LOCUS       .                       7621 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     rep_origin      6200..7080
                     /note="yeast 2μ plasmid origin of replication"
                     /note="color: #ffff00"
                     /label="2μ ori"
     CDS             complement(3621..4481)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="color: #ccffcc"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     CDS             complement(4577..5380)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /note="yeast auxotrophic marker, counterselectable with
                     5-fluoroorotic acid (5-FOA)"
                     /note="color: #ff7f50"
                     /product="orotidine-5'-phosphate decarboxylase, required
                     for uracil biosynthesis"
                     /transl_table=1
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN*"
                     /label="URA3"
     CDS             771..1430
                     /codon_start=1
                     /gene="cat"
                     /note="confers resistance to chloramphenicol"
                     /note="color: #ccffcc"
                     /product="chloramphenicol acetyltransferase"
                     /transl_table=1
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA*"
                     /label="CmR"
     rep_origin      complement(2862..3450)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /note="color: #ffff00"
                     /label="ori"
     rep_origin      complement(7149..7604)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
                     /label="f1 ori"
     promoter        2..443
                     /gene="S. cerevisiae GAL1"
                     /note="inducible promoter, regulated by Gal4"
                     /note="color: #ffffff"
                     /label="GAL1 promoter"
     CDS             1772..2077
                     /codon_start=1
                     /gene="ccdB"
                     /note="Plasmids containing the ccdB gene cannot be
                     propagated in standard E. coli strains."
                     /note="color: #993366"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /transl_table=1
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI*"
                     /label="ccdB"
     terminator      2367..2614
                     /gene="S. cerevisiae CYC1"
                     /note="transcription terminator for CYC1"
                     /note="color: #ffffff"
                     /label="CYC1 terminator"
     promoter        complement(5381..5596)
                     /gene="S. cerevisiae URA3"
                     /label="URA3 promoter"
                     /note="color: #ffffff"
     protein_bind    511..635
                     /bound_moiety="LR Clonase™"
                     /gene="mutant version of attR"
                     /note="recombination site for the Gateway® LR reaction"
                     /note="color: #31849b"
                     /label="attR1"
     protein_bind    complement(2118..2242)
                     /bound_moiety="LR Clonase™"
                     /gene="mutant version of attR"
                     /note="recombination site for the Gateway® LR reaction"
                     /note="color: #31849b"
                     /label="attR2"
     protein_bind    2..119
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating
                     Gal4-dependent induction"
                     /note="color: #31849b"
                     /label="UAS"
     CDS             2265..2306
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
                     /note="color: #cc99b2"
     promoter        687..717
                     /note="E. coli lac promoter with an 'up' mutation"
                     /note="color: #ffffff"
                     /label="lac UV5 promoter"
     promoter        475..493
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff"
                     /label="T7 promoter"
     CDS             2316..2333
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
ORIGIN
        1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
       61 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
      121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
      181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
      241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
      301 taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc
      361 ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac
      421 ctctatactt taacgtcaag gagaaaaaac cccggatcgg actactagca gctgtaatac
      481 gactcactat agggaatatt aagctatcaa acaagtttgt acaaaaaagc tgaacgagaa
      541 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
      601 taatactgta aaacacaaca tatccagtca ctatggcggc cgcgggtgat gctgccaact
      661 tagcggccgc attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt
      721 ggattttgag ttaggatccg tcgagatttt caggagctaa ggaagctaaa atggagaaaa
      781 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
      841 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
      901 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
      961 cccgcctgat gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga
     1021 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     1081 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     1141 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     1201 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     1261 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
     1321 tgatgccgct ggcgattcag gttcatcatg ccgtctgtga tggcttccat gtcggcagaa
     1381 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tctagaggat
     1441 ccggcttact aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa
     1501 gaatatatac tgatatgtat acccgaagta tgtcaaaaag aggtgtgcta tgaagcagcg
     1561 tattacagtg acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat
     1621 atctccggtc tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc
     1681 tggaaagcgg aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc
     1741 tcttttgctg acgagaacag ggactggtga aatgcagttt aaggtttaca cctataaaag
     1801 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg
     1861 acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact
     1921 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
     1981 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
