LOCUS Exported 5741 bp ds-DNA circular SYN 21-DEC-2017
DEFINITION Yeast episomal plasmid with a LEU2 marker.
COMMENT The sequence of this plasmid was reconstructed using the description
from Gietz and Sugino.
FEATURES Location/Qualifiers
source 1..5741
/organism="synthetic DNA construct"
/lab_host="Saccharomyces cerevisiae"
/mol_type="other DNA"
promoter 142..172
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 180..196
/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)."
CDS 216..500
/codon_start=1
/gene="lacZ"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRI"
misc_feature 233..289
/label=MCS
/note="pUC19 multiple cloning site"
rep_origin 745..2091
/label=2u ori
/note="yeast 2u plasmid origin of replication"
CDS complement(2227..3321)
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/label=LEU2
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
promoter complement(3322..3729)
/gene="S. cerevisiae LEU2"
/label=LEU2 promoter
promoter 3835..3939
/gene="bla"
/label=AmpR promoter
CDS 3940..4800
/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 4971..5559
/direction=RIGHT
/label=ori
/note="high-copy-number colE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
241 atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg
301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
421 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
481 gcggtatttc acaccgcata tatcggatcg tacttgttac ccatcattga attttgaaca
541 tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat aataatatat
601 agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa ctattgcatc
661 tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt tccatcttgc
721 acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag aacaaaaatg
781 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
841 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
901 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
961 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
1021 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
1081 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
1141 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
1201 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
1261 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
1321 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
1381 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
1441 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
1501 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
1561 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
1621 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag
1681 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
1741 ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag
1801 ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat
1861 atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc
1921 tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg
1981 ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa
2041 ttggattagt ctcatccttc aatgctatca tttcctttga tattggatcg atccgatgat
2101 aagctgtcaa acatgagaat taattctacc ctatgaacat attccatttt gtaatttcgt
2161 gtcgtttcta ttatgaattt catttataaa gtttatgtac aaatatcata aaaaaagaga
2221 atctttttaa gcaaggattt tcttaacttc ttcggcgaca gcatcaccga cttccgtggt
2281 actgttggaa ccacctaaat caccagttct gatacctgca tccaaaacct ttttaactgc
2341 atcttcaatg gccttacctt cttcaggcaa gttcaatgac aatttcaaca tcattgcagc
2401 agacaagata gtggcgatag ggttgacctt attctttggc aaatctggag cagaaccgtg
2461 gcatggttcg tacaaaccaa atgcggtgtt cttgtctggc aaagaggcca aggacgcaga
2521 tggcaacaaa cccaaggaac ctgggataac ggaggcttca tcggagatga tatcaccaaa
2581 catgttgctg gtgattataa taccatttag gtgggttggg ttcttaacta ggatcatggc
2641 ggcagaatca atcaattgat gttgaacctt caatgtagga aattcgttct tgatggtttc
2701 ctccacagtt tttctccata atcttgaaga ggccaaaaca ttagctttat ccaaggacca
2761 aataggcaat ggtggctcat gttgtagggc catgaaagcg gccattcttg tgattctttg
2821 cacttctgga acggtgtatt gttcactatc ccaagcgaca ccatcaccat cgtcttcctt
2881 tctcttacca aagtaaatac ctcccactaa ttctctgaca acaacgaagt cagtaccttt
2941 agcaaattgt ggcttgattg gagataagtc taaaagagag tcggatgcaa agttacatgg
3001 tcttaagttg gcgtacaatt gaagttcttt acggattttt agtaaacctt gttcaggtct
3061 aacactacct gtaccccatt taggaccacc cacagcacct aacaaaacgg catcaacctt
3121 cttggaggct tccagcgcct catctggaag tgggacacct gtagcatcga tagcagcacc
3181 accaattaaa tgattttcga aatcgaactt gacattggaa cgaacatcag aaatagcttt
3241 aagaacctta atggcttcgg ctgtgatttc ttgaccaacg tggtcacctg gcaaaacgac
3301 gatcttctta ggggcagaca ttagaatggt atatccttga aatatatata tatatattgc
3361 tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat tgctaaccac ctattggaaa
3421 aaacaatagg tccttaaata atattgtcaa cttcaagtat tgtgatgcaa gcatttagtc
3481 atgaacgctt ctctattcta tatgaaaagc cggttccgcg gctctcacct ttcctttttc
3541 tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat taacaaaaaa
3601 tttccagtca tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc gttatgttga
3661 ggaaaaaaat aatggttgct aagagattcg aactcttgca tcttacgata cctgagtatt
3721 cccacagtta attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
3781 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
3841 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
3901 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
3961 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
4021 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
4081 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
4141 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
4201 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
4261 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
4321 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
4381 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
4441 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
4501 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
4561 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
4621 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
4681 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
4741 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
4801 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
4861 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
4921 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
4981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
5041 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
5101 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
5161 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
5221 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
5281 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
5341 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
5401 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
5461 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
5521 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
5581 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
5641 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
5701 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag a
//