LOCUS       sequence-479467-        4864 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Tag S. pombe genes at 3' end (C-terminal end of protein) with 
            mKate2, codon-optimized for S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4864)
  TITLE     Hauf Lab Plasmids 2025
REFERENCE   2  (bases 1 to 4864)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pFA6a-pmKate2-kanMX6
FEATURES             Location/Qualifiers
     source          1..4864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      777..964
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(832..856)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1039..2395
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1039..1382
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1383..2192
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1450..1469)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2198..2395
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2499..2518)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2500..2518)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2605..2622)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2776..3364)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2856..2875)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3535..4326,4327..4395))
                     /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     4158..4177
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4396..4500)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4568..4586
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4624..4646)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4746..4765
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4846..4864
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4846..4863
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gtttctgaat tgattaagga aaacatgcat atgaagcttt acatggaagg aacagttaac
      121 aaccatcatt ttaagtgtac atctgaagga gaaggtaaac cttacgaagg tactcaaaca
      181 atgagaatta aggctgttga aggtggacct ttgccatttg cttttgatat tcttgcaact
      241 tcttttatgt acggatcaaa gacttttatt aaccatacac aaggtattcc tgatttcttt
      301 aagcaatctt ttccagaagg ttttacttgg gaaagagtta ctacatatga agatggtgga
      361 gttttgactg ctacacaaga tacatcactt caagatggtt gtcttatcta taacgttaag
      421 attcgaggag ttaactttcc ttctaacggt ccagttatgc aaaagaaaac tttaggatgg
      481 gaagcttcaa ctgaaacact ttatcctgca gatggtggat tggaaggtag agctgatatg
      541 gcattaaaac ttgttggtgg aggtcatttg atttgtaatc ttaagactac ataccgatct
      601 aaaaagcctg ctaaaaactt aaagatgcca ggagtttact acgttgatcg tagacttgaa
      661 cgtattaaag aagcagataa ggaaacttac gttgaacaac atgaagttgc tgttgcacga
      721 tactgcgatc ttccatctaa attgggtcat cgttaaggcg cgccacttct aaataagcga
      781 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
      841 aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc
      901 tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta
      961 ccggcagatc cgctagggat aacagggtaa tatagatctg tttagcttgc ctcgtccccg
     1021 ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg
     1081 cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta
     1141 taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc
     1201 tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac
     1261 gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat
     1321 atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa
     1381 ccatgggtaa ggaaaagact cacgtttcga ggccgcgatt aaattccaac atggatgctg
     1441 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     1501 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     1561 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     1621 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     1681 ccggcaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     1741 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     1801 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     1861 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     1921 tgcataagct tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     1981 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     2041 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     2101 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     2161 agtttcattt gatgctcgat gagtttttct aatcagtact gacaataaaa agattcttgt
     2221 tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa
     2281 tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag
     2341 tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc
     2401 gattcgatac taacgccgcc atccagttta aacgagctcg aattcatcga tgatatcaga
     2461 tccactagtg gcctatgcgg ccgcggatct gccggtctcc ctatagtgag tcgtattaat
     2521 ttcgataagc caggttaacc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2581 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2641 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2701 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2761 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2821 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2881 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2941 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3001 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3061 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3121 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3181 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3241 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3301 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3361 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3421 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3481 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3541 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3601 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3661 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3721 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3781 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3841 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3901 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3961 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4021 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4081 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4141 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4201 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4261 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4321 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4381 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4441 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4501 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     4561 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     4621 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4681 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     4741 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atggacatat tgtcgttaga
     4801 acgcggctac aattaataca taaccttatg tatcatacac atacgattta ggtgacacta
     4861 taga
//
