LOCUS       sequence-149574-        3673 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 3673)
  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 3673)
  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-zeoCV
FEATURES             Location/Qualifiers
     source          1..3673
                     /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"
     polyA_signal    413..494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     438..457
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(508..882)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVCVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(901..948)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(1045..1069)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1066..1269)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1095..1115)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1270..1573
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(1647..1666)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1648..1666)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1753..1770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1924..2512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2004..2023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2683..3474,3475..3543))
                     /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     3306..3325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3544..3648)
                     /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 ctgcagtgaa
      421 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
      481 gcaataaaca agtttcgagg tcgagtgtca gtcctgctcc tcggccacga agtgcacgca
      541 gttgccggcc gggtcgcgca gggcgaactc ccgcccccac ggctgctcgc cgatctcggt
      601 catggccggc ccggaggcgt cccggaagtt cgtggacacg acctccgacc actcggcgta
      661 cagctcgtcc aggccgcgca cacacaccca ggccagggtg ttgtccggca ccacctggtc
      721 ctggaccgcg ctgatgaaca gggtcacgtc gtcccggacc acaccggcga agtcgtcctc
      781 cacgaagtcc cgggagaacc cgagccggtc ggtccagaac tcgaccgctc cggcgacgtc
      841 gcgcgcggtg agcaccggaa cggcactggt caacttggcc atggtttagt tcctcacctt
      901 gtcgtattat actatgccga tatactatgc cgatgattaa ttgtcaacac ggtccgttcc
      961 aatgcaccgt tcccggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa
     1021 acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata
     1081 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     1141 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     1201 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     1261 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     1321 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     1381 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     1441 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     1501 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     1561 aagttatgta acggtttaaa cgagctcgaa ttcatcgatg atatcagatc cactagtggc
     1621 ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt cgataagcca
     1681 ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1741 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1801 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1861 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1921 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1981 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2041 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2101 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2161 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2221 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2281 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2341 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2401 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2461 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2521 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2581 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2641 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2701 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2761 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2821 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2881 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2941 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3001 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3061 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3121 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3181 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3241 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3301 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3361 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3421 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3481 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3541 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3601 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3661 aaaagtgcca cct
//
