LOCUS       Exported                4732 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast tagging vector to create a gene fusion with yeast optimized 
            mNeongreen with SpHIS5 selection.
ACCESSION   .
VERSION     .
KEYWORDS    pFA6a-link-ymNeongreen-SpHis5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4732)
  AUTHORS   Botman D, de Groot DH, Schmidt P, Goedhart J, Teusink B
  TITLE     In vivo characterisation of fluorescent proteins in budding yeast.
  JOURNAL   Sci Rep. 2019 Feb 19;9(1):2234. doi: 10.1038/s41598-019-38913-z.
  PUBMED    30783202
REFERENCE   2  (bases 1 to 4732)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4732
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     102..121
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        202..220
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     202..219
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      545..554
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     terminator      1021..1208
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1076..1100)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1283..2483
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1283..1626
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1627..2280
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHTNQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMSNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      2286..2483
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2587..2606)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2588..2606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2693..2710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2864..3452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2944..2963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3623..4483)
                     /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     4246..4265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4484..4588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4656..4674
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4712..2)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
       61 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
      121 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
      181 cttatgtatc atacacatac gatttaggtg acactataga acgcggccgc cagctgaagc
      241 ttcgtacgct gcaggtcgac ggatcggtga cggtgctggt ttaattaaca tggtctctaa
      301 gggtgaagaa gataacatgg cttctttgcc agccacccac gagttgcaca tcttcggttc
      361 cattaacggt gtcgacttcg atatggtcgg tcaaggtacc ggtaacccaa acgacggtta
      421 cgaagaattg aacttgaagt ctaccaaggg cgacttgcag ttctcccctt ggattttggt
      481 tccacacatc ggttacggtt tccaccaata cttgccatac ccagacggta tgtccccatt
      541 ccaagccgcc atggttgacg gttctggtta ccaagtccac agaaccatgc aattcgaaga
      601 cggtgcttcc ttgaccgtga actacagata cacctacgaa ggttctcaca tcaagggcga
      661 agcccaagtt aagggtaccg gtttccctgc tgacggtcca gtcatgacta actccttgac
      721 cgctgctgac tggtgtcgtt ccaagaagac ctacccaaac gataagacca tcatttccac
      781 tttcaagtgg tcctacacca ccggtaacgg caagagatac cgctctaccg ctcgtaccac
      841 ctacaccttc gccaagccaa tggctgccaa ctacttgaag aaccagccta tgtacgtctt
      901 ccgtaagacc gagctgaagc attccaagac tgaattgaac ttcaaggaat ggcaaaaggc
      961 tttcaccgac gtcatgggta tggacgaatt gtacaagtaa ggcgcgccac ttctaaataa
     1021 gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta
     1081 tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct
     1141 ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat tgaccacacc
     1201 tctaccggca gatccgctag ggataacagg gtaatataga tctgtttagc ttgcctcgtc
     1261 cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga
     1321 cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca
     1381 tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg
     1441 ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc
     1501 ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt
     1561 tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa
     1621 acaaccatgg gtaggagggc ttttgtagaa agaaatacga acgaaacgaa aatcagcgtt
     1681 gccatcgctt tggacaaagc tcccttacct gaagagtcga attttattga tgaacttata
     1741 acttccaagc atacaaacca aaagggagaa caagtaatcc aagtagacac gggaattgga
     1801 ttcttggatc acatgtatca tgcactggct aaacatgcag gctggagctt acgactttac
     1861 tcaagaggtg atttaatcat cgatgatcat cacactgcag aagatactgc tattgcactt
     1921 ggtattgcat tcaagcaggc tatgagtaac tttgccggcg ttaaaagatt tggacatgct
     1981 tattgtccac ttgacgaagc tctttctaga agcgtagttg acttgtcggg acggccctat
     2041 gctgttatcg atttgggatt aaagcgtgaa aaggttgggg aattgtcctg tgaaatgatc
     2101 cctcacttac tatattcctt ttcggtagca gctggaatta ctttgcatgt tacctgctta
     2161 tatggtagta atgaccatca tcgtgctgaa agcgctttta aatctctggc tgttgccatg
     2221 cgcgcggcta ctagtcttac tggaagttct gaagtcccaa gcacgaaggg agtgttgtaa
     2281 agagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt
     2341 atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg
     2401 acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta
     2461 tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtttaaa
     2521 cgagctcgaa ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc
     2581 cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa
     2641 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2701 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2761 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2821 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2881 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2941 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3001 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3061 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3121 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3181 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3241 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3301 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3361 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3421 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3481 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3541 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3601 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3661 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3721 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3781 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3841 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3901 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3961 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4021 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4081 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4141 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4201 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4261 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4321 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4381 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4441 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4501 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4561 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4621 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     4681 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc cc
//
