LOCUS       Exported                4784 bp DNA     circular SYN 24-OCT-2024
DEFINITION  Yeast centromere vector with a TRP1 marker and an MCS derived from 
            pBLUESCRIPT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4784)
  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 1989;122:19-27.
  PUBMED    2659436
REFERENCE   2  (bases 1 to 4784)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4784)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4784)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics"; 
            date: "1989"; volume: "122"; pages: "19-27"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        187..468
                     /label=TRP1 promoter
     CDS             469..1140
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1240..1695)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1840..1856
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1863..1881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1890..1997
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(2010..2028)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2049..2065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2073..2089)
                     /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(2097..2127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2142..2163)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2451..3039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3213..4070)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4071..4175)
                     /label=AmpR promoter
     misc_feature    4212..4715
                     /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 ttaatagaac agcatcgtaa
      241 tatatgtgta ctttgcagtt atgacgccag atggcagtag tggaagatat tctttattga
      301 aaaatagctt gtcaccttac gtacaatctt gatccggagc ttttcttttt ttgccgatta
      361 agaattaatt cggtcgaaaa aagaaaagga gagggccaag agggagggca ttggtgacta
      421 ttgagcacgt gagtatacgt gattaagcac acaaaggcag cttggagtat gtctgttatt
      481 aatttcacag gtagttctgg tccattggtg aaagtttgcg gcttgcagag cacagaggcc
      541 gcagaatgtg ctctagattc cgatgctgac ttgctgggta ttatatgtgt gcccaataga
      601 aagagaacaa ttgacccggt tattgcaagg aaaatttcaa gtcttgtaaa agcatataaa
      661 aatagttcag gcactccgaa atacttggtt ggcgtgtttc gtaatcaacc taaggaggat
      721 gttttggctc tggtcaatga ttacggcatt gatatcgtcc aactgcatgg agatgagtcg
      781 tggcaagaat accaagagtt cctcggtttg ccagttatta aaagactcgt atttccaaaa
      841 gactgcaaca tactactcag tgcagcttca cagaaacctc attcgtttat tcccttgttt
      901 gattcagaag caggtgggac aggtgaactt ttggattgga actcgatttc tgactgggtt
      961 ggaaggcaag agagccccga aagcttacat tttatgttag ctggtggact gacgccagaa
     1021 aatgttggtg atgcgcttag attaaatggc gttattggtg ttgatgtaag cggaggtgtg
     1081 gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt tcgtcaaaaa tgctaagaaa
     1141 taggttatta ctgagtagta tttatttaag tattgtttgt gcacttgcct gcggtgtgaa
     1201 ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaaac gttaatattt
     1261 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     1321 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     1381 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     1441 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     1501 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     1561 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     1621 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     1681 cgctacaggg cgcgtcgcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     1741 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaggg gggatgtgct gcaaggcgat
     1801 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     1861 tgtaatacga ctcactatag ggcgaattgg agctccaccg cggtggcggc cgctctagaa
     1921 ctagtggatc ccccgggctg caggaattcg atatcaagct tatcgatacc gtcgacctcg
     1981 agggggggcc cggtacccag cttttgttcc ctttagtgag ggttaattcc gagcttggcg
     2041 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2101 ataggagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag gtaactcaca
     2161 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2221 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2281 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2461 ctcggccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2521 acaggactat aaagatacca ggcgttcccc cctggaagct ccctcgtgcg ctctcctgtt
     2581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2641 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2881 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3001 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3181 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3241 tcagcgatct gtctatttcg ttcatccata gttgcctgac tgcccgtcgt gtagataact
     3301 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3361 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3421 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3481 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3541 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3601 acatgatccc ccatgttgtg aaaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3661 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3721 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3781 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3841 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3901 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3961 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4021 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4081 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4141 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4201 gggtcctttt catcacgtgc tataaaaata attataattt aaatttttta atataaatat
     4261 ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt tgttttccga
     4321 agatgtaaaa gactctaggg ggatcgccaa caaatactac cttttatctt gctcttcctg
     4381 ctctcaggta ttaatgccga attgtttcat cttgtctgtg tagaagacca cacacgaaaa
     4441 tcctgtgatt ttacatttta cttatcgtta atcgaatgta tatctattta atctgctttt
     4501 cttgtctaat aaatatatat gtaaagtacg ctttttgttg aaatttttta aacctttgtt
     4561 tatttttttt tcttcattcc gtaactcttc taccttcttt atttactttc taaaatccaa
     4621 atacaaaaca taaaaataaa taaacacaga gtaaattccc aaattattcc atcattaaaa
     4681 gatacgaggc gcgtgtaagt tacaggcaag cgatccgtcc taagaaacca ttattatcat
     4741 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
