LOCUS       Exported                9560 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRS327, complete sequence.
ACCESSION   AY150810
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9560)
  AUTHORS   Eriksson P, Thomas LR, Thorburn A, Stillman DJ.
  TITLE     pRS yeast vectors with a LYS2 marker
  JOURNAL   BioTechniques 36 (2), 212-213 (2004)
  PUBMED    14989082
REFERENCE   2  (bases 1 to 9560)
  AUTHORS   Thomas LR, Eriksson P, Stillman DJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-SEP-2002) Pathology, University of Utah, 30 North 1900
            East Rm 5C126 SOM, Salt Lake City, UT 84132-2501, USA
REFERENCE   3  (bases 1 to 9560)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9560
                     /organism="Cloning vector pRS327"
                     /mol_type="genomic DNA"
                     /note="YEp vector; Saccharomyces cerevisiae/Escherichia 
                     coli shuttle vector"
                     /db_xref="taxon:209532"
     CDS             complement(717..4895)
                     /codon_start=1
                     /gene="LYS2"
                     /product="Lys2"
                     /label=LYS2
                     /protein_id="AAN62755.1"
                     /translation="MTNEKVWIEKLDNPTLSVLPHDFLRPQQEPYTKQATYSLQLPQLD
                     VPHDSFSNKYAVALSVWAALIYRVTGDDDIVLYIANNKILRFNIQPTWSFNELYSTINN
                     ELNKLNSIEANFSFDELAEKIQSCQDLERTPQLFRLAFLENQDFKLDEFKHHLVDFALN
                     LDTSNNAHVLNLIYNSLLYSNERVTIVADQFTQYLTAALSDPSNCITKISLITASSKDS
                     LPDPTKNLGWCDFVGCIHDIFQDNAEAFPERTCVVETPTLNSDKSRSFTYRDINRTSNI
                     VAHYLIKTGIKRGDVVMIYSSRGVDLMVCVMGVLKAGATFSVIDPAYPPARQTIYLGVA
                     KPRGLIVIRAAGQLDQLVEDYINDELEIVSRINSIAIQENGTIEGGKLDNGEDVLAPYD
                     HYKDTRTGVVVGPDSNPTLSFTSGSEGIPKGVLGRHFSLAYYFNWMSKRFNLTENDKFT
                     MLSGIAHDPIQRDMFTPLFLGAQLYVPTQDDIGTPGRLAEWMSKYGCTVTHLTPAMGQL
                     LTAQATTPFPKLHHAFFVGDILTKRDCLRLQTLAENCRIVNMYGTTETQRAVSYFEVKS
                     KNDDPNFLKKLKDVMPAGKGMLNVQLLVVNRNDRTQICGIGEIGEIYVRAGGLAEGYRG
                     LPELNKEKFVNNWFVEKDHWNYLDKDNGEPWRQFWLGPRDRLYRTGDLGRYLPNGDCEC
                     CGRADDQVKIRGFRIELGEIDTHISQHPLVRENITLVRKNADNEPTLITFMVPRFDKPD
                     DLSKFQSDVPKEVETDPIVKGLIGYHLLSKDIRTFLKKRLASYAMPSLIVVMDKLPLNP
                     NGKVDKPKLQFPTPKQLNLVAENTVSETDDSQFTNVEREVRDLWLSILPTKPASVSPDD
                     SFFDLGGHSILATKMIFTLKKKLQVDLPLGTIFKYPTIKAFAAEIDRIKSSGGSSQGEV
                     VENVTANYAEDAKKLVETLPSSYPSREYFVEPNSAEGKTTINVFVTGVTGFLGSYILAD
                     LLGRSPKNYSFKVFAHVRAKDEEAAFARLQKAGITYGTWNEKFASNIKVVLGDLSKSQF
                     GLSDEKWMDLANTVDIIIHNGALVHWVYPYAKLRDPNVISTINVMSLAAVGKPKFFDFV
                     SSTSTLDTEYYFNLSDKLVSEGKPGILESDDLMNSASGLTGGYGQSKWAAEYIIRRAGE
                     RGLRGCIVRPGYVTGASANGSSNTDDFLLRFLKGSVQLGKIPDIENSVNMVPVDHVARV
                     VVATSLNPPKENELAVAQVTGHPRILFKDYLYTLHDYGYDVEIESYSKWKKSLEASVID
                     RNEENALYPLLHMVLDNLPESTKAPELDDRNAVASLKKDTAWTGVDWSNGIGVTPEEVG
                     IYIAFLNKVGFLPPPTHNDKLPLPSIELTQAQISLVASGAGARGSSAAA"
     gene            complement(717..4895)
                     /gene="LYS2"
                     /label=LYS2
     promoter        complement(4896..5135)
                     /gene="S. cerevisiae LYS2"
                     /label=LYS2 promoter
     rep_origin      complement(5189..5644)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(5427..6002)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSSELTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
                     FPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     5789..5805
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5812..5830
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5839..5946
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     5863..5879
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5913..5929)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5959..5977)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5998..6014)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6022..6038
                     /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(6046..6076)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(6400..6988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7159..8019)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8020..8124)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8151..9493
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(9114..9161)
                     /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)."