LOCUS       sequence-489754-        4761 bp    DNA     circular SYN 20-OCT-2025
DEFINITION  Yeast tagging vector to create a gene fusion with yeast optimized 
            mTurquoise2 with SpHis5 selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4761)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4761)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pFA6a-link-ymTurquoise2-SpHis5
FEATURES             Location/Qualifiers
     source          1..4761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     327..346
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(565..669)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     737..755
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(793..815)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     915..934
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1015..1033
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1015..1032
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1863..2050
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1918..1942)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            2125..3325
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        2125..2468
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2469..3122
                     /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      3128..3325
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3429..3448)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3430..3448)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3535..3552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3706..4294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3786..3805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4465..4761,1..495,496..564))
                     /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"
ORIGIN
        1 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
       61 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
      121 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
      181 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
      241 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
      301 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
      361 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
      421 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
      481 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
      541 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
      601 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
      661 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
      721 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
      781 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
      841 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
      901 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt
      961 gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag
     1021 gtgacactat agaacgcggc cgccagctga agcttcgtac gctgcaggtc gacggatcgg
     1081 tgacggtgct ggtttaatta acatggttag taaaggtgaa gaattgttca ctggtgtagt
     1141 gccaatttta gtagaattag atggtgacgt taacggtcat aagttctctg taagcggtga
     1201 aggtgaaggt gacgcaacat acggtaaatt aactttgaaa ttcatttgta ctacaggtaa
     1261 attaccagtt ccatggccaa ctttggttac tactttgtct tggggtgttc aatgtttcgc
     1321 tagataccca gatcatatga agcaacatga tttctttaag tctgctatgc cagaaggtta
     1381 cgttcaagaa agaactatct tcttcaagga tgatggtaac tataagacta gagctgaagt
     1441 taagttcgaa ggagatactt tagttaatag aattgaattg aagggtattg attttaagga
     1501 agatggtaat attttgggtc ataagttgga atataattac ttctctgata acgtttacat
     1561 tactgctgat aagcaaaaga acggtattaa ggcgaacttc aagattagac ataacattga
     1621 agatggtggt gttcaattag ctgatcatta ccaacaaaac actccaattg gtgacggtcc
     1681 agttttgttg ccagataacc attacttgtc tactcaatct aagttgtcta aggatccaaa
     1741 cgaaaagaga gatcatatgg ttttgttgga attcgttact gctgctggga tcactttagg
     1801 tatggatgaa ttgtataaat aagcggccgc ctcgaggtcg acggcgcgcc acttctaaat
     1861 aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg
     1921 tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact
     1981 ctttcctgta ggtcaggttg ctttctcagg tatagtatga ggtcgctctt attgaccaca
     2041 cctctaccgg cagatccgct agggataaca gggtaatata gatctgttta gcttgcctcg
     2101 tccccgccgg gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt
     2161 gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc
     2221 catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac
     2281 ggctcctcgc tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt
     2341 ccccacgccg cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc
     2401 tttcatatac ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat
     2461 aaacaaccat gggtaggagg gcttttgtag aaagaaatac gaacgaaacg aaaatcagcg
     2521 ttgccatcgc tttggacaaa gctcccttac ctgaagagtc gaattttatt gatgaactta
     2581 taacttccaa gcatacaaac caaaagggag aacaagtaat ccaagtagac acgggaattg
     2641 gattcttgga tcacatgtat catgcactgg ctaaacatgc aggctggagc ttacgacttt
     2701 actcaagagg tgatttaatc atcgatgatc atcacactgc agaagatact gctattgcac
     2761 ttggtattgc attcaagcag gctatgagta actttgccgg cgttaaaaga tttggacatg
     2821 cttattgtcc acttgacgaa gctctttcta gaagcgtagt tgacttgtcg ggacggccct
     2881 atgctgttat cgatttggga ttaaagcgtg aaaaggttgg ggaattgtcc tgtgaaatga
     2941 tccctcactt actatattcc ttttcggtag cagctggaat tactttgcat gttacctgct
     3001 tatatggtag taatgaccat catcgtgctg aaagcgcttt taaatctctg gctgttgcca
     3061 tgcgcgcggc tactagtctt actggaagtt ctgaagtccc aagcacgaag ggagtgttgt
     3121 aaagagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt
     3181 ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct
     3241 cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg
     3301 tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagttta
     3361 aacgagctcg aattcatcga tgatatcaga tccactagtg gcctatgcgg ccgcggatct
     3421 gccggtctcc ctatagtgag tcgtattaat ttcgataagc caggttaacc tgcattaatg
     3481 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3541 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3601 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3661 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3721 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3781 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3841 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3901 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3961 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4021 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4081 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4141 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4201 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4261 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4321 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4381 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4441 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4501 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4561 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4621 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4681 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4741 ttcgccagtt aatagtttgc g
//
