LOCUS       sequence-149479-        5472 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 5472)
  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 5472)
  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-FTKanAX
FEATURES             Location/Qualifiers
     source          1..5472
                     /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"
     protein_bind    complement(403..436)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2290..3099
                     /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(2357..2376)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3105..3302
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    complement(3339..3372)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3446..3465)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3447..3465)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3552..3569)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3723..4311)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3803..3822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4482..5273,5274..5342))
                     /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     5105..5124
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5343..5447)
                     /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 ttaattaaga tctaccgttc gtatagcata
      421 cattatacga agttatccta gtcatatgcc tacaacaact aagaaaatgg ctatcatgcg
      481 gaagctggtg agaagaacag catcgggaca agggaaggaa gaacaaagac aaagaaaaca
      541 aaagaaagca attgaaaaca aaacaaaaca attttcattc cttctcttat cattcctttt
      601 cttttctttt ctctcattca acgcactcca tcgtatccgt attcctctta ttttttctct
      661 ttctctatat ccatttcttt ctctctaggt gtgtcctctc tctctcttca atttctctac
      721 tccgcattcc aacgcatcct tcccccaacc tcccatttcc tccttacggc ccgatagcga
      781 tcgtctttcc ctcgctatca ctcgctaccg gcccctcctc tgcaccgtaa cctcctacgt
      841 atttaccata tcataaagtt ttttccgacg cttatcgctg accccctgtc gccctcctat
      901 tggcttccgg attatcttct tgtccataag gtgatccatg cttcctgaag attcccgaaa
      961 tgtgtccact ttggcgggga atcattccat ccacttcttt ctctctcgct ttcctcattc
     1021 ggcgctcccc ttccgcgtct cattggtctt ccgctccgtt tttgctttgc cgatgttact
     1081 tggggagagg tgcgataatc ctttcgcaaa aactcggttt gacgcctccc atggtataaa
     1141 tagtgggtgg tggacaggtg ccttcgcttt tcttttcttt aagcaagaga agcttccatg
     1201 gaaactagtg cgttcgacca ggctgcgcgt tctcgcggcc atagcaaccg acgtacggcg
     1261 ttgcgccctc gccggcagca agaagccacg gaagtccgcc tggagcagaa aatgcccacg
     1321 ctactgcggg tttatataga cggtcctcac gggatgggga aaaccaccac cacgcaactg
     1381 ctggtggccc tgggttcgcg cgacgatatc gtctacgtac ccgagccgat gacttactgg
     1441 caggtgctgg gggcttccga gacaatcgcg aacatctaca ccacacaaca ccgcctcgac
     1501 cagggtgaga tatcggccgg ggacgcggcg gtggtaatga caagcgccca gataacaatg
     1561 ggcatgcctt atgccgtgac cgacgccgtt ctggctcctc atatcggggg ggaggctggg
     1621 agctcacatg ccccgccccc ggccctcacc ctcatcttcg accgccatcc catcgccgcc
     1681 ctcctgtgct acccggccgc gcgatacctt atgggcagca tgacccccca ggccgtgctg
     1741 gcgttcgtgg ccctcatccc gccgaccttg cccggcacaa acatcgtgtt gggggccctt
     1801 ccggaggaca gacacatcga ccgcctggcc aaacgccagc gccccggcga gcggcttgac
     1861 ctggctatgc tggccgcgat tcgccgcgtt tacgggctgc ttgccaatac ggtgcggtat
     1921 ctgcagggcg gcgggtcgtg gcgggaggat tggggacagc tttcggggac ggccgtgccg
     1981 ccccagggtg ccgagcccca gagcaacgcg ggcccacgac cccatatcgg ggacacgtta
     2041 tttaccctgt ttcgggcccc cgagttgctg gcccccaacg gcgacctgta caacgtgttt
     2101 gcctgggcct tggacgtctt ggccaaacgc ctccgtccca tgcacgtctt tatcctggat
     2161 tacgaccaat cgcccgccgg ctgccgggac gccctgctgc aacttacctc cgggatggtc
     2221 cagacccacg tcaccacccc cggctccata ccgacgatct gcgacctggc gcgcacgttt
     2281 gcccgggcca tgggtaagga aaagactcac gtttcgaggc cgcgattaaa ttccaacatg
     2341 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     2401 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     2461 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     2521 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     2581 gcgatccccg gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     2641 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     2701 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     2761 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2821 aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2881 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2941 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     3001 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     3061 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagtactgac aataaaaaga
     3121 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     3181 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     3241 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     3301 tgctgtcgat tcgatactaa cgccgccatc ctaggcctat aacttcgtat agcatacatt
     3361 atacgaacgg tagtttaaac gagctcgaat tcatcgatga tatcagatcc actagtggcc
     3421 tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag
     3481 gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3541 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3601 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3661 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3721 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3781 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3841 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3901 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3961 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4021 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4081 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4141 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     4201 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4261 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4321 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4381 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4441 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4501 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4561 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4621 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4681 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4741 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4801 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4861 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4921 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4981 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5041 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5101 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5161 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5221 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5281 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5341 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5401 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5461 aaagtgccac ct
//
