LOCUS       sequence-146249-        5162 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 5162)
  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 5162)
  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-TKnatAX
FEATURES             Location/Qualifiers
     source          1..5162
                     /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..2827
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      2833..3030
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3136..3155)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3137..3155)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3242..3259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3413..4001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3493..3512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4172..4963,4964..5032))
                     /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     4795..4814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5033..5137)
                     /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 tcgtggtggg aggattgggg
     1921 acagctttcg gggacggccg tgccgcccca gggtgccgag ccccagagca acgcgggccc
     1981 acgaccccat atcggggaca cgttatttac cctgtttcgg gcccccgagt tgctggcccc
     2041 caacggcgac ctgtataacg tgtttgcctg ggccttggac gtcttggcca aacgcctccg
     2101 tcccatgcac gtctttatcc tggattacga ccaatcgccc gccggctgcc gggacgccct
     2161 gctgcaactt acctccggga tggtccagac ccacgtcacc accccaggct ccataccgac
     2221 gatctgcgac ctggcgcgca cgtttgcccg ggccatgggt accactcttg acgacacggc
     2281 ttaccggtac cgcaccagtg tcccggggga cgccgaggcc atcgaggcac tggatgggtc
     2341 cttcaccacc gacaccgtct tccgcgtcac cgccaccggg gacggcttca ccctgcggga
     2401 ggtgccggtg gacccgcccc tgaccaaggt gttccccgac gacgaatcgg acgacgaatc
     2461 ggacgacggg gaggacggcg acccggactc ccggacgttc gtcgcgtacg gggacgacgg
     2521 cgacctggcg ggcttcgtgg tcgtctcgta ctccggctgg aaccgccggc tgaccgtcga
     2581 ggacatcgag gtcgccccgg agcaccgggg gcacggggtc gggcgcgcgt tgatggggct
     2641 cgcgacggag ttcgcccgcg agcggggcgc cgggcacctc tggctggagg tcaccaacgt
     2701 caacgcaccg gcgatccacg cgtaccggcg gatggggttc accctctgcg gcctggacac
     2761 cgccctgtac gacggcaccg cctcggacgg cgagcaggcg ctctacatga gcatgccctg
     2821 cccctaatca gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag
     2881 tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt
     2941 cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc
     3001 aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca
     3061 gtgtttaaac gagctcgaat tcatcgatga tatcagatcc actagtggcc tatgcggccg
     3121 cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc
     3181 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3241 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3301 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3361 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3421 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3481 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3541 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3601 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3661 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3721 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3781 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3841 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3901 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3961 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4021 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4081 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4141 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4201 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4261 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4321 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4381 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4441 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4501 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4561 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4621 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4681 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4741 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4801 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4861 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4921 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4981 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5041 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5101 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5161 ct
//
