LOCUS       sequence-83369-e        3944 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 3944)
  AUTHORS   Goldstein AL, McCusker JH
  TITLE     Three new dominant drug resistance cassettes for gene disruption in 
            Saccharomyces cerevisiae.
  JOURNAL   Yeast. 1999 Oct;15(14):1541-53.
  PUBMED    10514571
REFERENCE   2  (bases 1 to 3944)
  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: "1999-10"; volume: "15(14)"; pages: "1541-53"
COMMENT     Sequence Label: pAG60
FEATURES             Location/Qualifiers
     source          1..3944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        115..458
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1277..1474
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1581..1600)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1582..1600)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1687..1704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1858..2446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1938..1957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2617..3408,3409..3477))
                     /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"
     primer_bind     3240..3259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3478..3582)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3650..3668
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3706..3728)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3828..3847
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(3928..3944,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(3928..3944,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
      121 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
      181 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
      241 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
      301 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
      361 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
      421 ggatacagtt ctcacatcac atccgaacat aaacaaccat gacagtcaac actaagacct
      481 atagtgagag agcagaaact catgcctcac cagtagcaca gcgattattt cgattaatgg
      541 aactgaagaa aaccaattta tgtgcatcaa ttgacgttga taccactaag gaattccttg
      601 aattaattga taaattaggt ccttatgtat gcttaatcaa gactcatatt gatataatca
      661 atgatttttc ctatgaatcc actattgaac cattattaga actttcacgt aaacatcaat
      721 ttatgatttt tgaagataga aaatttgctg atattggtaa taccgtaaag aaacaatata
      781 ttggtggagt ttataaaatt agtagttggg cagatattac caatgctcat ggtgtcactg
      841 ggaatggagt ggttgaagga ttaaaacagg gagctaaaga aaccaccacc aaccaagagc
      901 caagagggtt attgatgtta gctgaattat catcagtggg atcattagca tatggagaat
      961 attctcaaaa aactgttgaa attgctaaat ccgataagga atttgttatt ggatttattg
     1021 cccaacgtga tatgggtggc caagaagaag gatttgattg gcttattatg acacctggag
     1081 ttggattaga tgataaaggt gatggattag gacaacaata tagaactgtt gatgaagttg
     1141 ttagcactgg aactgatatt atcattgttg gtagaggatt gtttggtaaa ggaagagatc
     1201 cagatattga aggtaaaagg tatagaaatg ctggttggaa tgcttatttg aaaaagactg
     1261 gccaattata atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt
     1321 atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt
     1381 ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg
     1441 cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca
     1501 tccagtgtcg aaaacgagct cgaattcatc gatgatatca gatccactag tggcctatgc
     1561 ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa
     1621 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1681 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1741 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1801 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1861 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1921 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1981 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2041 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2101 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2161 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2221 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2281 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2341 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2401 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2461 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2521 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2581 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2641 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2701 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2761 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2821 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2881 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2941 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3001 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3061 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3121 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3181 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3241 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3301 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3361 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3421 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3481 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3541 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3601 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3661 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3721 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3781 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     3841 gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata
     3901 cataacctta tgtatcatac acatacgatt taggtgacac tata
//
