LOCUS       sequence-83373-e        3704 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 3704)
  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 3704)
  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: pAG25
FEATURES             Location/Qualifiers
     source          1..3704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            115..1234
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        115..458
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             459..1031
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1037..1234
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1341..1360)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1342..1360)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1447..1464)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1618..2206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1698..1717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2377..3168,3169..3237))
                     /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     3000..3019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3238..3342)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3410..3428
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3466..3488)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3588..3607
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(3688..3704,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(3688..3704,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatctgttta gcttgccttg 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 gggtaccact cttgacgaca
      481 cggcttaccg gtaccgcacc agtgtcccgg gggacgccga ggccatcgag gcactggatg
      541 ggtccttcac caccgacacc gtcttccgcg tcaccgccac cggggacggc ttcaccctgc
      601 gggaggtgcc ggtggacccg cccctgacca aggtgttccc cgacgacgaa tcggacgacg
      661 aatcggacga cggggaggac ggcgacccgg actcccggac gttcgtcgcg tacggggacg
      721 acggcgacct ggcgggcttc gtggtcgtct cgtactccgg ctggaaccgc cggctgaccg
      781 tcgaggacat cgaggtcgcc ccggagcacc gggggcacgg ggtcgggcgc gcgttgatgg
      841 ggctcgcgac ggagttcgcc cgcgagcggg gcgccgggca cctctggctg gaggtcacca
      901 acgtcaacgc accggcgatc cacgcgtacc ggcggatggg gttcaccctc tgcggcctgg
      961 acaccgccct gtacgacggc accgcctcgg acggcgagca ggcgctctac atgagcatgc
     1021 cctgccccta atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt
     1081 atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt
     1141 ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg
     1201 cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca
     1261 tccagtgtcg aaaacgagct cgaattcatc gatgatatca gatccactag tggcctatgc
     1321 ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa
     1381 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1441 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1501 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1561 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1621 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1681 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1741 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1801 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1861 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1921 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1981 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2041 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2101 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2161 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2221 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2281 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2341 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2401 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2461 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2521 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2581 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2641 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2701 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2761 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2821 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2881 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2941 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3001 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3061 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3121 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3181 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3241 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3301 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3361 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3421 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3481 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3541 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     3601 gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata
     3661 cataacctta tgtatcatac acatacgatt taggtgacac tata
//
