LOCUS       sequence-150488-        5397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fission yeast drug counter-selectable selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5397)
  AUTHORS   Amelina H, Moiseeva V, Collopy LC, Pearson SR, Armstrong CA, Tomita 
            K
  TITLE     Sequential and counter-selectable cassettes for fission yeast.
  JOURNAL   BMC Biotechnol. 2016 Nov 8;16(1):76.
  PUBMED    27825338
REFERENCE   2  (bases 1 to 5397)
  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: "BMC 
            Biotechnol."; date: "2016-11-8"; volume: "16(1)"; pages: "76"
COMMENT     Sequence Label: pFA6a-TKanAX
FEATURES             Location/Qualifiers
     source          1..5397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..61
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(99..121)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     221..240
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        321..339
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     321..338
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2255..3064
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2322..2341)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3070..3267
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3371..3390)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3372..3390)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3477..3494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3648..4236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3728..3747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4407..5198,5199..5267))
                     /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     5030..5049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5268..5372)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
       61 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
      121 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
      181 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
      241 ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc
      301 ttatgtatca tacacatacg atttaggtga cactatagaa cgcggccgcc agctgaagct
      361 tcgtacgctg caggtcgacg gatccccggg ttaattaagg cgcgccagat ctcgaaaccc
      421 tacaacaact aatggctatc atgcggaagc tggtgagaag aacagcatcg ggacaaggga
      481 aggaagaaca aagacaaaga aaacaaaaga aagcaattga aaacaaaaca aaacaatttt
      541 cattccttct cttatcattc cttttctttt cttttctctc attcaacgca ctccatcgta
      601 tccgtattcc tcttattttt tctctttctc tatatccatt tctttctctc taggtgtgtc
      661 ctctctctct cttcaatttc tctactccgc attccaacgc atccttcccc caacctccca
      721 tttcctcctt acggcccgat agcgatcgtc tttccctcgc tatcactcgc taccggcccc
      781 tcctctgcac cgtaacctcc tacgtattta ccatatcata aagttttttc cgacgcttat
      841 cgctgacccc ctgtcgccct cctattggct tccggattat cttcttgtcc ataaggtgat
      901 ccatgcttcc tgaagattcc cgaaatgtgt ccactttggc ggggaatcat tccatccact
      961 tctttctctc tcgctttcct cattcggcgc tccccttccg cgtctcattg gtcttccgct
     1021 ccgtttttgc tttgccgatg ttacttgggg agaggtgcga taatcctttc gcaaaaactc
     1081 ggtttgacgc ctcccatggt ataaatagtg ggtggtggac aggtgccttc gcttttcttt
     1141 tctttaagca agagaagctt ccatggaaac tagtgcgttc gaccaggctg cgcgttctcg
     1201 cggccatagc aaccgacgta cggcgttgcg ccctcgccgg cagcaagaag ccacggaagt
     1261 ccgcctggag cagaaaatgc ccacgctact gcgggtttat atagacggtc ctcacgggat
     1321 ggggaaaacc accaccacgc aactgctggt ggccctgggt tcgcgcgacg atatcgtcta
     1381 cgtacccgag ccgatgactt actggcaggt gctgggggct tccgagacaa tcgcgaacat
     1441 ctacaccaca caacaccgcc tcgaccaggg tgagatatcg gccggggacg cggcggtggt
     1501 aatgacaagc gcccagataa caatgggcat gccttatgcc gtgaccgacg ccgttctggc
     1561 tcctcatatc gggggggagg ctgggagctc acatgccccg cccccggccc tcaccctcat
     1621 cttcgaccgc catcccatcg ccgccctcct gtgctacccg gccgcgcgat accttatggg
     1681 cagcatgacc ccccaggccg tgctggcgtt cgtggccctc atcccgccga ccttgcccgg
     1741 cacaaacatc gtgttggggg cccttccgga ggacagacac atcgaccgcc tggccaaacg
     1801 ccagcgcccc ggcgagcggc ttgacctggc tatgctggcc gcgattcgcc gcgtttacgg
     1861 gctgcttgcc aatacggtgc ggtatctgca gggcggcggg tcgtggcggg aggattgggg
     1921 acagctttcg gggacggccg tgccgcccca gggtgccgag ccccagagca acgcgggccc
     1981 acgaccccat atcggggaca cgttatttac cctgtttcgg gcccccgagt tgctggcccc
     2041 caacggcgac ctgtacaacg tgtttgcctg ggccttggac gtcttggcca aacgcctccg
     2101 tcccatgcac gtctttatcc tggattacga ccaatcgccc gccggctgcc gggacgccct
     2161 gctgcaactt acctccggga tggtccagac ccacgtcacc acccccggct ccataccgac
     2221 gatctgcgac ctggcgcgca cgtttgcccg ggccatgggt aaggaaaaga ctcacgtttc
     2281 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2341 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
     2401 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2461 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2521 tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc aggtattaga
     2581 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2641 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2701 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2761 tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga
     2821 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2881 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2941 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3001 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3061 ctaatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt
     3121 ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc
     3181 ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat
     3241 cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatccagtt
     3301 taaacgagct cgaattcatc gatgatatca gatccactag tggcctatgc ggccgcggat
     3361 ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa cctgcattaa
     3421 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3481 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3541 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3601 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3661 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3721 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3781 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3841 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3901 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3961 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4021 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4081 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4141 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4201 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4261 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4321 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4381 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4441 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4501 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4561 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4621 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4681 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4741 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4801 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4861 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4921 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4981 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5041 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5101 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5161 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5221 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5281 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5341 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacct
//
