LOCUS       Exported                4884 bp ds-DNA     circular SYN 15-MAY-2024
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            TagRFp657 with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4884)
  AUTHORS   Lee S, Lim WA, Thorn KS
  TITLE     Improved Blue, Green, and Red Fluorescent Protein Tagging Vectors 
            for S. cerevisiae.
  JOURNAL   PLoS One. 2013 Jul 2;8(7):e67902. doi: 10.1371/journal.pone.0067902.
            Print 2013.
  PUBMED    23844123
REFERENCE   2  (bases 1 to 4884)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4884
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     125..143
                     /locus_tag="pBRforEco"
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     primer_bind     complement(181..203)
                     /locus_tag="pGEX 3'"
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     303..322
                     /locus_tag="pRS-marker"
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     promoter        403..421
                     /locus_tag="SP6 promoter"
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     403..420
                     /locus_tag="SP6"
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    491..1177
                     /locus_tag="ymScarletI3"
                     /label=ymScarletI3
     terminator      1218..1405
                     /gene="S. cerevisiae ADH1"
                     /locus_tag="ADH1 terminator"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1273..1297)
                     /locus_tag="pHybLex-R"
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1480..2836
                     /locus_tag="kanMX"
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     promoter        1480..1823
                     /locus_tag="TEF promoter"
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1824..2633
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /locus_tag="KanR"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1891..1910)
                     /locus_tag="Kan-R"
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2639..2836
                     /locus_tag="TEF terminator"
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2940..2959)
                     /locus_tag="T7"
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2941..2959)
                     /locus_tag="T7 promoter"
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3046..3063)
                     /locus_tag="L4440"
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3217..3805)
                     /direction=LEFT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3297..3316)
                     /locus_tag="pBR322ori-F"
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3976..4836)
                     /codon_start=1
                     /gene="bla"
                     /locus_tag="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4599..4618
                     /locus_tag="Amp-R"
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4837..4884,1..57))
                     /gene="bla"
                     /locus_tag="AmpR promoter"
                     /label=AmpR promoter
ORIGIN
        1 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
       61 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
      121 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
      181 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
      241 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
      301 tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg
      361 cggctacaat taatacataa ccttatgtat catacacata cgatttaggt gacactatag
      421 aacgcggccg ccagctgaag cttcgtacgc tgcaggtcga cggatcggtg acggtgctgg
      481 tttaattaac atggattcta ctgaagctgt tattaaggaa ttcatgagat ttaaggttca
      541 tatggaaggt tctatgaatg gtcatgaatt tgaaattgaa ggtgaaggtg aaggtagacc
      601 atatgaaggt actcaaactg ctaagttgaa ggtcactaag ggtggtccat tgccattctc
      661 ttgggatatt ttgtctccac aattcatgta cggttctaga gcttttatta agcatccagc
      721 tgacattcca gattactgga agcaatcttt cccagaaggt tttaaatggg aaagagttat
      781 gatctttgaa gatggtggta ctgtttctgt tactcaagat acttctttgg aagatggtac
      841 tttgatttac aaggttaagt tgagaggtgg taacttccca ccagatggtc cagttatgca
      901 aaagagaact atgggttggg aagcctctac tgaaagattg tacccagaag atgttgtttt
      961 aaagggtgat attaagatgg ctttgagatt gaaggatggt ggtagatact tggctgattt
     1021 taagactact tataaggcta agaagccagt tcaaatgcca ggtgctttca acattgatag
     1081 aaagttggat attacttctc ataacgaaga ttacactgtt gttgaacaat acgaaagatc
     1141 tgttgctaga cattctactg gtggttctgg tggttcttaa gcggccgcct cgaggtcggc
     1201 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
     1261 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
     1321 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
     1381 ctcttattga ccacacctct accggcagat ccgctaggga taacagggta atatagatct
     1441 gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc
     1501 ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt
     1561 tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg
     1621 cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca
     1681 gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg
     1741 ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac
     1801 atcacatccg aacataaaca accatgggta aggaaaagac tcacgtttcg aggccgcgat
     1861 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     1921 aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga
     1981 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     2041 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     2101 ggttactcac cactgcgatc cccggcaaaa cagcattcca ggtattagaa gaatatcctg
     2161 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     2221 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     2281 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     2341 ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca
     2401 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     2461 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     2521 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     2581 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagtac
     2641 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     2701 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     2761 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
     2821 gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagttt aaacgagctc
     2881 gaattcatcg atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc
     2941 cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca
     3001 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3061 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3121 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3181 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3241 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3301 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3361 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     3421 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3481 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3541 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3601 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3661 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3721 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3781 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3841 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3901 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3961 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4021 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4081 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4141 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4201 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4261 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4321 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4381 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4441 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4501 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4561 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4621 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4681 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4741 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4801 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4861 aagcatttat cagggttatt gtct
//
