GTCTCCCTAA CATGTAGGTG GCGGAGGGGA GATATACAAT
901 AGAACAGATA CCAGACAAGA CATAATGGGC TAAACAAGAC TACACCAATT ACACTGCCTC
961 ATTGATGGTG GTACATAACG AACTAATACT GTAGCCCTAG ACTTGATAGC CATCATCATA
1021 TCGAAGTTTC ACTACCCTTT TTCCATTTGC CATCTATTGA AGTAATAATA GGCGCATGCA
1081 ACTTCTTTTC TTTTTTTTTC TTTTCTCTCT CCCCCGTTGT TGTCTCACCA TATCCGCAAT
1141 GACAAAAAAA ATGATGGAAG ACACTAAAGG AAAAAATTAA CGACAAAGAC AGCACCAACA
1201 GATGTCGTTG TTCCAGAGCT GATGAGGGGT ATCTCGAAGC ACACGAAACT TTTTCCTTCC
1261 TTCATTCACG CACACTACTC TCTAATGAGC AACGGTATAC GGCCTTCCTT CCAGTTACTT
1321 GAATTTGAAA TAAAAAAAAG TTTGCTGTCT TGCTATCAAG TATAAATAGA CCTGCAATTA
1381 TTAATCTTTT GTTTCCTCGT CATTGTTCTC GTTCCCTTTC TTCCTTGTTT CTTTTTCTGC
1441 ACAATATTTC AAGCTATACC AAGCATACAA TCAACTCCAA GCTTTGCAAA GATGGATAAA
1501 GCGGAATTAA TTCCCGAGCC TCCAAAAAAG AAGAGAAAGG TCGAATTGGG TACCGCCGCC
1561 AATTTTAATC AAAGTGGGAA TATTGCTGAT AGCTCATTGT CCTTCACTTT CACTAACAGT
1621 AGCAACGGTC CGAACCTCAT AACAACTCAA ACAAATTCTC AAGCGCTTTC ACAACCAATT
1681 GCCTCCTCTA ACGTTCATGA TAACTTCATG AATAATGAAA TCACGGCTAG TAAAATTGAT
1741 GATGGTAATA ATTCAAAACC ACTGTCACCT GGTTGGACGG ACCAAACTGC GTATAACGCG
1801 TTTGGAATCA CTACAGGGAT GTTTAATACC ACTACAATGG ATGATGTATA TAACTATCTA
1861 TTCGATGATG AAGATACCCC ACCAAACCCA AAAAAAGAGA TCTTTAATAC GACTCACTAT
1921 AGGGCGAGCG CCGCCATGGA GTACCCATAC GACGTACCAG ATTACGCTCA TATGGCCATG
1981 GAGGCCAGTG AATTCCACCC GGGTGGGCAT CGATACGGGA TCCATCGAGC TCGAGCTGCA
2041 GATGAATCGT AGATACTGAA AAACCCCGCA AGTTCACTTC AACTGTGCAT CGTGCACCAT
2101 CTCAATTTCT TTCATTTATA CATCGTTTTG CCTTCTTTTA TGTAACTATA CTCCTCTAAG
2161 TTTCAATCTT GGCCATGTAA CCTCTGATCT ATAGAATTTT TTAAATGACT AGAATTAATG
2221 CCCATCTTTT TTTTGGACCT AAATTCTTCA TGAAAATATA TTACGAGGGC TTATTCAGAA
2281 GCTTTGGACT TCTTCGCCAG AGGTTTGGTC AAGTCTCCAA TCAAGGTTGT CGGCTTGTCT
2341 ACCTTGCCAG AAATTTACGA AAAGATGGAA AAGGGTCAAA TCGTTGGTAG ATACGTTGTT
2401 GACACTTCTA AATAAGCGAA TTTCTTATGA TTTATGATTT TTATTATTAA ATAAGTTATA
2461 AAAAAAATAA GTGTATACAA ATTTTAAAGT GACTCTTAGG TTTTAAAACG AAAATTCTTA
2521 TTCTTGAGTA ACTCTTTCCT GTAGGTCAGG TTGCTTTCTC AGGTATAGCA TGAGGTCGCT
