LOCUS       sequence-452412-        4989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create a gene fusion with yeast optimized 
            miRFP680 with SpHis5 selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4989)
  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: pFA6a-link-ymiRFP680-SpHis5
FEATURES             Location/Qualifiers
     source          1..4989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     329..348
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(567..671)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     739..757
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(795..817)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     917..936
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1017..1035
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1017..1034
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      2093..2280
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2148..2172)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            2355..3555
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        2355..2698
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2699..3352
                     /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      3358..3555
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3659..3678)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3660..3678)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3765..3782)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3936..4524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4016..4035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4695..4989,1..497,498..566))
                     /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 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
       61 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
      121 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
      181 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
      241 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
      301 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
      361 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
      421 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
      481 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
      541 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
      601 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
      661 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
      721 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
      781 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
      841 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
      901 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata
      961 ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt
     1021 aggtgacact atagaacgcg gccgccagct gaagcttcgt acgctgcagg tcgacggatc
     1081 ggtgacggtg ctggtttaat taacatggct gaaggttctg ttgctagaca accagatttg
     1141 ttgacttgtg atgatgaacc aattcatatt ccaggtgcta ttcaaccaca tggtttgtta
     1201 ttagctttgg ctgctgatat gaccattgtt gctggttctg ataacttgcc tgaattgact
     1261 ggtttagcta ttggtgcctt gatcggtaga tctgctgctg atgttttcga ttctgaaacc
     1321 cataatagat tgactattgc tttggctgaa ccaggtgctg ctgtcggtgc tccaattacc
     1381 gtcggtttca ccatgcgtaa ggatgccggt ttcattggtt cttggcatag acatgatcaa
     1441 ttaattttct tggaattgga accaccacaa cgtgacgttg ccgaaccaca ggctttcttc
     1501 cgtagaacca actctgctat tagaagatta caagctgctg aaactttgga atctgcttgt
     1561 gctgctgctg ctcaagaagt tcgtaagatt actggtttcg atagagttat gatctacaga
     1621 tttgcttctg atttctctgg tgaagttatt gctgaagata gatgtgctga agttgaatct
     1681 aagttgggtt tgcattaccc agcttctact gttccagctc aagctagaag attgtacact
     1741 attaatccag ttagaattat tccagatatc aactacagac cagtcccagt taccccagat
     1801 ttgaacccag ttactggtag acctattgat ttatctttcg ctattttgag atctgtttcc
     1861 ccatgtcatt tggaattcat gagaaacatc ggtatgcatg gtaccatgtc tatttctatt
     1921 ttgagaggtg aaagattgtg gggtttgatt gtttgtcatc atagaactcc atactacgtt
     1981 gacttggatg gtagacaagc ctgtaagaga gttgctgaaa gattggctac ccaaattggt
     2041 gttatggaag aataagcggc cgcctcgagg tcggcgcgcc acttctaaat aagcgaattt
     2101 cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt
     2161 ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta
     2221 ggtcaggttg ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg
     2281 cagatccgct agggataaca gggtaatata gatctgttta gcttgcctcg tccccgccgg
     2341 gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca
     2401 gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat
     2461 catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc
     2521 tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg
     2581 cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac
     2641 ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat
     2701 gggtaggagg gcttttgtag aaagaaatac gaacgaaacg aaaatcagcg ttgccatcgc
     2761 tttggacaaa gctcccttac ctgaagagtc gaattttatt gatgaactta taacttccaa
     2821 gcatacaaac caaaagggag aacaagtaat ccaagtagac acgggaattg gattcttgga
     2881 tcacatgtat catgcactgg ctaaacatgc aggctggagc ttacgacttt actcaagagg
     2941 tgatttaatc atcgatgatc atcacactgc agaagatact gctattgcac ttggtattgc
     3001 attcaagcag gctatgagta actttgccgg cgttaaaaga tttggacatg cttattgtcc
     3061 acttgacgaa gctctttcta gaagcgtagt tgacttgtcg ggacggccct atgctgttat
     3121 cgatttggga ttaaagcgtg aaaaggttgg ggaattgtcc tgtgaaatga tccctcactt
     3181 actatattcc ttttcggtag cagctggaat tactttgcat gttacctgct tatatggtag
     3241 taatgaccat catcgtgctg aaagcgcttt taaatctctg gctgttgcca tgcgcgcggc
     3301 tactagtctt actggaagtt ctgaagtccc aagcacgaag ggagtgttgt aaagagtact
     3361 gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta
     3421 gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc
     3481 tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg
     3541 ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagttta aacgagctcg
     3601 aattcatcga tgatatcaga tccactagtg gcctatgcgg ccgcggatct gccggtctcc
     3661 ctatagtgag tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa
     3721 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3781 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3841 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3901 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3961 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4021 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4081 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4141 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4201 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4261 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4321 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4381 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4441 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4501 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4561 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4621 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4681 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4741 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4801 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4861 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4921 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4981 aatagtttg
//
