LOCUS       sequence-146247-        4981 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 4981)
  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 4981)
  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-TKnatCX
FEATURES             Location/Qualifiers
     source          1..4981
                     /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        447..750
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        751..954
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     905..925
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     951..975
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1072..1119
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2227..2799
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      2805..3002
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3061..3078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3232..3820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3312..3331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3991..4782,4783..4851))
                     /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     4614..4633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4852..4956)
                     /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 ctgagctcgc
      421 ggccgcgata tcgctagctc gaggtccgtt acataactta cggtaaatgg cccgcctggc
      481 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      541 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      601 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      661 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      721 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      781 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      841 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      901 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      961 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     1021 cgggaccgat ccagcctccg cggccgggaa cggtgcattg gaacggaccg tgttgacaat
     1081 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     1141 gctagtgcgt tcgaccaggc tgcgcgttct cgcggccata gcaaccgacg tacggcgttg
     1201 cgccctcgcc ggcagcaaga agccacggaa gtccgcctgg agcagaaaat gcccacgcta
     1261 ctgcgggttt atatagacgg tcctcacggg atggggaaaa ccaccaccac gcaactgctg
     1321 gtggccctgg gttcgcgcga cgatatcgtc tacgtacccg agccgatgac ttactggcag
     1381 gtgctggggg cttccgagac aatcgcgaac atctacacca cacaacaccg cctcgaccag
     1441 ggtgagatat cggccgggga cgcggcggtg gtaatgacaa gcgcccagat aacaatgggc
     1501 atgccttatg ccgtgaccga cgccgttctg gctcctcata tcggggggga ggctgggagc
     1561 tcacatgccc cgcccccggc cctcaccctc atcttcgacc gccatcccat cgccgccctc
     1621 ctgtgctacc cggccgcgcg ataccttatg ggcagcatga ccccccaggc cgtgctggcg
     1681 ttcgtggccc tcatcccgcc gaccttgccc ggcacaaaca tcgtgttggg ggcccttccg
     1741 gaggacagac acatcgaccg cctggccaaa cgccagcgcc ccggcgagcg gcttgacctg
     1801 gctatgctgg ccgcgattcg ccgcgtttac gggctgcttg ccaatacggt gcggtatctg
     1861 cagggcggcg ggtcgtggtg ggaggattgg ggacagcttt cggggacggc cgtgccgccc
     1921 cagggtgccg agccccagag caacgcgggc ccacgacccc atatcgggga cacgttattt
     1981 accctgtttc gggcccccga gttgctggcc cccaacggcg acctgtataa cgtgtttgcc
     2041 tgggccttgg acgtcttggc caaacgcctc cgtcccatgc acgtctttat cctggattac
     2101 gaccaatcgc ccgccggctg ccgggacgcc ctgctgcaac ttacctccgg gatggtccag
     2161 acccacgtca ccaccccagg ctccataccg acgatctgcg acctggcgcg cacgtttgcc
     2221 cgggccatgg gtaccactct tgacgacacg gcttaccggt accgcaccag tgtcccgggg
     2281 gacgccgagg ccatcgaggc actggatggg tccttcacca ccgacaccgt cttccgcgtc
     2341 accgccaccg gggacggctt caccctgcgg gaggtgccgg tggacccgcc cctgaccaag
     2401 gtgttccccg acgacgaatc ggacgacgaa tcggacgacg gggaggacgg cgacccggac
     2461 tcccggacgt tcgtcgcgta cggggacgac ggcgacctgg cgggcttcgt ggtcgtctcg
     2521 tactccggct ggaaccgccg gctgaccgtc gaggacatcg aggtcgcccc ggagcaccgg
     2581 gggcacgggg tcgggcgcgc gttgatgggg ctcgcgacgg agttcgcccg cgagcggggc
     2641 gccgggcacc tctggctgga ggtcaccaac gtcaacgcac cggcgatcca cgcgtaccgg
     2701 cggatggggt tcaccctctg cggcctggac accgccctgt acgacggcac cgcctcggac
     2761 ggcgagcagg cgctctacat gagcatgccc tgcccctaat cagtactgac aataaaaaga
     2821 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     2881 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     2941 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     3001 tgctgtcgat tcgatactaa cgccgccatc cagtttaaac gagctcgaat tctaggcttc
     3061 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3121 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3181 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3241 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3301 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3361 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3421 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3481 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3541 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3601 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3661 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3721 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3781 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3841 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3901 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3961 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4021 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4081 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4141 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4201 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4261 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4321 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4381 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4441 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4501 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4561 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4621 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4681 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4741 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4801 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4861 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4921 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4981 t
//
