LOCUS       sequence-149443-        3911 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 3911)
  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 3911)
  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-natCX
FEATURES             Location/Qualifiers
     source          1..3911
                     /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"
     enhancer        418..721
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        722..925
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     876..896
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        970..988
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1004..1576
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1582..1779
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(1886..1904)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1991..2008)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2162..2750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2242..2261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2921..3712,3713..3781))
                     /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     3544..3563
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3782..3886)
                     /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 ctcgcgacgt
      421 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      481 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      541 ggtggactat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      601 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      661 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      721 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      781 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      841 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      901 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
      961 tatcgaaatt aatacgactc actataggga gacccaagct tacatgggta ccactcttga
     1021 cgacacggct taccggtacc gcaccagtgt cccgggggac gccgaggcca tcgaggcact
     1081 ggatgggtcc ttcaccaccg acaccgtctt ccgcgtcacc gccaccgggg acggcttcac
     1141 cctgcgggag gtgccggtgg acccgcccct gaccaaggtg ttccccgacg acgaatcgga
     1201 cgacgaatcg gacgacgggg aggacggcga cccggactcc cggacgttcg tcgcgtacgg
     1261 ggacgacggc gacctggcgg gcttcgtggt cgtctcgtac tccggctgga accgccggct
     1321 gaccgtcgag gacatcgagg tcgccccgga gcaccggggg cacggggtcg ggcgcgcgtt
     1381 gatggggctc gcgacggagt tcgcccgcga gcggggcgcc gggcacctct ggctggaggt
     1441 caccaacgtc aacgcaccgg cgatccacgc gtaccggcgg atggggttca ccctctgcgg
     1501 cctggacacc gccctgtacg acggcaccgc ctcggacggc gagcaggcgc tctacatgag
     1561 catgccctgc ccctaatcag tactgacaat aaaaagattc ttgttttcaa gaacttgtca
     1621 tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat
     1681 attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata
     1741 tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc
     1801 cgccatccag tgtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct
     1861 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
     1921 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1981 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2041 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2101 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2161 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2221 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2281 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2341 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2401 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2461 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2521 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2581 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     2641 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2701 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2761 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2821 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2881 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2941 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3001 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3061 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3121 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3181 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3241 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3301 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3361 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3421 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3481 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3541 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3601 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3661 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3721 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3781 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3841 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3901 aagtgccacc t
//
