LOCUS       sequence-160582-        5646 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 5646)
  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 5646)
  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-FHyTkAX
FEATURES             Location/Qualifiers
     source          1..5646
                     /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)."
     terminator      3279..3476
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    complement(3513..3546)
                     /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(3620..3639)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3621..3639)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3726..3743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3897..4485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3977..3996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4656..5447,5448..5516))
                     /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     5279..5298
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5517..5621)
                     /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 agcttacatg
     1201 tccatgggta aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     1261 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     1321 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     1381 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     1441 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     1501 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     1561 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     1621 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     1681 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     1741 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     1801 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     1861 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     1921 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     1981 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     2041 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     2101 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     2161 gatggctgtg tagaagtagc tagtgcgttc gaccaggctg cgcgttctcg cggccatagc
     2221 aaccgacgta cggcgttgcg ccctcgccgg cagcaagaag ccacggaagt ccgcctggag
     2281 cagaaaatgc ccacgctact gcgggtttat atagacggtc ctcacgggat ggggaaaacc
     2341 accaccacgc aactgctggt ggccctgggt tcgcgcgacg atatcgtcta cgtacccgag
     2401 ccgatgactt actggcaggt gctgggggct tccgagacaa tcgcgaacat ctacaccaca
     2461 caacaccgcc tcgaccaggg tgagatatcg gccggggacg cggcggtggt aatgacaagc
     2521 gcccagataa caatgggcat gccttatgcc gtgaccgacg ccgttctggc tcctcatatc
     2581 gggggggagg ctgggagctc acatgccccg cccccggccc tcaccctcat cttcgaccgc
     2641 catcccatcg ccgccctcct gtgctacccg gccgcgcgat accttatggg cagcatgacc
     2701 ccccaggccg tgctggcgtt cgtggccctc atcccgccga ccttgcccgg cacaaacatc
     2761 gtgttggggg cccttccgga ggacagacac atcgaccgcc tggccaaacg ccagcgcccc
     2821 ggcgagcggc ttgacctggc tatgctggcc gcgattcgcc gcgtttacgg gctgcttgcc
     2881 aatacggtgc ggtatctgca gggcggcggg tcgtggcggg aggattgggg acagctttcg
     2941 gggacggccg tgccgcccca gggtgccgag ccccagagca acgcgggccc acgaccccat
     3001 atcggggaca cgttatttac cctgtttcgg gcccccgagt tgctggcccc caacggcgac
     3061 ctgtataacg tgtttgcctg ggccttggac gtcttggcca aacgcctccg tcccatgcac
     3121 gtctttatcc tggattacga ccaatcgccc gccggctgcc gggacgccct gctgcaactt
     3181 acctccggga tggtccagac ccacgtcacc accccaggct ccataccgac gatctgcgac
     3241 ctggcgcgca cgtttgcccg gccctgcccc taatcagtac tgacaataaa aagattcttg
     3301 ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa
     3361 atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa
     3421 gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt
     3481 cgattcgata ctaacgccgc catcctaggc ctataacttc gtatagcata cattatacga
     3541 acggtagttt aaacgagctc gaattcatcg atgatatcag atccactagt ggcctatgcg
     3601 gccgcggatc tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac
     3661 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3721 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3781 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3841 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3901 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3961 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4021 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4081 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4141 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4201 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4261 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4321 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4381 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4441 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4501 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4561 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4621 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4681 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4741 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4801 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4861 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4921 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4981 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5041 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5101 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5161 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5221 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5281 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5341 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5401 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5461 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5521 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5581 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5641 ccacct
//
