LOCUS       Exported                5554 bp ds-DNA     circular SYN 23-NOV-2020
DEFINITION  pFA6a-link-yEGFP-Kan.
ACCESSION   P30175
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5554)
  AUTHORS   Self
  JOURNAL   Unpublished.
REFERENCE   2  (bases 1 to 5554)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 23, 2020 from SnapGene 5.2.3
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished."
COMMENT     SECNOTES|Mark A. Sheff and Kurt S. Thorn.  Optimized cassettes for 
            fluorescent protein tagging in  Saccharomyces cerevisiae.  Yeast 21 
            (2004) 661-670
FEATURES             Location/Qualifiers
     source          1..5554
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..671
                     /locus_tag="TEF Promoter"
                     /label=TEF Promoter
     primer_bind     complement(649..677)
                     /label=hifi_rev_1
     misc_feature    672..677
                     /locus_tag="XhoI"
                     /label=XhoI
     misc_feature    678..680
                     /locus_tag="Start"
                     /label=Start
     misc_binding    681..688
                     /locus_tag="PacI"
                     /label=PacI
     CDS             690..1406
                     /codon_start=1
                     /locus_tag="eGFP"
                     /label=eGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     primer_bind     699..723
                     /label=hifi_for_2
     terminator      1414..1616
                     /locus_tag="ADH1 terminator"
                     /label=ADH1 terminator
     promoter        1654..2032
                     /locus_tag="Ashbya gossypii TEF promoter"
                     /label=Ashbya gossypii TEF promoter
     CDS             2033..2842
                     /gene="Kan"
                     /locus_tag="kanamycin phosphotransferase"
                     /product="kanamycin phosphotransferase"
                     /label=kanamycin phosphotransferase
     misc_feature    2851..3075
                     /gene="TEFt"
                     /locus_tag="Ashbya gossypii TEF terminator"
                     /product="Ashbya gossypii TEF terminator"
                     /label=Ashbya gossypii TEF terminator
     primer_bind     3076..3102
                     /locus_tag="DUB_ORF_INT-R"
                     /label=DUB_ORF_INT-R
     primer_bind     complement(3076..3102)
                     /label=BAC690_TEF_DUBr
     rep_origin      3410..3495
                     /locus_tag="Origin of replication"
                     /label=Origin of replication
     CDS             complement(4185..5045)
                     /gene="AmpR"
                     /locus_tag="bla, beta-lactamase"
                     /product="bla, beta-lactamase"
                     /label=bla, beta-lactamase
     primer_bind     5534..5554
                     /locus_tag="DUB_ORF_int-F"
                     /label=DUB_ORF_int-F
     primer_bind     5534..5554
                     /label=BAC690_TEF_DUBf
ORIGIN
        1 actttgttat gtagagtttt tttagctacc tatattccac cataacatca atcatgcggt
       61 tgctggtgta tttaccaata atgtttaatg tatatatata tatatatata tggggccgta
      121 tacttacata tagtagatgt caagcgtagg cgcttcccct gccggctgtg agggcgccat
      181 aaccaaggta tctatagacc gccaatcagc aaactacctc cgtacattca tgttgcaccc
      241 acacatttat acacccagac cgcgacaaat tacccataag gttgtttgtg acggcgtcgt
      301 acaagagaac gtgggaactt tttaggctca ccaaaaaaga aagaaaaaat acgagttgct
      361 gacagaagcc tcaagaaaaa aaaaattctt cttcgactat gctggaggca gagatgatcg
      421 agccggtagt taactatata tagctaaatt ggttccatca ccttcttttc tggtgtcgct
      481 ccttctagtg ctatttctgg cttttcctat tttttttttt ccatttttct ttctctcttt
      541 ctaatatata aattctcttg cattttctat ttttctctct atctattcta cttgtttatt
      601 cccttcaagg ttttttttta aggagtactt gtttttagaa tatacggtca acgaactata
      661 attaactaaa cctcgagatg ttaattaaca tgtctaaagg tgaagaatta ttcactggtg
      721 ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt tctgtctccg
      781 gtgaaggtga aggtgatgct acttacggta aattgacctt aaaatttatt tgtactactg
      841 gtaaattgcc agttccatgg ccaaccttag tcactacttt aacttatggt gttcaatgtt
      901 tttctagata cccagatcat atgaaacaac atgacttttt caagtctgcc atgccagaag
      961 gttatgttca agaaagaact atttttttca aagatgacgg taactacaag accagagctg
     1021 aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt attgatttta
     1081 aagaagatgg taacatttta ggtcacaaat tggaatacaa ctataactct cacaatgttt
     1141 acatcatggc tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt agacacaaca
     1201 ttgaagatgg ttctgttcaa ttagctgacc attatcaaca aaatactcca attggtgatg
     1261 gtccagtctt gttaccagac aaccattact tatccactca atctgcctta tccaaagatc
     1321 caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct ggtattaccc
     1381 atggtattga tgaattgtac aaataaggcg cgccacttct aaataagcga atttcttatg
     1441 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
     1501 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
     1561 gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta ccggcagatc
     1621 cgctagggat aacagggtaa tatagatctg tttagcttgc ctcgtccccg ccgggtcacc
     1681 cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag
     1741 gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg
     1801 catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga
     1861 cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc
     1921 tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt
     1981 ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtaa
     2041 ggaaaagact cacgtttcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2101 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg
     2161 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2221 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2281 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggcaaaac
     2341 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2401 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2461 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     2521 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataagct
     2581 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     2641 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2701 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2761 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2821 gatgctcgat gagtttttct aatcagtact gacaataaaa agattcttgt tttcaagaac
     2881 ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg
     2941 atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa
     3001 gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac
     3061 taacgccgcc atccagttta aacgagctcg aattcatcga tgatatcaga tccactagtg
     3121 gcctatgcgg ccgcggatct gccggtctcc ctatagtgag tcgtattaat ttcgataagc
     3181 caggttaacc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3241 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3301 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3361 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3421 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3481 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3541 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3601 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3661 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3721 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3781 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3841 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     3901 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3961 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4021 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4081 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4141 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4201 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4261 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4321 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4381 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4441 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4501 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4561 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4621 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4681 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4741 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4801 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4861 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4921 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4981 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5041 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5101 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5161 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     5221 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     5281 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     5341 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     5401 tcagagcaga ttgtactgag agtgcaccat atggacatat tgtcgttaga acgcggctac
     5461 aattaataca taaccttatg tatcatacac atacgattta ggtgacacta tagaacgcgg
     5521 ccgccagctg aagcttcgta cgctgcaggt cgac
//
