LOCUS       40924_48048        5825 bp DNA     circular SYN 01-JAN-1980
DEFINITION  High-copy episomal Saccharomyces cerevisiae expression vector that 
            serves as an acceptor in the Echo(TM) Cloning System.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5825)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5825)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..443
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     promoter        475..493
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(507..540)
                     /label=loxH
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021). The loxH variant 
                     was optimized for expression constructs but remains 
                     compatible with loxP."
     terminator      571..818
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(1066..1654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2123..2501)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene;
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     CDS             complement(2784..3584)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(3585..3805)
                     /label=URA3 promoter
     rep_origin      complement(4404..5284)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(5295..5808)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
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 aagcttatta cctcatatag catacattat acgaagttat
      541 gtttaaaccc gctgatccta gagggccgca tcatgtaatt agttatgtca cgcttacatt
      601 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
      661 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
      721 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
      781 ttgagaaggt tttgggacgc tcgaaggctt taatttgcaa gctgcggccc tgcattaatg
      841 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
      901 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      961 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1021 ccagcaaaag cccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1081 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1141 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1201 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1261 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1321 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1381 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1441 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1501 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1561 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1621 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1681 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1741 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1801 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1861 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1921 acgggagcgc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1981 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2041 tgcaacttta tccgcctcca ttcagtctat taattgttgc cgggaagcta gagtaagtag
     2101 ttcgccagtt aatagtttgc gcaacgttgt tggcattgct acaggcatcg tggtgtcact
     2161 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2221 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2281 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2341 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2401 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata atagtgtatc
     2461 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2521 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2581 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2641 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2701 atgggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag tatacatgca
     2761 tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata tgcttcccag
     2821 cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc aaatagtcct
     2881 cttccaacaa taataatgtc agatcctgta gagaccacat catccacggt tctatactgt
     2941 tgacccaatg cgtctccctt gtcatctaaa cccacaccgg gtgtcataat caaccaatcg
     3001 taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac aaaatctttg
     3061 tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag ggagcccttg
     3121 catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc ttctgccgcc
     3181 tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt aatgtctgcc
     3241 cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt accaatgtca
     3301 gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc
     3361 ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt tagtaaacaa
     3421 attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg aacatccaat
     3481 gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc agcaacagga
     3541 ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta tcttcgtttc
     3601 ctgcaggttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca tgtttcttca
     3661 acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg ttcttccttc
     3721 tgttcggaga ttaccgaatc aaaaaaattt caaagaaacc gaaatcaaaa aaaagaataa
     3781 aaaaaaaatg atgaattgaa ttgaaaagct agcttatcga tgataagctg tcaaagatga
     3841 gaattaattc cacggactat agactatact agatactccg tctactgtac gatacacttc
     3901 cgctcaggtc cttgtccttt aacgaggcct taccactctt ttgttactct attgatccag
     3961 ctcagcaaag gcagtgtgat ctaagattct atcttcgcga tgtagtaaaa ctagctagac
     4021 cgagaaagag actagaaatg caaaaggcac ttctacaatg gctgccatca ttattatccg
     4081 atgtgacgct gcagcttctc aatgatattc gaatacgctt tgaggagata cagcctaata
     4141 tccgacaaac tgttttacag atttacgatc gtacttgtta cccatcattg aattttgaac
     4201 atccgaacct gggagttttc cctgaaacag atagtatatt tgaacctgta taataatata
     4261 tagtctagcg ctttacggaa gacaatgtat gtatttcggt tcctggagaa actattgcat
     4321 ctattgcata ggtaatcttg cacgtcgcat ccccggttca ttttctgcgt ttccatcttg
     4381 cacttcaata gcatatcttt gttaacgaag catctgtgct tcattttgta gaacaaaaat
     4441 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     4501 aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca
     4561 aaaatgcaac gcgacgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     4621 agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt
     4681 gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt
     4741 ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg
     4801 ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact
     4861 ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg
     4921 catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat
     4981 agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta
     5041 tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag
     5101 ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga
     5161 ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata
     5221 tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg
     5281 caatgggaag ctccaccccg gttgataatc agaaaagccc caaaaacagg aagattgtat
     5341 aagcaaatat ttaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt
     5401 aaatcagctc attttttaac gaatagcccg aaatcggcaa aatcccttat aaatcaaaag
     5461 aatagaccga gatagggttg agtgttgttc cagtttccaa caagagtcca ctattaaaga
     5521 acgtggactc caacgtcaaa gggcgaaaaa gggtctatca gggcgatggc ccactacgtg
     5581 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcagta aatcggaagg
     5641 gtaaacggat gcccccattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5701 aagggaagaa agcgaaagga gcgggggcta gggcggtggg aagtgtaggg gtcacgctgg
     5761 gcgtaaccac cacacccgcc gcgcttaatg gggcgctaca gggcgcgtgg ggatgatcca
     5821 ctagt
//
