LOCUS       Exported                8182 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRS4213, complete sequence.
ACCESSION   AF174133
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8182)
  AUTHORS   Tomlin GC, Wixon JL, Bolotin-Fukuhara M, Oliver SG.
  TITLE     A new family of yeast vectors and S288C-derived strains for the 
            systematic analysis of gene function
  JOURNAL   Yeast 18 (6), 563-575 (2001)
  PUBMED    11284012
REFERENCE   2  (bases 1 to 8182)
  AUTHORS   Tomlin GC, Wixon JL, Bolotin-Fukuhara M, Oliver SG.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-AUG-1999) Biological Sciences, University of 
            Manchester, 2.205 Stopford Building, Oxford Rd, Manchester M13 9PT, 
            United Kingdom
REFERENCE   3  (bases 1 to 8182)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8182
                     /organism="Cloning vector pRS4213"
                     /mol_type="genomic DNA"
                     /label=yeast episomal vector with ADE8 and HIS3 markers
                     /note="yeast episomal vector with ADE8 and HIS3 markers"
                     /db_xref="taxon:102653"
     misc_feature    339..2237
                     /label=ADE8 gene cassette
                     /note="ADE8 gene cassette"
     CDS             1195..1839
                     /codon_start=1
                     /product="ADE8 protein"
                     /label=ADE8 protein
                     /protein_id="AAD54287.1"
                     /translation="MARIVVLISGSGSNLQALIDAQKQGQLGEDAHIVSVISSSKKAYG
                     LTRAADNNIPTKVCSLYPYTKGIAKEDKAARAKARSQFENDLAKLVLEEKPDVIICAGW
                     LLILGSTFLSQLQSVPILNLHPALPGCFDGTTHAIEMAWRKCQDENKPLTAGCMVHYVI
                     EEVDKGEPLVVKKLEIIPGEETLEQYEQRVHDAEHIAIVEATYKVLQQLHK"
     misc_feature    2238..3250
                     /label=HIS3 gene cassette
                     /note="HIS3 gene cassette"
     promoter        2246..2433
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             2434..3096
                     /codon_start=1
                     /product="HIS3 protein"
                     /label=HIS3 protein
                     /protein_id="AAD54288.1"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(3811..4266)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(4243..4623)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     4407..4423
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4433..4451
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4460..4567
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     misc_feature    4464..4566
                     /label=polycloning site
                     /note="polycloning site"
     primer_bind     4477..4493
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4527..4543)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4580..4598)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4619..4635)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4643..4659
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(4667..4697)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(5021..5609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5780..6640)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6641..6745)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6772..8114
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(7735..7782)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."