LOCUS       sequence-79427-e        8217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  an integrating (YIp) vector with a LYS2 selectable marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8217)
  AUTHORS   Eriksson P, Thomas LR, Thorburn A, Stillman DJ
  TITLE     pRS yeast vectors with a LYS2 marker.
  JOURNAL   Biotechniques. 2004 Feb;36(2):212-3.
  PUBMED    14989082
REFERENCE   2  (bases 1 to 8217)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Biotechniques."; date: "2004-02"; volume: "36(2)"; pages: "212-3"
COMMENT     Sequence Label: pRS307
FEATURES             Location/Qualifiers
     source          1..8217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(717..4895)
                     /codon_start=1
                     /gene="S. cerevisiae LYS2"
                     /product="alpha-aminoadipate reductase, required for lysine
                     biosynthesis"
                     /label=LYS2
                     /note="yeast auxotrophic marker"
                     /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(4896..5135)
                     /gene="S. cerevisiae LYS2"
                     /label=LYS2 promoter
     rep_origin      complement(5192..5647)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5329..5350)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5541..5560
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5773..5795
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5787..5804
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5788..5804
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5811..5830
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5811..5829
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5838..5945
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     5862..5878
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5862..5878
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5912..5928)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5913..5929)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5957..5975)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5996..6012)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5996..6012)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6009..6031)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6020..6036
                     /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(join(6044..6050,6051..6068,6069..6074))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(6227..6244)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6398..6986)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7157..7948,7949..8017))
                     /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"
     primer_bind     7780..7799
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8018..8122)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8190..8208
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
      241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
      301 gacttagatt ggtatatata cgcatatgca ccatatagct tcgcaagtat tcattttaga
      361 cccatggtgg aaccctagtg ttgaatggca aagtggtgat agagttcata gaattggtca
      421 gtatcgacct gtgaagatca caaggttttg cattgaggat agtatagaag caagaatcat
      481 tgaattacag gaaaaaaagg caaatatgat tcatgctaca ataaaccaag atgaagctgc
      541 cattagcaga ctaacgccag ctgatttaca gttcttattc aataactaat attttattct
      601 cttattatat attattctcg gagtttttaa gtgacatcac ccgaaaagaa gctaagtctt
      661 tctcctaatt catatttaat tattgtacat ggacatatca tacgtaatgc tcaaccttaa
      721 gctgctgcgg agcttccacg agcaccagca cctgaagcaa ctagacttat ttgcgcttga
      781 gttagttcta tacttggcag tggaagtttg tcattatgag ttggtggagg taaaaatcca
      841 accttgttta aaaatgcaat atatatacca acctcttctg gagtaacacc tattccatta
      901 gaccaatcaa cacctgtcca tgcggtgtct ttctttaaag atgccacggc gttcctatcg
      961 tctagttccg gagctttggt actttcaggt aagttgtcta agaccatgtg tagcaaagga
     1021 tacaacgcat tttcttcatt cctgtcaata acagacgcct ccaatgattt cttccattta
     1081 gaatagcttt cgatttcgac atcgtaacca taatcgtgta aagtatacaa gtagtctttg
     1141 aataatattc ttgggtgacc cgttacttga gcaacggcca attcattttc tttgggagga