2581 CTTATTGACC ACACCTCTAC CGGCCGGTCG AAATTCCCCT ACCCTATGAA CATATTCCAT
2641 TTTGTAATTT CGTGTCGTTT CTATTATGAA TTTCATTTAT AAAGTTTATG TACAAATATC
2701 ATAAAAAAAG AGAATCTTTT TAAGCAAGGA TTTTCTTAAC TTCTTCGGCG ACAGCATCAC
2761 CGACTTCGGT GGTACTGTTG GAACCACCTA AATCACCAGT TCTGATACCT GCATCCAAAA
2821 CCTTTTTAAC TGCATCTTCA ATGGCCTTAC CTTCTTCAGG CAAGTTCAAT GACAATTTCA
2881 ACATCATTGC AGCAGACAAG ATAGTGGCGA TAGGGTTGAC CTTATTCTTT GGCAAATCTG
2941 GAGCAGAACC GTGGCATGGT TCGTACAAAC CAAATGCGGT GTTCTTGTCT GGCAAAGAGG
3001 CCAAGGACGC AGATGGCAAC AAACCCAAGG AACCTGGGAT AACGGAGGCT TCATCGGAGA
3061 TGATATCACC AAACATGTTG CTGGTGATTA TAATACCATT TAGGTGGGTT GGGTTCTTAA
3121 CTAGGATCAT GGCGGCAGAA TCAATCAATT GATGTTGAAC CTTCAATGTA GGAAATTCGT
3181 TCTTGATGGT TTCCTCCACA GTTTTTCTCC ATAATCTTGA AGAGGCCAAA ACATTAGCTT
3241 TATCCAAGGA CCAAATAGGC AATGGTGGCT CATGTTGTAG GGCCATGAAA GCGGCCATTC
3301 TTGTGATTCT TTGCACTTCT GGAACGGTGT ATTGTTCACT ATCCCAAGCG ACACCATCAC
3361 CATCGTCTTC CTTTCTCTTA CCAAAGTAAA TACCTCCCAC TAATTCTCTG ACAACAACGA
3421 AGTCAGTACC TTTAGCAAAT TGTGGCTTGA TTGGAGATAA GTCTAAAAGA GAGTCGGATG
3481 CAAAGTTACA TGGTCTTAAG TTGGCGTACA ATTGAAGTTC TTTACGGATT TTTAGTAAAC
3541 CTTGTTCAGG TCTAACACTA CCTGTACCCC ATTTAGGACC ACCCACAGCA CCTAACAAAA
3601 CGGCATCAAC CTTCTTGGAG GCTTCCAGCG CCTCATCTGG AAGTGGGACA CCTGTAGCGT
3661 CGATAGCAGC ACCACCAATT AAATGATTTT CGAAATCGAA CTTGACATTG GAACGAACAT
3721 CAGAAATAGC TTTAAGAACC TTAATGGCTT CGGCTGTGAT TTCTTGACCA ACGTGGTCAC
3781 CTGGCAAAAC GACGATCTTC TTAGGGGCAG ACATTAGAAT GGTATATCCT TGAAATATAT
3841 ATATATATTG CTGAAATGTA AAAGGTAAGA AAAGTTAGAA AGTAAGACGA TTGCTAACCA
3901 CCTATTGGAA AAAACAATAG GTCCTTAAAT AATATTGTCA ACTTCAAGTA TTGTGATGCA
3961 AGCATTTAGT CATGAACGCT TCTCTATTCT ATATGAAAAG CCGGTTCCGG CGCTCTCACC
4021 TTTCCTTTTT CTCCCAATTT TTCAGTTGAA AAAGGTATAT GCGTCAGGCG ACCTCTGAAA
4081 TTAACAAAAA ATTTCCAGTC ATCGAATTTG ATTCTGTGCG ATAGCGCCCC TGTGTGTTCT
4141 CGTTATGTTG AGGAAAAAAA TAATGGTTGC TAAGAGATTC GAACTCTTGC ATCTTACGAT
4201 ACCTGAGTAT TCCCACAGTT GGGGATCTCG ACTCTAGCTA GAGGATCAAT TCGTAATCAT
4261 GGTCATAGCT GTTTCCTGTG TGAAATTGTT ATCCGCTCAC AATTCCACAC AACATACGAG
