LOCUS       sequence-149482-        5235 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 5235)
  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 5235)
  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-FTKnatAX
FEATURES             Location/Qualifiers
     source          1..5235
                     /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..2862
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      2868..3065
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    complement(3102..3135)
                     /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(3209..3228)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3210..3228)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3315..3332)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3486..4074)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3566..3585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4245..5036,5037..5105))
                     /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     4868..4887
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5106..5210)
                     /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 gtgggaggat tggggacagc tttcggggac ggccgtgccg
     1981 ccccagggtg ccgagcccca gagcaacgcg ggcccacgac cccatatcgg ggacacgtta
     2041 tttaccctgt ttcgggcccc cgagttgctg gcccccaacg gcgacctgta taacgtgttt
     2101 gcctgggcct tggacgtctt ggccaaacgc ctccgtccca tgcacgtctt tatcctggat
     2161 tacgaccaat cgcccgccgg ctgccgggac gccctgctgc aacttacctc cgggatggtc
     2221 cagacccacg tcaccacccc aggctccata ccgacgatct gcgacctggc gcgcacgttt
     2281 gcccgggcca tgggtaccac tcttgacgac acggcttacc ggtaccgcac cagtgtcccg
     2341 ggggacgccg aggccatcga ggcactggat gggtccttca ccaccgacac cgtcttccgc
     2401 gtcaccgcca ccggggacgg cttcaccctg cgggaggtgc cggtggaccc gcccctgacc
     2461 aaggtgttcc ccgacgacga atcggacgac gaatcggacg acggggagga cggcgacccg
     2521 gactcccgga cgttcgtcgc gtacggggac gacggcgacc tggcgggctt cgtggtcgtc
     2581 tcgtactccg gctggaaccg ccggctgacc gtcgaggaca tcgaggtcgc cccggagcac
     2641 cgggggcacg gggtcgggcg cgcgttgatg gggctcgcga cggagttcgc ccgcgagcgg
     2701 ggcgccgggc acctctggct ggaggtcacc aacgtcaacg caccggcgat ccacgcgtac
     2761 cggcggatgg ggttcaccct ctgcggcctg gacaccgccc tgtacgacgg caccgcctcg
     2821 gacggcgagc aggcgctcta catgagcatg ccctgcccct aatcagtact gacaataaaa
     2881 agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat
     2941 tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg
     3001 cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta
     3061 tactgctgtc gattcgatac taacgccgcc atcctaggcc tataacttcg tatagcatac
     3121 attatacgaa cggtagttta aacgagctcg aattcatcga tgatatcaga tccactagtg
     3181 gcctatgcgg ccgcggatct gccggtctcc ctatagtgag tcgtattaat ttcgataagc
     3241 caggttaacc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3301 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3361 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3421 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3481 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3541 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3601 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3661 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3721 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3781 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3841 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3901 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     3961 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4021 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4081 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4141 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4201 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4261 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4321 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4381 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4441 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4501 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4561 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4621 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4681 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4741 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4801 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4861 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4921 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4981 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5041 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5101 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5161 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5221 cgaaaagtgc cacct
//
