LOCUS       sequence-311199-        4888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast codon 
            optimized mKate2 under KanMX selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4888)
  TITLE     Stelling Lab Plasmids
REFERENCE   2  (bases 1 to 4888)
  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"
COMMENT     Sequence Label: FRP266
FEATURES             Location/Qualifiers
     source          1..4888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            1..1357
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1..344
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             345..1154
                     /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(412..431)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1160..1357
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1461..1480)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1462..1480)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1567..1584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1738..2326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1818..1837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2497..3288,3289..3357))
                     /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     3120..3139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3358..3462)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3530..3548
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3586..3608)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3708..3727
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3808..3826
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3808..3825
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      4627..4814
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4682..4706)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
       61 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
      121 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag
      181 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
      241 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
      301 ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt aaggaaaaga
      361 ctcacgtttc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
      421 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
      481 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
      541 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
      601 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc
      661 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
      721 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
      781 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
      841 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat
      901 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
      961 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     1021 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     1081 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     1141 atgagttttt ctaatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt
     1201 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat
     1261 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc
     1321 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg
     1381 ccatccagtt taaacgagct cgaattcatc gatgatatca gatccactag tggcctatgc
     1441 ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa
     1501 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1561 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1621 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1681 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1741 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1801 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1861 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1921 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1981 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2041 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2101 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2161 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2221 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2281 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2341 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2401 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2461 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2521 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2581 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2641 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2701 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2761 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2821 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2881 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2941 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3001 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3061 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3121 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3181 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3241 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3301 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3361 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3421 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3481 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3541 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3601 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3661 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     3721 gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata
     3781 cataacctta tgtatcatac acatacgatt taggtgacac tatagaacgc ggccgccagc
     3841 tgaagcttcg tacgctgcag gtcgacggat cggtgacggt gctggtttaa ttaacaccgg
     3901 tatggtttct gaattgatta aagaaaacat gcatatgaag ttgtatatgg aaggtactgt
     3961 taacaatcat catttcaagt gtacatctga aggtgaaggt aaaccatatg aaggtactca
     4021 aactatgaga attaaagctg ttgaaggtgg tccattgcca tttgcttttg atattttggc
     4081 tacttctttc atgtatggtt ctaagacttt cattaaccat actcaaggta ttccagattt
     4141 tttcaagcaa tcttttccag aaggttttac ttgggaaaga gttactactt acgaagatgg
     4201 tggtgttttg actgctactc aagatacttc tttgcaagat ggttgtttga tttacaacgt
     4261 taagattaga ggtgttaatt ttccatctaa tggtccagtt atgcaaaaaa agactttggg
     4321 ttgggaagca tctactgaaa ctttgtatcc agctgatggt ggtttggaag gtagagctga
     4381 tatggctttg aaattggttg gtggtggtca tttgatttgt aacttgaaaa ctacttacag
     4441 atcaaaaaag ccagctaaga atttgaaaat gccaggtgtt tattacgttg atagaagatt
     4501 ggaaagaatt aaagaagctg ataaagaaac ttacgttgaa caacatgaag ttgctgttgc
     4561 aagatattgt gatttgccat ctaaattggg tcatagagcc ggctagggcg cgccacttct
     4621 aaataagcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata
     4681 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
     4741 aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac
     4801 cacacctcta ccggcagatc cgctagggat aacagggtaa tatagatctg tttagcttgc
     4861 ctcgtccccg ccgggtcacc cggccagc
//
