LOCUS       sequence-39185-e        4545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gene deletion in S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4545)
  AUTHORS   Gadaleta MC, Iwasaki O, Noguchi C, Noma K, Noguchi E
  TITLE     New vectors for epitope tagging and gene disruption in 
            Schizosaccharomyces pombe.
  JOURNAL   Biotechniques. 2013 Nov;55(5):257-63. doi: 10.2144/000114100.
  PUBMED    24215641
REFERENCE   2  (bases 1 to 4545)
  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"; doi: "10"; journalName: 
            "Biotechniques"; date: "2013-11"; volume: "55"; issue: "5"; pages: 
            "257-63"
COMMENT     Sequence Label: pFA6a-his3MX6
FEATURES             Location/Qualifiers
     source          1..4545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(335..402,458..469,655..1266,1326..1788)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his3"
                     /product="imidazole acetol phosphate transaminase, required
                     for histidine biosynthesis"
                     /label=S. pombe his3
                     /translation="MFDLNTCLRKNILELQPYRCARDDFSEGVLLDANECAYGSVISVD
                     GVEFNRYPDPRQIEVKQRLCDLRNKELSITKPLTPDNICMGVGSDEIIDSLIRISCIPG
                     KDKILMCPPSYGMYTVSAKINDVEVVKVLLEPDFNLNVDAICETLSKDSAIKVFFACSP
                     GNPTAKALKLEDIKKILEHPTWNGIVVVDEAYIDFSAPDMSALTLVNEYPNLAVCQTLS
                     KSFGLAGIRIGFCLTSKPIATIMNSLKAPYNISEPTSRLALDALSPQSIDKMHTYRDAI
                     IQQRVRLCKELTTIKGMGKIIGGYDANFILIQVLDRPEGGKPSNDAAKYLYLQMATMHK
                     VVVRFRGTEPLCEGALRITVGTEEENTILLKTIKLVLQEYYTKK"
     primer_bind     complement(2182..2201)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2183..2201)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2288..2305)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2459..3047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2539..2558)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3218..4009,4010..4078))
                     /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     3841..3860
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4079..4183)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4251..4269
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4307..4329)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4429..4448
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4529..4545,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4529..4545,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatctctttc aacgttttct ttactattgc acttataatt gtatttggaa
      121 atgaaagaca tatggagctt tatttctctt tcctcttcag gtttctgaat ttattggata
      181 taatgacacg gtgatacacg ttgtaatgaa ttaaagactc acattgctac acactatata
      241 tttatagata tcacctgcat cttaggttaa ttattaatta atctcatcga tcgctttctt
      301 cttactttaa caaaaacaca tcgaaagaga tacgatgttt gatttgaata cttgtcttcg
      361 taaaaatatc ttagaacttc agccttatcg ctgtgcgaga gagtatgtaa taacttgata
      421 gactagaaag cgtgatatat attaacatag aatatagtga cttctctgag tatgttttat
      481 ctttatattt gggtaataat tgatatgagg gctacttcta gaaagatatt ttattagctg
      541 cttggaaagc aatgaagtac tccctgttta ctgttgttgt tggagctggc gatttcaggc
      601 ctttgttgtt ttttaccctt ttctcgccta tatatttttt gctaatctta atagaggtgt
      661 ccttcttgat gccaatgaat gtgcttatgg ttcggttatt tccgtggacg gtgtggaatt
      721 caatcgttat ccggatccta gacagatcga agtaaagcaa agattatgtg atttgagaaa
      781 caaagagctt tccattacca agccactaac accagataac atttgcatgg gtgttggcag
      841 cgatgaaatt attgactcct taattcgcat ttcttgtatt cctggaaaag ataagatttt
      901 gatgtgtcca ccttcgtacg gaatgtatac ggtctctgca aaaatcaacg atgttgaggt
      961 tgttaaggtt ctccttgaac cagatttcaa tttgaacgtc gatgctattt gcgaaacact
     1021 ttctaaagat agtgccatca aagtcttttt tgcttgttcc cctggtaatc caacagctaa
     1081 ggctcttaaa ctcgaggata ttaagaagat tttggagcac cctacatgga atggaatcgt
     1141 tgtcgtcgat gaggcctata ttgatttctc tgctcccgat atgtctgcct taactcttgt
     1201 caatgaatat ccaaaccttg ctgtttgcca aaccctttcg aaatctttcg gtctagcagg
     1261 aattcggtaa gttaatctaa gcttttaagc attaatatta tttctttact aactatgatg
     1321 tttagtatcg gattttgttt gacaagtaaa cctattgcta ccattatgaa ctcattaaag
     1381 gctccttata acattagcga acccacctct cgtttagctt tggatgcttt atcccctcag
     1441 tcgattgata aaatgcatac atatagagat gctattatac agcaaagagt tcgtctttgc
     1501 aaagagctta ctaccattaa aggaatgggc aagattatag gtggttatga tgcaaatttt
     1561 atcctcattc aggttttaga tagaccagaa ggaggtaagc ctagtaacga tgccgcaaaa
     1621 tatctttatt tacaaatggc cacgatgcac aaagtcgttg ttagatttag aggtacagag
     1681 cctctttgcg aaggagcatt gcgtatcact gttggcacag aagaagagaa tacgatatta
     1741 ctaaaaacta ttaaacttgt attgcaggaa tactatacaa aaaaatgagg tttccttttc
     1801 tggcctatta gaacgtttca attagacaaa ttataactct ttcctttccc cttccggact
     1861 cgtcacattt cttttctgat gtacccaata tattttgtgt tcgctaattg cgcttgcatt
     1921 ccttctttct ttattaaaat acgttttctt cctttttggt gtgtttccgt atagttcgat
     1981 aagcaatgaa aagatgttat ttttaatttt gtaaaggagt tacgctaatt taaaggatta
     2041 taacccgtgt aataaaagaa ttaaagattc gttagctttg catagaagat ctctaatcga
     2101 attccccggt ttaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg
     2161 cggccgcgga tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta
     2221 acctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2281 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2341 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2401 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2461 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2521 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2581 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2641 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2701 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2761 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2821 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2881 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2941 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3001 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3061 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3121 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3181 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3241 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3301 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3361 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3421 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3481 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3541 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3601 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3661 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3721 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3781 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3841 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3901 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3961 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4021 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4081 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4141 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4201 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4261 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4321 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4381 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4441 agattgtact gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat
     4501 acataacctt atgtatcata cacatacgat ttaggtgaca ctata
//
