LOCUS       sequence-83389-e        7531 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7531)
  AUTHORS   Ito-Harashima S, McCusker JH
  TITLE     Positive and negative selection LYS5MX gene replacement cassettes 
            for use in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2004 Jan 15;21(1):53-61.
  PUBMED    14745782
REFERENCE   2  (bases 1 to 7531)
  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: "Yeast."; 
            date: "2004-01-15"; volume: "21(1)"; pages: "53-61"
COMMENT     Sequence Label: pSA52
FEATURES             Location/Qualifiers
     source          1..7531
                     /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"
     promoter        187..467
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             468..1142
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1239..1694)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1376..1397)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1588..1607
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1824..1846
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1838..1855
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1839..1855
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1862..1881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1862..1880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2472..2815
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3811..4008
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     4661..4677
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4661..4677
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4711..4727)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4712..4728)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4757..4775)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4796..4812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4796..4812)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4809..4831)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4820..4836
                     /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(4844..4850,4851..4868,4869..4874))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5027..5044)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5198..5786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5957..6748,6749..6817))
                     /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     6580..6599
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6818..6922)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6959..7462
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7504..7522
                     /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 accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat
      241 atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
      301 aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
      361 gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
      421 tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
      481 atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
      541 cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
      601 agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
      661 atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
      721 ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
      781 ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
      841 actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
      901 attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
      961 gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     1021 atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     1081 agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     1141 aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgcctg cggtgtgaaa
     1201 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaaacg ttaatatttt
     1261 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     1321 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     1381 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     1441 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     1501 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     1561 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     1621 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     1681 gctacagggc gcgtcgcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     1741 ggtgcgggcc tcttcgctat tacgccagct ggcgaagggg ggatgtgctg caaggcgatt
     1801 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     1861 gtaatacgac tcactatagg gcgaattgga gctccaccgc ggtggcggcc gccagctgaa
     1921 gcttcgtacg ctgcaggtcg acggatcccc gggttaatta aggcgcgcca gatctgcccg
     1981 ccgggaaggc gaacccgatc ggatgcatcc tctctgctgc catgatgctg aagttgtcgt
     2041 tgaacatggt tgctgccggc gaggcggtcg agcaggcagt gcaggaggtg ttggactcgg
     2101 gagtcagaac gggcgacctg ctcggctcga gctccacttc ggaggttggc gacgccattg
     2161 cgcttgcagt taaggaagcc ttgcgcaggc aatccgcagc tggtctgagc tagcctcgag
     2221 gacccttctc tttagactat tctactctta tgcacgtaaa aaattctagg aaatatgtat
     2281 taactaggag taaaataacc ggctagtggc attcatatag ccgtctgttt acatctacat
     2341 cacacatttc gagtgtatat ctcgcaacgt tggcgttaaa taggcagtca atggcccgac
     2401 cattctatgg tgtttaggtc gatgccatct ttgtacagct tgcctcgtcc ccgccgggtc
     2461 acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct
     2521 caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat
     2581 ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc
     2641 agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc
     2701 ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc
     2761 cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa caaccatgac
     2821 attacaagat catcttaatg ataaatcaaa tatcatccta cttttcacca ctagattaac
     2881 taatgaatta attgaatttt ggaatgatga ttataatttt gaatgttctt taagattatt
     2941 aaatgattta tcattacaac aaaaaatatc ttcaattaca aaaaacccca accatcaaca
     3001 acaaagattt aaatataaac aattattatc aaatttattt attagatatg caatcaattc
     3061 aatcattcaa tcaaatgatc tatttaaaaa aattgaattt gattataatg aatttgggaa
     3121 acccatatta ccatataatt tccaattcaa tattagtagt tctaatgata taattgcctt
     3181 aattgtggaa tttccatctt catcttcatt atcatcatca tcattgtcat catcattatc
     3241 tggtgatcga gcttatgacc caattggtat tgatttatca cattctattc aaaatagtat
     3301 atcaaataca gaatatcttg aacaatttaa accgattttt gatgatgttt atgaattacc
     3361 acaaattgat aattatttta aatttaatca tttttggaca ttaaaagaat catttacaaa
     3421 attgattggt agtggattaa atattgaatt atcagatttt tatttcatag ttgattataa
     3481 tgataatgag ttttctagtg ataatgaaaa attccaaaca agtatagttg gaaataatat
     3541 tggagatagt tggatgtatt attcattgaa ttggtttgat aaaatcttga taaatataga
     3601 taaacttgaa ttagccaaga atcaatttat aacggcatta aataaaaaag aattttattg
     3661 tcgtagttct attcttgata atggtaatgt cgatgaatct aaattgccag taattataac
     3721 gataattaat caaaacaaga ataaatcaat tcaaccaata gagttgagat ttgaaaaatt
     3781 attatctgaa catttaaacc agtgatcagt actgacaata aaaagattct tgttttcaag
     3841 aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc
     3901 gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag
     3961 aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga
     4021 tactaacgcc gccatccagt gtcgactagg gttgctgcca tcggcctcgc tcgcgtcttt
     4081 gccggatagc aagagcgcct ttggcctcta cgagccctgc cacggctctg cgcccgatct
     4141 gcccgccggg aaggcgaacc cgatcggatg catcctctct gctgccatga tgctgaagtt
     4201 gtcgttgaac atggttgctg ccggcgaggc ggtcgagcag gcagtgcagg aggtgttgga
     4261 ctcgggagtc agaacgggcg acctgctcgg ctcgagctcc acttcggagg ttggcgacgc
     4321 cattgcgctt gcagttaagg aagccttgcg caggcaatcc gcagctggtc tgagctagcc
     4381 tcgaggaccc ttctctttag actattctac tcttatgcac gtaaaaaatt ctaggaaata
     4441 tgtattaact aggagtaaaa taaccggcta gtggcattca tatagccgtc tgtttacatc
     4501 tacatcacac atttcgagtg tatatctcgc aacgttggcg ttaaataggc agtcaatggc
     4561 ccgaccattc tatggtgttt aggtcgatgc catctttgta cgtttaaacg agctcgaatt
     4621 catcgatgat atcagatcca ctagtggcct atgcggccgc tctagaacta gtggatcccc
     4681 cgggctgcag gaattcgata tcaagcttat cgataccgtc gacctcgagg gggggcccgg
     4741 tacccagctt ttgttccctt tagtgagggt taattccgag cttggcgtaa tcatggtcat
     4801 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata ggagccggaa
     4861 gcataaagtg taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc
     4921 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4981 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5041 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5101 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5161 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc ggcccccctg
     5221 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5281 gataccaggc gttcccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5341 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     5401 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5461 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5521 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5581 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     5641 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5701 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5761 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5821 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5881 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5941 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6001 tatttcgttc atccatagtt gcctgactgc ccgtcgtgta gataactacg atacgggagg
     6061 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6121 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6181 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6241 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6301 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6361 tgttgtgaaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6421 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6481 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6541 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6601 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6661 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6721 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6781 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6841 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6901 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg tccttttcat
     6961 cacgtgctat aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat
     7021 agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac
     7081 tctaggggga tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta
     7141 atgccgaatt gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta
     7201 cattttactt atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa
     7261 tatatatgta aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct
     7321 tcattccgta actcttctac cttctttatt tactttctaa aatccaaata caaaacataa
     7381 aaataaataa acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg
     7441 tgtaagttac aggcaagcga tccgtcctaa gaaaccatta ttatcatgac attaacctat
     7501 aaaaataggc gtatcacgag gccctttcgt c
//