     1201 ttcaaagacg tagcaacaac aacacgagca acatgatcta ctggaaccat attcacggaa
     1261 ttttcgatat ctggaatctt acctaattgg actgaacctt tcaaaaatct caataagaaa
     1321 tcatctgtgt ttgaagaacc attggcagag gcacctgtta cgtaacctgg tctgacaata
     1381 cacccacgta ggcccctttc acctgcacgt ctaatgatgt actcagcagc ccatttggac
     1441 tgaccatatc caccagtgag cccgcttgca gagttcatta aatcgtctga ttctaaaatg
     1501 cctggcttcc cttcgctaac aagtttatct gacaaattaa agtagtattc agtgtcaaga
     1561 gtagaagtgg aggaaacaaa gtcaaagaac tttggcttgc cgacggcggc taagctcata
     1621 acattgatag ttgaaataac atttggatcc ctcaatttgg catatggata aacccagtga
     1681 actaacgcac cattatggat aattatatca actgtgtttg ccaaatccat ccacttctca
     1741 tctgaaagac caaattggct tttagataaa tcgcctaata caactttaat atttgaggca
     1801 aatttttcgt tccaagtacc ataggtgata cctgcctttt gtaatcttgc aaatgcagct
     1861 tcttcatcct tggccctgac gtgggcaaac actttgaaac tgtagttctt tggagaacgt
     1921 cctaacaaat ctgcaaggat gtaggagccc agaaatcctg tgacaccggt aacaaacaca
     1981 ttaattgttg tttttccttc ggcactatta ggttcaacaa aatattctcg agaggggtac
     2041 gaacttggta gcgtctcaac caatttcttg gcgtcttccg cataatttgc agtgacattt
     2101 tcgacgacct caccttgaga tgatccaccc gatgatttaa ttctgtcaat ttccgcggca
     2161 aaggccttta tcgttggata cttgaaaatt gtgcccaatg gtaaatcaac ttgcagcttt
     2221 ttctttaagg taaaaatcat tttggtagcc aagatagaat gaccacctaa atcgaaaaac
     2281 gaatcatctg gtgatacaga tgctggcttg gtaggtaata tacttaacca taagtctcta
     2341 acctcgcgct caacattggt aaactgagag tcgtcagttt cagaaactgt attttcagct
     2401 accaaattta attgcttggg agttgggaat tgaagtttag gcttatcaac tttaccattt
     2461 ggattcaatg gtagtttatc cataaccaca atcaaggaag gcatagcata gctagccaat
     2521 cttttcttta agaaagtcct gatgtccttg gataaaagat ggtaaccgat taagccctta
     2581 actatagggt cagtttcaac ctcctttgga acatcacttt ggaacttaga caagtcatct
     2641 ggcttgtcaa atcttgggac cataaatgtg atcaatgttg gctcattgtc ggcatttttg
     2701 cgaactaaag taatgttttc tcttaccaat ggatgttggg aaatgtgcgt atctatttct
     2761 cctaattcga ttctgaaccc acgaatttta acttgatcat cagccctacc gcaacattca
     2821 cagtcaccgt ttggtagata acgacctaaa tcacccgttc tgtacaatct atctcttgga
     2881 cctaaccaga attgtctcca aggttcacca ttatccttat ccaaataatt ccagtgatct
     2941 ttttcaacaa accagttgtt cacaaatttt tctttattca attctggtaa tcctctataa
     3001 ccttcggcca aaccacctgc acgaacataa atctcaccta tttcgccaat accacatatt
     3061 tgagtacgat cgttcctgtt aacaactagt agctgaacgt tcaacatacc tttaccagca
     3121 ggcatgacat ctttcaattt tttcaaaaag tttggatcgt cattttttga tttaacttcg
     3181 aaataagaaa ctgcacgctg tgtttcagtg gtaccgtaca tattaacaat acggcaattt
     3241 tctgccaagg tttgtaacct cagacaatca cgttttgtta aaatgtcacc cacaaagaac
     3301 gcatgatgta acttagggaa tggtgtagta gcttgggcag taagtaattg acccatggca
     3361 ggtgttaaat gggtaactgt gcaaccatac ttactcatcc attccgctaa acggcccggt
     3421 gtaccaatat catcttgagt agggacatac aattgggcac ctaaaaataa tggtgtaaac
     3481 atatctcttt gaattggatc atgtgcaata ccgctcagca ttgtgaattt atcattttct
     3541 gttaagttga accttttgga catccaattg aaataataag ccaaggaaaa atgtctacca
     3601 agaacaccct taggaatacc ttcggaacca gatgtgaaag atagggttgg gttggaatct
     3661 ggtccaacta caacacctgt tctggtgtct ttgtagtgat catatggagc caaaacatcc
     3721 tcgccattgt ccaatttgcc accttcaatg gtaccatttt cttgaatagc gatggaattg
     3781 attcttgaaa caatctccaa ttcatcattg atgtaatctt ctactagttg atccaattgt
     3841 ccagcagctc taataacaat caacccacgt ggtttagcaa cacctaagta aatggtttgt
     3901 ctggctgggg gatatgcagg gtcgataact gaaaaggttg cgccggcttt caagacaccc
     3961 atcacacata ccatcaaatc cacaccccta gaagaataga tcatcactac atcacctctt
     4021 ttgatacctg ttttaatcaa ataatgggca actatgttag aagtgcggtt gatgtcgcga
     4081 taagtgaaag aacgggactt gtcggaattt agtgttggag tctccacaac acaggttctc
     4141 tctgggaagg cttcagcatt gtcctggaaa atgtcgtgaa tacaccccac gaaatcgcac
     4201 cagcccaagt tcttagttgg atcaggtaaa ctatccttgg atgatgcggt gatcagagag
     4261 attttagtta tgcaattgga tggatcgctt agcgcagcag tcaaatattg agtaaattgg
     4321 tccgcaacaa tggttactct ttcattcgaa tacagtaagc tgttataaat taagttcaaa
     4381 acatgcgcat tattactggt atccaaattc aaagcaaagt ccactaaatg atgcttgaac
