LOCUS       Exported                4075 bp DNA     circular SYN 03-NOV-2025
DEFINITION  Plasmid for yeast gene deletion using the natMX6 selectable marker 
            conferring nourseothricin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    pFA6a-5FLAG-natMX6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4075)
  AUTHORS   Hentges P, Van Driessche B, Tafforeau L, Vandenhaute J, Carr AM.
  TITLE     Three novel antibiotic marker cassettes for gene disruption and 
            marker switching in Schizosaccharomyces pombe.
  JOURNAL   Yeast 2005;22:1013-9.
  PUBMED    16200533
REFERENCE   2  (bases 1 to 4075)
  AUTHORS   EUROSCARF
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4075)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4075)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4075)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 4075)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; 
            date: "2005"; volume: "22"; pages: "1013-9"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     SGRef: number: 5; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             67..198
                     /codon_start=1
                     /label=5xFLAG
                     /note="five tandem FLAG(R) epitope tags"
                     /translation="DYKDDDDKDYKDDDDKLMDYKDDDDKDYKDDDDKLMDYKDDDDK"
     terminator      225..412
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     gene            487..1606
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        487..830
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             831..1403
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1409..1606
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(1711..1729)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1987..2575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2746..3606)
                     /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"
     promoter        complement(3607..3711)
                     /label=AmpR promoter
     promoter        4057..4075
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaat
       61 catatggact acaaggacga cgatgacaag gattacaaag atgacgacga taagcttatg
      121 gactacaagg acgacgatga caaggattac aaagatgacg acgataagct tatggactac
      181 aaggacgacg atgacaagca tatgggcgcg ccacttctaa ataagcgaat ttcttatgat
      241 ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg
      301 actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt
      361 tgctttctca ggtatagtat gaggtcgctc ttattgacca cacctctacc ggcagatccg
      421 ctagggataa cagggtaata tagatctgtt tagcttgcct cgtccccgcc gggtcacccg
      481 gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg
      541 gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca
      601 tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc
      661 tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg
      721 tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt
      781 taaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc atgggtacca
      841 ctcttgacga cacggcttac cggtaccgca ccagtgtccc gggggacgcc gaggccatcg
      901 aggcactgga tgggtccttc accaccgaca ccgtcttccg cgtcaccgcc accggggacg
      961 gcttcaccct gcgggaggtg ccggtggacc cgcccctgac caaggtgttc cccgacgacg
     1021 aatcggacga cgaatcggac gacggggagg acggcgaccc ggactctcgg acgttcgtcg
     1081 cgtacgggga cgacggcgac ctggcgggct tcgtggtcgt ctcgtactcc ggctggaacc
     1141 gccggctgac cgtcgaggac atcgaggtcg ccccggagca ccgggggcac ggggtcgggc
     1201 gcgcgttgat ggggctcgcg acggagttcg cccgcgagcg gggtgccggg cacctctggc
     1261 tggaggtcac caacgtcaac gcaccggcga tccacgcgta ccggcggatg gggttcaccc
     1321 tctgcggcct ggacaccgcc ctgtacgacg gcaccgcctc ggacggcgag caggcgctct
     1381 acatgagcat gccctgcccc taatcagtac tgacaataaa aagattcttg ttttcaagaa
     1441 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
     1501 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
     1561 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata
     1621 ctaacgccgc catccagttt aaacgagctc gaattcatcg atgatatcag atccactagt
     1681 ggcctatgcg gccgcggatc tgccggtctc cctatagtga gtcgtattaa tttcgataag
     1741 ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1801 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1861 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1921 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1981 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2041 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2101 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2161 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2221 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2281 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2341 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2401 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2461 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2521 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2581 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2641 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2701 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2761 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2821 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2881 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2941 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3001 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3061 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3121 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3181 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3241 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3301 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3361 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3421 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3481 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3541 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3601 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3661 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3721 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3781 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     3841 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     3901 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     3961 atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag aacgcggcta
     4021 caattaatac ataaccttat gtatcataca catacgattt aggtgacact ataga
//