     2041 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccg ttatacacag
     2101 ccagtctgca ggtcgaccat agtgactgga tatgttgtgt tttacagtat tatgtagtct
     2161 gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt
     2221 cagctttctt gtacaaagtg gttcgatcta gagggccctt cgaaggtaag cctatcccta
     2281 accctctcct cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt
     2341 aaacccgctg atcctagagg gccgcatcat gtaattagtt atgtcacgct tacattcacg
     2401 ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     2461 ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
     2521 tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga
     2581 gaaggttttg ggacgctcga aggctttaat ttgcaagctg cggccctgca ttaatgaatc
     2641 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2701 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2761 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2821 caaaagccca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2881 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2941 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3001 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3061 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3121 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3181 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3241 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3301 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3361 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3421 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3481 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3541 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3601 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3661 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3721 gagcgcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3781 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3841 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3901 ccagttaata gtttgcgcaa cgttgttggc attgctacag gcatcgtggt gtcactctcg
     3961 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4021 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4081 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4141 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4201 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatag tgtatcacat
     4261 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4321 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4381 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4441 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatgg
     4501 gtaataactg atataattaa attgaagctc taatttgtga gtttagtata catgcattta
     4561 cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct tcccagcctg
     4621 cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat agtcctcttc
     4681 caacaataat aatgtcagat cctgtagaga ccacatcatc cacggttcta tactgttgac
     4741 ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt cataatcaac caatcgtaac
     4801 cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa tctttgtcgc
     4861 tcttcgcaat gtcaacagta cccttagtat attctccagt agatagggag cccttgcatg
     4921 acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttct gccgcctgct
     4981 tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg tctgcccatt
     5041 ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca atgtcagcaa
     5101 attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt agcggcttaa
     5161 ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt aaacaaattt
     5221 tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca tccaatgaag
     5281 cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca acaggactag
     5341 gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt cgtttcctgc
     5401 aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt tcttcaacac
     5461 tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt
     5521 cggagattac cgaatcaaaa aaatttcaaa gaaaccgaaa tcaaaaaaaa gaataaaaaa
     5581 aaaatgatga attgaattga aaagctagct tatcgatgat aagctgtcaa agatgagaat
     5641 taattccacg gactatagac tatactagat actccgtcta ctgtacgata cacttccgct
     5701 caggtccttg tcctttaacg aggccttacc actcttttgt tactctattg atccagctca
     5761 gcaaaggcag tgtgatctaa gattctatct tcgcgatgta gtaaaactag ctagaccgag
     5821 aaagagacta gaaatgcaaa aggcacttct acaatggctg ccatcattat tatccgatgt
     5881 gacgctgcag cttctcaatg atattcgaat acgctttgag gagatacagc ctaatatccg
     5941 acaaactgtt ttacagattt acgatcgtac ttgttaccca tcattgaatt ttgaacatcc
     6001 gaacctggga gttttccctg aaacagatag tatatttgaa cctgtataat aatatatagt
     6061 ctagcgcttt acggaagaca atgtatgtat ttcggttcct ggagaaacta ttgcatctat
     6121 tgcataggta atcttgcacg tcgcatcccc ggttcatttt ctgcgtttcc atcttgcact
     6181 tcaatagcat atctttgtta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa
     6241 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     6301 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
     6361 tgcaacgcga cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     6421 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     6481 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     6541 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     6601 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     6661 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     6721 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     6781 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     6841 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     6901 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     6961 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     7021 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     7081 gggaagctcc accccggttg ataatcagaa aagccccaaa aacaggaaga ttgtataagc
     7141 aaatatttaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     7201 cagctcattt tttaacgaat agcccgaaat cggcaaaatc ccttataaat caaaagaata
     7261 gaccgagata gggttgagtg ttgttccagt ttccaacaag agtccactat taaagaacgt
     7321 ggactccaac gtcaaagggc gaaaaagggt ctatcagggc gatggcccac tacgtgaacc
     7381 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcagtaaatc ggaagggtaa
     7441 acggatgccc ccatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     7501 gaagaaagcg aaaggagcgg gggctagggc ggtgggaagt gtaggggtca cgctgggcgt
     7561 aaccaccaca cccgccgcgc ttaatggggc gctacagggc gcgtggggat gatccactag
     7621 t
//