4321 CCGGAAGCAT AAAGTGTAAA GCCTGGGGTG CCTAATGAGT GAGCTAACTC ACATTAATTG
4381 CGTTGCGCTC ACTGCCCGCT TTCCAGTCGG GAAACCTGTC GTGCCAGCTG ATAACTTCGT
4441 ATAATGTATG CTATACGAAG TTATTAGGTC TGAAGAGGAG TTTACGTCCA GCCAAGCTAG
4501 CTTGGCTGCA GGTCGAGCGG CCGCGATCCG GAACCCTTAA TATAACTTCG TATAATGTAT
4561 GCTATACGAA GTTATCAGCT GCATTAATGA ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG
4621 CGTATTGGGC GCTCTTCCGC TTCCTCGCTC ACTGACTCGC TGCGCTCGGT CGTTCGGCTG
4681 CGGCGAGCGG TATCAGCTCA CTCAAAGGCG GTAATACGGT TATCCACAGA ATCAGGGGAT
4741 AACGCAGGAA AGAACATGTG AGCAAAAGGC CAGCAAAAGG CCAGGAACCG TAAAAAGGCC
4801 GCGTTGCTGG CGTTTTTCCA TAGGCTCCGC CCCCCTGACG AGCATCACAA AAATCGACGC
4861 TCAAGTCAGA GGTGGCGAAA CCCGACAGGA CTATAAAGAT ACCAGGCGTT TCCCCCTGGA
4921 AGCTCCCTCG TGCGCTCTCC TGTTCCGACC CTGCCGCTTA CCGGATACCT GTCCGCCTTT
4981 CTCCCTTCGG GAAGCGTGGC GCTTTCTCAT AGCTCACGCT GTAGGTATCT CAGTTCGGTG
5041 TAGGTCGTTC GCTCCAAGCT GGGCTGTGTG CACGAACCCC CCGTTCAGCC CGACCGCTGC
5101 GCCTTATCCG GTAACTATCG TCTTGAGTCC AACCCGGTAA GACACGACTT ATCGCCACTG
5161 GCAGCAGCCA CTGGTAACAG GATTAGCAGA GCGAGGTATG TAGGCGGTGC TACAGAGTTC
5221 TTGAAGTGGT GGCCTAACTA CGGCTACACT AGAAGGACAG TATTTGGTAT CTGCGCTCTG
5281 CTGAAGCCAG TTACCTTCGG AAAAAGAGTT GGTAGCTCTT GATCCGGCAA ACAAACCACC
5341 GCTGGTAGCG GTGGTTTTTT TGTTTGCAAG CAGCAGATTA CGCGCAGAAA AAAAGGATCT
5401 CAAGAAGATC CTTTGATCTT TTCTACGGGG TCTGACGCTC AGTGGAACGA AAACTCACGT
5461 TAAGGGATTT TGGTCATGAG ATTATCAAAA AGGATCTTCA CCTAGATCCT TTTAAATTAA
5521 AAATGAAGTT TTAAATCAAT CTAAAGTATA TATGAGTAAA CTTGGTCTGA CAGTTACCAA
5581 TGCTTAATCA GTGAGGCACC TATCTCAGCG ATCTGTCTAT TTCGTTCATC CATAGTTGCC
5641 TGACTCCCCG TCGTGTAGAT AACTACGATA CGGGAGGGCT TACCATCTGG CCCCAGTGCT
5701 GCAATGATAC CGCGAGACCC ACGCTCACCG GCTCCAGATT TATCAGCAAT AAACCAGCCA
5761 GCCGGAAGGG CCGAGCGCAG AAGTGGTCCT GCAACTTTAT CCGCCTCCAT CCAGTCTATT
5821 AATTGTTGCC GGGAAGCTAG AGTAAGTAGT TCGCCAGTTA ATAGTTTGCG CAACGTTGTT
5881 GCCATTGCTA CAGGCATCGT GGTGTCACGC TCGTCGTTTG GTATGGCTTC ATTCAGCTCC
5941 GGTTCCCAAC GATCAAGGCG AGTTACATGA TCCCCCATGT TGTGCAAAAA AGCGGTTAGC
6001 TCCTTCGGTC CTCCGATCGT TGTCAGAAGT AAGTTGGCCG CAGTGTTATC ACTCATGGTT
