LOCUS       40924_37898        4788 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast centromere vector pRS414 with TRP1 marker, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4788)
  AUTHORS   Sikorski RS, Hieter P.
  TITLE     A system of shuttle vectors and yeast host strains designed for 
            efficient manipulation of DNA in Saccharomyces cerevisiae
  JOURNAL   Genetics 122 (1), 19-27 (1989)
  PUBMED    2659436
REFERENCE   2  (bases 1 to 4788)
  AUTHORS   Christianson TW, Sikorski RS, Dante M, Shero JH, Hieter P.
  TITLE     Multifunctional yeast high-copy-number shuttle vectors
  JOURNAL   Gene 110 (1), 119-122 (1992)
  PUBMED    1544568
REFERENCE   3  (bases 1 to 4788)
  AUTHORS   Stillman DJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-NOV-1993) David J. Stillman, Dept. of Cellular, Viral 
            and Molecular Biology, University of Utah Medical Center, Salt Lake 
            City, UT 84132 USA
REFERENCE   4  (bases 1 to 4788)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4788)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics"; 
            date: "1989"; volume: "122"; issue: "1"; pages: "19-27"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Gene"; 
            date: "1992"; volume: "110"; issue: "1"; pages: "119-122"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (11-NOV-1993) David J. Stillman, Dept. of Cellular, Viral and 
            Molecular Biology, University of Utah Medical Center, Salt Lake 
            City, UT 84132 USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        187..467
                     /label=TRP1 promoter
     CDS             468..1139
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1241..1696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1841..1857
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1867..1885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(1894..2001)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(2014..2032)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2053..2069)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2077..2093)
                     /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(2101..2131)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2146..2167)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2455..3043)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3217..4074)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4075..4179)
                     /label=AmpR promoter
     misc_feature    4216..4719
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat
      241 atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
      301 aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
      361 gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
      421 tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
      481 atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
      541 cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
      601 agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
      661 atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
      721 ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
      781 ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
      841 actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
      901 attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
      961 gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     1021 atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     1081 agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     1141 aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgccta tgcggtgtga
     1201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     1261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1681 ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1741 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     1801 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     1861 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac
     1921 ggtatcgata agcttgatat cgaattcctg cagcccgggg gatccactag ttctagagcg
     1981 gccgccaccg cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     2041 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2101 caacatagga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgaggtaact
     2161 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2221 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2281 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2341 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2401 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2461 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2521 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2581 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2641 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2701 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2761 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2821 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2881 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2941 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3001 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3061 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3121 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3181 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3241 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3301 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3361 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3421 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3481 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3541 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3601 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3661 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3721 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3781 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3841 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3901 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3961 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4021 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4081 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4141 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4201 acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa
     4261 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt
     4321 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt
     4381 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg
     4441 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc
     4501 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt
     4561 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat
     4621 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt
     4681 aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcctaagaa accattatta
     4741 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
