LOCUS       sequence-160578-        4289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fission yeast drug selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4289)
  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 4289)
  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-hygML
FEATURES             Location/Qualifiers
     source          1..4289
                     /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"
     promoter        452..795
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(2263..2282)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2264..2282)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2369..2386)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2540..3128)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2620..2639)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3299..4090,4091..4159))
                     /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     3922..3941
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4160..4264)
                     /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 ctgtttagct
      421 tgcctcgtcc ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga
      481 cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat
      541 acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa
      601 aaattacggc tcctcgctgc ggacctgcga gcagggaaac gctcccctca cagacgcgtt
      661 gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag
      721 gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc
      781 cgaacataaa caaccatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt
      841 ctgatcgaaa agttcgacag cgtctccgac ctgatgcggc tctcggaggg cgaagaatct
      901 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc
      961 gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt
     1021 ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt
     1081 gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg
     1141 gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc
     1201 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt
     1261 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc
     1321 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc
     1381 atgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc
     1441 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc
     1501 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg
     1561 gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc
     1621 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac
     1681 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg
     1741 gccgtctgga ccgatggctg tgtagaagta gctagctaat cgattagctg cagtaatcct
     1801 tgcttaaaaa gattctcttt ttttatgata tttgtacata aactttataa atgaaattca
     1861 taatagaaac gacacgaaat tacaaaatgg aatatgttca tagggtagac gaaactatat
     1921 acgcaatcta catacattta tcaagaagga gaaaaaggag gatgtaaagg aatacaggta
     1981 agcaaattga tactaatggc tcaacgtgat aaggaaaaag aattgcactt taacattaat
     2041 attgacaagg aggagggcac cacacaaaaa gttaggtgta acagaaaatc atgaaactat
     2101 gattcctaat ttatatattg gaggattttc tctaaaaaaa aaaaaataca acaaataaaa
     2161 aacactcaat gacctgacca tttgatggag tttaaacgag ctcgaattca tcgatgatat
     2221 cagatccact agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat
     2281 taatttcgat aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2341 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2401 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2461 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2521 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2581 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2641 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2701 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     2761 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2821 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2881 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2941 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3001 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3061 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3121 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3181 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3241 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3301 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3361 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3421 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3481 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3541 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3601 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3661 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3721 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3781 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3841 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3901 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3961 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4021 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4081 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4141 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4201 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4261 cgcgcacatt tccccgaaaa gtgccacct
//
