LOCUS       sequence-83376-e        4703 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 4703)
  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 4703)
  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: pAG31
FEATURES             Location/Qualifiers
     source          1..4703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            572..1673
                     /product="yeast selectable marker conferring bialaphos 
                     resistance (Goldstein and McCusker, 1999)"
                     /label=patMX4
     promoter        572..915
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             970..1422
                     /codon_start=1
                     /gene="Streptomyces viridochromogenes pat1"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialaphos or phosphinothricin"
                     /translation="MAAVCDIVNHYIETSTVNFRTEPQTPQEWIDDLERLQDRYPWLVA
                     EVEGVVAGIAYAGPWKARNAYDWTVESTVYVSHRHQRLGLGSTLYTHLLKSMEAQGFKS
                     VVAVIGLPNDPSVRLHEALGYTARGTLRAAGYKHGGWHDVGFWQRDF"
     terminator      1476..1673
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2340..2359)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2341..2359)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2446..2463)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2617..3205)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2697..2716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3376..4167,4168..4236))
                     /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     3999..4018
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4237..4341)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4409..4427
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4465..4487)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4587..4606
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4687..4703,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4687..4703,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatctgcccg ccgggaaggc gaacccgatc ggatgcatcc tctctgctgc
      121 catgatgctg aagttgtcgt tgaacatggt tgctgccggc gaggcggtcg agcaggcagt
      181 gcaggaggtg ttggactcgg gagtcagaac gggcgacctg ctcggctcga gctccacttc
      241 ggaggttggc gacgccattg cgcttgcagt taaggaagcc ttgcgcaggc aatccgcagc
      301 tggtctgagc tagcctcgag gacccttctc tttagactat tctactctta tgcacgtaaa
      361 aaattctagg aaatatgtat taactaggag taaaataacc ggctagtggc attcatatag
      421 ccgtctgttt acatctacat cacacatttc gagtgtatat ctcgcaacgt tggcgttaaa
      481 taggcagtca atggcccgac cattctatgg tgtttaggtc gatgccatct ttgtacagct
      541 tgcctcgtcc ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga
      601 cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat
      661 acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa
      721 aaattacggc tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt
      781 gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag
      841 gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc
      901 cgaacataaa caaccatggg tagcccagaa cgacgcccgg tcgagatccg tcccgccacc
      961 gccgccgaca tggcggcggt ctgcgacatc gtcaatcact acatcgagac gagcacggtc
     1021 aacttccgta cggagccgca gactccgcag gagtggatcg acgacctgga gcgcctccag
     1081 gaccgctacc cctggctcgt cgccgaggtg gagggcgtcg tcgccggcat cgcctacgcc
     1141 ggcccctgga aggcccgcaa cgcctacgac tggaccgtcg agtcgacggt gtacgtctcc
     1201 caccggcacc agcggctcgg actgggctcc accctctaca cccacctgct gaagtccatg
     1261 gaggcccagg gcttcaagag cgtggtcgcc gtcatcggac tgcccaacga cccgagcgtg
     1321 cgcctgcacg aggcgctcgg atacaccgcg cgcgggacgc tgcgggcagc cggctacaag
     1381 cacgggggct ggcacgacgt ggggttctgg cagcgcgact tcgagctgcc ggccccgccc
     1441 cgccccgtcc ggcccgtcac acagatctaa tcagtactga caataaaaag attcttgttt
     1501 tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg
     1561 ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg
     1621 cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga
     1681 ttcgatacta acgccgccat ccagtgtcga ctagggttgc tgccatcggc ctcgctcgcg
     1741 tctttgccgg atagcaagag cgcctttggc ctctacgagc cctgccacgg ctctgcgccc
     1801 gatctgcccg ccgggaaggc gaacccgatc ggatgcatcc tctctgctgc catgatgctg
     1861 aagttgtcgt tgaacatggt tgctgccggc gaggcggtcg agcaggcagt gcaggaggtg
     1921 ttggactcgg gagtcagaac gggcgacctg ctcggctcga gctccacttc ggaggttggc
     1981 gacgccattg cgcttgcagt taaggaagcc ttgcgcaggc aatccgcagc tggtctgagc
     2041 tagcctcgag gacccttctc tttagactat tctactctta tgcacgtaaa aaattctagg
     2101 aaatatgtat taactaggag taaaataacc ggctagtggc attcatatag ccgtctgttt
     2161 acatctacat cacacatttc gagtgtatat ctcgcaacgt tggcgttaaa taggcagtca
     2221 atggcccgac cattctatgg tgtttaggtc gatgccatct ttgtacgttt aaacgagctc
     2281 gaattcatcg atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc
     2341 cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca
     2401 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2461 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2521 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2581 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2641 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2701 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2761 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     2821 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2881 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2941 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3001 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3061 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3121 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3181 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3241 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3301 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3361 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3421 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3481 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3541 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3601 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3661 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3721 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3781 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3841 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3901 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3961 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4021 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4081 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4141 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4201 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4261 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4321 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4381 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     4441 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4501 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4561 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4621 tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca
     4681 catacgattt aggtgacact ata
//
