LOCUS       40924_37848        8601 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRS317, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8601)
  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 8601)
  AUTHORS   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 8601)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8601)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "2004"; volume: "36"; issue: "2"; pages: 
            "212-213"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-SEP-2002) Pathology, University of Utah, 30 North 1900 East Rm 
            5C126 SOM, Salt Lake City, UT 84132-2501, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(571..4746)
                     /codon_start=1
                     /label=LYS2
                     /note="alpha-aminoadipate reductase, required for lysine 
                     biosynthesis"
                     /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"
     promoter        complement(4747..4986)
                     /label=LYS2 promoter
     rep_origin      complement(5040..5495)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     5640..5656
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5663..5681
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5690..5797
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(5810..5828)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5849..5865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5873..5889)
                     /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(5897..5927)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5942..5963)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6251..6839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7013..7870)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7871..7975)
                     /label=AmpR promoter
     misc_feature    8015..8518
                     /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 accatatagc ttcgcaagta ttcattttag acccatggtg gaaccctagt gttgaatggc
      241 aaagtggtga tagagttcat agaattggtc agtatcgacc tgtgaagatc acaaggtttt
      301 gcattgagga tagtatagaa gcaagaatca ttgaattaca ggaaaaaaag gcaaatatga
      361 ttcatgctac aataaaccaa gatgaagctg ccattagcag actaacgcca gctgatttac
      421 agttcttatt caataactaa tattttattc tcttattata tattattctc ggagttttta
      481 agtgacatca cccgaaaaga agctaagtct ttctcctaat tcatatttaa ttattgtaca
      541 tggacatatc atacgtaatg ctcaacctta agctgctgcg gagcttccac gagcaccagc
      601 acctgaagca actagactta tttgcgcttg agttagttct atacttggca gtggaagttt
      661 gtcattatga gttggtggag gtaaaaatcc aaccttgttt aaaaatgcaa tatatatacc
      721 aacctcttct ggagtaacac ctattccatt agaccaatca acacctgtcc atgcggtgtc
      781 tttctttaaa gatgccacgg cgttcctatc gtctagttcc ggagctttgg tactttcagg
      841 taagttgtct aagaccatgt gtagcaaagg atacaacgca ttttcttcat tcctgtcaat
      901 aacagacgcc tccaatgatt tcttccattt agaatagctt tcgatttcga catcgtaacc
      961 ataatcgtgt aaagtataca agtagtcttt gaataatatt cttgggtgac ccgttacttg
     1021 agcaacggcc aattcatttt ctttgggagg attcaaagac gtagcaacaa caacacgagc
     1081 aacatgatct actggaacca tattcacgga attttcgata tctggaatct tacctaattg
     1141 gactgaacct ttcaaaaatc tcaataagaa atcatctgtg tttgaagaac cattggcaga
     1201 ggcacctgtt acgtaacctg gtctgacaat acacccacgt aggccccttt cacctgcacg
     1261 tctaatgatg tactcagcag cccatttgga ctgaccatat ccaccagtga gcccgcttgc
     1321 agagttcatt aaatcgtctg attctaaaat gcctggcttc ccttcgctaa caagtttatc
     1381 tgacaaatta aagtagtatt cagtgtcaag agtagaagtg gaggaaacaa agtcaaagaa
     1441 ctttggcttg ccgacggcgg ctaagctcat aacattgata gttgaaataa catttggatc
     1501 cctcaatttg gcatatggat aaacccagtg aactaacgca ccattatgga taattatatc
     1561 aactgtgttt gccaaatcca tccacttctc atctgaaaga ccaaattggc ttttagataa
     1621 atcgcctaat acaactttaa tatttgaggc aaatttttcg ttccaagtac cataggtgat
     1681 acctgccttt tgtaatcttg caaatgcagc ttcttcatcc ttggccctga cgtgggcaaa
     1741 cactttgaaa ctgtagttct ttggagaacg tcctaacaaa tctgcaagga tgtaggagcc
     1801 cagaaatcct gtgacaccgg taacaaacac attaattgtt gtttttcctt cggcactatt
     1861 aggttcaaca aaatattctc gagaggggta cgaacttggt agcgtctcaa ccaatttctt
     1921 ggcgtcttcc gcataatttg cagtgacatt ttcgacgacc tcaccttgag atgatccacc
     1981 cgatgattta attctgtcaa tttccgcggc aaaggccttt atcgttggat acttgaaaat
     2041 tgtgcccaat ggtaaatcaa cttgcagctt tttctttaag gtaaaaatca ttttggtagc
     2101 caagatagaa tgaccaccta aatcgaaaaa cgaatcatct ggtgatacag atgctggctt
     2161 ggtaggtaat atacttaacc ataagtctct aacctcgcgc tcaacattgg taaactgaga
     2221 gtcgtcagtt tcagaaactg tattttcagc taccaaattt aattgcttgg gagttgggaa
     2281 ttgaagttta ggcttatcaa ctttaccatt tggattcaat ggtagtttat ccataaccac
     2341 aatcaaggaa ggcatagcat agctagccaa tcttttcttt aagaaagtcc tgatgtcctt
     2401 ggataaaaga tggtaaccga ttaagccctt aactataggg tcagtttcaa cctcctttgg
     2461 aacatcactt tggaacttag acaagtcatc tggcttgtca aatcttggga ccataaatgt
     2521 gatcaatgtt ggctcattgt cggcattttt gcgaactaaa gtaatgtttt ctcttaccaa
     2581 tggatgttgg gaaatgtgcg tatctatttc tcctaattcg attctgaacc cacgaatttt
     2641 aacttgatca tcagccctac cgcaacattc acagtcaccg tttggtagat aacgacctaa
     2701 atcacccgtt ctgtacaatc tatctcttgg acctaaccag aattgtctcc aaggttcacc
     2761 attatcctta tccaaataat tccagtgatc tttttcaaca aaccagttgt tcacaaattt
     2821 ttctttattc aattctggta atcctctata accttcggcc aaaccacctg cacgaacata
     2881 aatctcacct atttcgccaa taccacatat ttgagtacga tcgttcctgt taacaactag
     2941 tagctgaacg ttcaacatac ctttaccagc aggcatgaca tctttcaatt ttttcaaaaa
     3001 gtttggatcg tcattttttg atttaacttc gaaataagaa actgcacgct gtgtttcagt
     3061 ggtaccgtac atattaacaa tacggcaatt ttctgccaag gtttgtaacc tcagacaatc
     3121 acgttttgtt aaaatgtcac ccacaaagaa cgcatgatgt aacttaggga atggtgtagt
