LOCUS       Exported                5625 bp DNA     circular SYN 20-JUL-2025
DEFINITION  Yeast episomal vector with a TRP1 marker and an MCS derived from 
            pBLUESCRIPT II.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5625)
  AUTHORS   Christianson TW, Sikorski RS, Dante M, Shero JH, Hieter P.
  TITLE     Multifunctional yeast high-copy-number shuttle vectors.
  JOURNAL   Gene 1992;110:119-22.
  PUBMED    1544568
REFERENCE   2  (bases 1 to 5625)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5625)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5625)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5625)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1992"; volume: "110"; pages: "119-22"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1194..1197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        191..472
                     /label=TRP1 promoter
     CDS             473..1144
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1250..1705)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1850..1866
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1876..1894
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(1903..2010)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(2023..2041)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2062..2078)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2086..2102)
                     /label=lac operator
                     /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)."
     promoter        complement(2110..2140)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2155..2176)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2464..3052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3226..4083)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4084..4188)
                     /label=AmpR promoter
     rep_origin      complement(4215..5557)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatagatc aacgacatta ctatatatat aatataggaa gcatttaata gaacagcatc
      241 gtaatatatg tgtactttgc agttatgacg ccagatggca gtagtggaag atattcttta
      301 ttgaaaaata gcttgtcacc ttacgtacaa tcttgatccg gagcttttct ttttttgccg
      361 attaagaatt aattcggtcg aaaaaagaaa aggagagggc caagagggag ggcattggtg
      421 actattgagc acgtgagtat acgtgattaa gcacacaaag gcagcttgga gtatgtctgt
      481 tattaatttc acaggtagtt ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga
      541 ggccgcagaa tgtgctctag attccgatgc tgacttgctg ggtattatat gtgtgcccaa
      601 tagaaagaga acaattgacc cggttattgc aaggaaaatt tcaagtcttg taaaagcata
      661 taaaaatagt tcaggcactc cgaaatactt ggttggcgtg tttcgtaatc aacctaagga
      721 ggatgttttg gctctggtca atgattacgg cattgatatc gtccaactgc atggagatga
      781 gtcgtggcaa gaataccaag agttcctcgg tttgccagtt attaaaagac tcgtatttcc
      841 aaaagactgc aacatactac tcagtgcagc ttcacagaaa cctcattcgt ttattccctt
      901 gtttgattca gaagcaggtg ggacaggtga acttttggat tggaactcga tttctgactg
      961 ggttggaagg caagagagcc ccgaaagctt acattttatg ttagctggtg gactgacgcc
     1021 agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt ggtgttgatg taagcggagg
     1081 tgtggagaca aatggtgtaa aagactctaa caaaatagca aatttcgtca aaaatgctaa
     1141 gaaataggtt attactgagt agtatttatt taagtattgt ttgtgcactt gccgatctat
     1201 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     1261 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     1321 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     1381 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     1441 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     1501 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     1561 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     1621 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     1681 cttaatgcgc cgctacaggg cgcgtcgcgc cattcgccat tcaggctgcg caactgttgg
     1741 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     1801 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     1861 gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg gccccccctc
     1921 gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccggggg atccactagt
     1981 tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     2041 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2101 aattccacac aacataggag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2161 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2221 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2281 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2341 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2401 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2461 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2521 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2581 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2641 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2701 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2761 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2821 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2881 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2941 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3001 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3061 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3121 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3181 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3241 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3301 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3361 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3421 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3481 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3541 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3601 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3661 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3721 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3781 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3841 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3901 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3961 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4021 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4081 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4141 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4201 aaaagtgcca cctgaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga
     4261 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     4321 cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa
     4381 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     4441 caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa
     4501 tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg
     4561 tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa
     4621 gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg
     4681 tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta
     4741 tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat
     4801 tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat
     4861 aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa
     4921 tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag
     4981 atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat
     5041 atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt
     5101 agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg
     5161 gttttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga
     5221 acttcaaagc gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac
     5281 agctcactgt tcacgtcgca cctatatctg cgtgttgcct gtatatatat atacatgaga
     5341 agaacggcat agtgcgtgtt tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg
     5401 atgaaaggta gtctagtacc tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc
     5461 ttccttcagc actacccttt agctgttcta tatgctgcca ctcctcaatt ggattagtct
     5521 catccttcaa tgctatcatt tcctttgata ttggatcata ctaagaaacc attattatca
     5581 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
