LOCUS       sequence-311213-        4894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast codon 
            optimized tSapphire under KanMX selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4894)
  TITLE     Stelling Lab Plasmids
REFERENCE   2  (bases 1 to 4894)
  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: FRP2010
FEATURES             Location/Qualifiers
     source          1..4894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      826..1013
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(881..905)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1088..2444
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1088..1431
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1432..2241
                     /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(1499..1518)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2247..2444
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2548..2567)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2549..2567)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2654..2671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2825..3413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2905..2924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3584..4375,4376..4444))
                     /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     4207..4226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4445..4549)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4617..4635
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4673..4695)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4795..4814
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgacg
       61 gatcggtgac ggtgctggtt taattaacat gtccaagggt gaagaattat tcaccggtgt
      121 tgttccaatc ttggttgaat tggatggtga tgttaacggt cacaagtttt ctgtttctgg
      181 tgaaggtgaa ggtgatgcta cttatggtaa attgaccttg aagttcatct gtaccactgg
      241 taaattgcca gttccatggc caactttggt tactactttt tcttacggtg ttatggtttt
      301 cgccagatac ccagatcata tgaagcaaca tgatttcttc aagtccgcta tgccagaagg
      361 ttacgttcaa gaaagaacca tcttcttcaa ggatgacggt aactacaaaa ctagagccga
      421 agttaagttc gaaggtgata ctttggttaa cagaatcgaa ttgaagggta tcgacttcaa
      481 agaagatggt aacatcttgg gtcacaagtt ggagtacaac ttcaactccc ataacgttta
      541 catcatggcc gataagcaaa agaacggtat taaggccaac ttcaagatca gacacaacat
      601 tgaagatggt ggtgttcaat tggctgatca ctatcaacaa aacaccccaa ttggtgatgg
      661 tccagttttg ttgccagata accattactt gtccattcaa tccgctttgt ctaaggaccc
      721 aaacgaaaaa agagatcaca tggtcttgtt ggaattcgtt actgctgctg gtattactca
      781 tggtatggac gaattataca agtaaggcgc gccacttcta aataagcgaa tttcttatga
      841 tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt
      901 gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg
      961 ttgctttctc aggtatagta tgaggtcgct cttattgacc acacctctac cggcagatcc
     1021 gctagggata acagggtaat atagatctgt ttagcttgcc tcgtccccgc cgggtcaccc
     1081 ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg
     1141 ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc
     1201 atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac
     1261 ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct
     1321 gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt
     1381 taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac catgggtaag
     1441 gaaaagactc acgtttcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     1501 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg
     1561 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     1621 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     1681 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggcaaaaca
     1741 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     1801 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     1861 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     1921 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt
     1981 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2041 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2101 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2161 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2221 atgctcgatg agtttttcta atcagtactg acaataaaaa gattcttgtt ttcaagaact
     2281 tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga
     2341 tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag
     2401 taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact
     2461 aacgccgcca tccagtttaa acgagctcga attcatcgat gatatcagat ccactagtgg
     2521 cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc
     2581 aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2641 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2701 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2761 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2821 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2881 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2941 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3001 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3061 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3121 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3181 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3241 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3301 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3361 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3421 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3481 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3541 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3601 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3661 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3721 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3781 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3841 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3901 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3961 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4021 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4081 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4141 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4201 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4261 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4321 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4381 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4441 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4501 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4561 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     4621 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     4681 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     4741 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     4801 cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa cgcggctaca
     4861 attaatacat aaccttatgt atcatacaca tacg
//