     3181 agcttgggca gtaagtaatt gacccatggc aggtgttaaa tgggtaactg tgcaaccata
     3241 cttactcatc cattccgcta aacggcccgg tgtaccaata tcatcttgag tagggacata
     3301 caattgggca cctaaaaata atggtgtaaa catatctctt tgaattggat catgtgcaat
     3361 accgctcagc attgtgaatt tatcattttc tgttaagttg aaccttttgg acatccaatt
     3421 gaaataataa gccaaggaaa aatgtctacc aagaacaccc ttaggaatac cttcggaacc
     3481 agatgtgaaa gatagggttg ggttggaatc tggtccaact acaacacctg ttctggtgtc
     3541 tttgtagtga tcatatggag ccaaaacatc ctcgccattg tccaatttgc caccttcaat
     3601 ggtaccattt tcttgaatag cgatggaatt gattcttgaa acaatctcca attcatcatt
     3661 gatgtaatct tctactagtt gatccaattg tccagcagct ctaataacaa tcaacccacg
     3721 tggtttagca acacctaagt aaatggtttg tctggctggg ggatatgcag ggtcgataac
     3781 tgaaaaggtt gcgccggctt tcaagacacc catcacacat accatcaaat ccacacccct
     3841 agaagaatag atcatcacta catcacctct tttgatacct gttttaatca aataatgggc
     3901 aactatgtta gaagtgcggt tgatgtcgcg ataagtgaaa gaacgggact tgtcggaatt
     3961 tagtgttgga gtctccacaa cacaggttct ctctgggaag gcttcagcat tgtcctggaa
     4021 aatgtcgtga atacacccca cgaaatcgca ccagcccaag ttcttagttg gatcaggtaa
     4081 actatccttg gatgatgcgg tgatcagaga gattttagtt atgcaattgg atggatcgct
     4141 tagcgcagca gtcaaatatt gagtaaattg gtccgcaaca atggttactc tttcattcga
     4201 atacagtaag ctgttataaa ttaagttcaa aacatgcgca ttattactgg tatccaaatt
     4261 caaagcaaag tccactaaat gatgcttgaa ctcgtctaat ttgaaatctt ggttttccaa
     4321 aaaggccaaa cggaacaact gaggggtcct ttccagatct tggcaacttt gaattttttc
     4381 agctagctcg tcaaaggaaa aattggcctc aatagaattg agcttgttca actcattgtt
     4441 aattgtagaa tacagctcat taaatgacca cgttggttga atattgaatc ttaagatttt
     4501 gttattcgca atataaagaa caatatcatc gtcaccggtt actctatata tcaatgcagc
     4561 ccatacactc aaagcgacag cgtatttgtt agaaaaacta tcatgaggca catcgagctg
     4621 aggtagctgt aacgaatatg tagcttgttt cgtataaggt tcttgttgtg ggcgtaaaaa
     4681 gtcatgtggt aacactgaaa gagttggatt atccaacttc tctatccaga ccttttcgtt
     4741 agtcattagt gagtaactct gtgatatctc tctataatta gcagtttttc actgaaattc
     4801 caggaaaggt aataaactca gatttttttt tatactattg gctgcttgtt acttatatat
     4861 cttgaacttc tcccagcggg tcttcaaata catttgggcg atgttcatgt tcattaggca
     4921 ggtatttcga cattgagtca cacgcgaaaa accgccggaa ttttttatgt aattgcaagt
     4981 ggaattatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     5041 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5101 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5161 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5221 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5281 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5341 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     5401 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     5461 acccgccgcg cttaatgcgc cgctacaggg cgcgtcgcgc cattcgccat tcaggctgcg
     5521 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaggg
     5581 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     5641 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
     5701 cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaattcg atatcaagct
     5761 tatcgatacc gtcgacctcg agggggggcc cggtacccag cttttgttcc ctttagtgag
     5821 ggttaattcc gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5881 cgctcacaat tccacacaac ataggagccg gaagcataaa gtgtaaagcc tggggtgcct
     5941 aatgagtgag gtaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6001 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6061 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6181 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6241 tgctggcgtt tttccatagg ctcggccccc ctgacgagca tcacaaaaat cgacgctcaa
     6301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgttcccc cctggaagct
     6361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6421 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     6481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6661 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6901 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6961 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7021 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7081 tgcccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7141 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7201 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7261 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7321 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7381 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg aaaaaaagcg gttagctcct
     7441 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7501 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7561 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7621 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7681 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7741 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7801 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7861 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7981 ttccccgaaa agtgccacct ggtcggtcct tttcatcacg tgctataaaa ataattataa
     8041 tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag
     8101 aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac
     8161 taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct
     8221 gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat
     8281 gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg
     8341 ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc
     8401 tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt
     8461 cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg
     8521 tcgatccgtc gacctgctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     8581 gtatcacgag gccctttcgt c
//