6061 ATGGCAGCAC TGCATAATTC TCTTACTGTC ATGCCATCCG TAAGATGCTT TTCTGTGACT
6121 GGTGAGTACT CAACCAAGTC ATTCTGAGAA TAGTGTATGC GGCGACCGAG TTGCTCTTGC
6181 CCGGCGTCAA TACGGGATAA TACCGCGCCA CATAGCAGAA CTTTAAAAGT GCTCATCATT
6241 GGAAAACGTT CTTCGGGGCG AAAACTCTCA AGGATCTTAC CGCTGTTGAG ATCCAGTTCG
6301 ATGTAACCCA CTCGTGCACC CAACTGATCT TCAGCATCTT TTACTTTCAC CAGCGTTTCT
6361 GGGTGAGCAA AAACAGGAAG GCAAAATGCC GCAAAAAAGG GAATAAGGGC GACACGGAAA
6421 TGTTGAATAC TCATACTCTT CCTTTTTCAA TATTATTGAA GCATTTATCA GGGTTATTGT
6481 CTCATGAGCG GATACATATT TGAATGTATT TAGAAAAATA AACAAATAGG GGTTCCGCGC
6541 ACATTTCCCC GAAAAGTGCC ACCTGACGTC TAAGAAACCA TTATTATCAT GACATTAACC
6601 TATAAAAATA GGCGTATCAC GAGGCCCTTT CGTCTCGCGC GTTTCGGTGA TGACGGTGAA
6661 AACCTCTGAC ACATGCAGCT CCCGGAGACG GTCACAGCTT GTCTGTAAGC GGATGCCGGG
6721 AGCAGACAAG CCCGTCAGGG CGCGTCAGCG GGTGTTGGCG GGTGTCGGGG CTGGCTTAAC
6781 TATGCGGCAT CAGAGCAGAT TGTACTGAGA GTGCACCATA ACGCATTTAA GCATAAACAC
6841 GCACTATGCC GTTCTTCTCA TGTATATATA TATACAGGCA ACACGCAGAT ATAGGTGCGA
6901 CGTGAACAGT GAGCTGTATG TGCGCAGCTC GCGTTGCATT TTCGGAAGCG CTCGTTTTCG
6961 GAAACGCTTT GAAGTTCCTA TTCCGAAGTT CCTATTCTCT AGCTAGAAAG TATAGGAACT
7021 TCAGAGCGCT TTTGAAAACC AAAAGCGCTC TGAAGACGCA CTTTCAAAAA ACCAAAAACG
7081 CACCGGACTG TAACGAGCTA CTAAAATATT GCGAATACCG CTTCCACAAA CATTGCTCAA
7141 AAGTATCTCT TTGCTATATA TCTCTGTGCT ATATCCCTAT ATAACCTACC CATCCACCTT
7201 TCGCTCCTTG AACTTGCATC TAAACTCGAC CTCTACATTT TTTATGTTTA TCTCTAGTAT
7261 TACTCTTTAG ACAAAAAAAT TGTAGTAAGA ACTATTCATA GAGTGAATCG AAAACAATAC
7321 GAAAATGTAA ACATTTCCTA TACGTAGTAT ATAGAGACAA AATAGAAGAA ACCGTTCATA
7381 ATTTTCTGAC CAATGAAGAA TCATCAACGC TATCACTTTC TGTTCACAAA GTATGCGCAA
7441 TCCACATCGG TATAGAATAT AATCGGGGAT GCCTTTATCT TGAAAAAATG CACCCGCAGC
7501 TTCGCTAGTA ATCAGTAAAC GCGGGAAGTG GAGTCAGGCT TTTTTTATGG AAGAGAAAAT
7561 AGACACCAAA GTAGCCTTCT TCTAACCTTA ACGGACCTAC AGTGCAAAAA GTTATCAAGA
7621 GACTGCATTA TAGAGCGCAC AAAGGAGAAA AAAAGTAATC TAAGATGCTT TGTTAGAAAA
7681 ATAGCGCTCT CGGGATGCAT TTTTGTAGAA CAAAAAAGAA GTATAGATTC TTTGTTGGTA
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