     4441 tcgtctaatt tgaaatcttg gttttccaaa aaggccaaac ggaacaactg aggggtcctt
     4501 tccagatctt ggcaactttg aattttttca gctagctcgt caaaggaaaa attggcctca
     4561 atagaattga gcttgttcaa ctcattgtta attgtagaat acagctcatt aaatgaccac
     4621 gttggttgaa tattgaatct taagattttg ttattcgcaa tataaagaac aatatcatcg
     4681 tcaccggtta ctctatatat caatgcagcc catacactca aagcgacagc gtatttgtta
     4741 gaaaaactat catgaggcac atcgagctga ggtagctgta acgaatatgt agcttgtttc
     4801 gtataaggtt cttgttgtgg gcgtaaaaag tcatgtggta acactgaaag agttggatta
     4861 tccaacttct ctatccagac cttttcgtta gtcattagtg agtaactctg tgatatctct
     4921 ctataattag cagtttttca ctgaaattcc aggaaaggta ataaactcag attttttttt
     4981 atactattgg ctgcttgtta cttatatatc ttgaacttct cccagcgggt cttcaaatac
     5041 atttgggcga tgttcatgtt cattaggcag gtatttcgac attgagtcac acgcgaaaaa
     5101 ccgccggaat tttttatgta attgcaagtg gaattgatct atgcggtgtg aaataccgca
     5161 cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa
     5221 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5281 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5341 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5401 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5461 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5521 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5581 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     5641 ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5701 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     5761 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact
     5821 cactataggg cgaattggag ctccaccgcg gtggcggccg ctctagaact agtggatccc
     5881 ccgggctgca ggaattcgat atcaagctta tcgataccgt cgacctcgag ggggggcccg
     5941 gtaccagctt ttgttccctt tagtgagggt taattccgag cttggcgtaa tcatggtcat
     6001 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata ggagccggaa
     6061 gcataaagtg taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc
     6121 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6181 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6241 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6301 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6361 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc ggcccccctg
     6421 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6481 gataccaggc gttcccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6541 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     6601 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6661 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6721 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6781 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6841 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6901 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6961 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7021 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7081 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7141 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7201 tatttcgttc atccatagtt gcctgactgc ccgtcgtgta gataactacg atacgggagg
     7261 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7321 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7381 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7441 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7501 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7561 tgttgtgaaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7621 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7681 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7741 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7801 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7861 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7921 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7981 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8041 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8101 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     8161 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
