LOCUS       sequence-97834-e        4738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast C-terminal tagging vector, which creates gene fusions with 
            GFP-gamma using SpHis5 selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4738)
  AUTHORS   Slubowski CJ, Funk AD, Roesner JM, Paulissen SM, Huang LS
  TITLE     Plasmids for C-terminal tagging in Saccharomyces cerevisiae that 
            contain improved GFP proteins, Envy and Ivy.
  JOURNAL   Yeast. 2015 Jan 22. doi: 10.1002/yea.3065.
  PUBMED    25612242
REFERENCE   2  (bases 1 to 4738)
  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"; journalName: "Yeast. 2015
            Jan 22. doi: 10.1002/yea.3065."
COMMENT     Sequence Label: pFA6a-link-GFP-gamma-SpHis5
FEATURES             Location/Qualifiers
     source          1..4738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(9..31)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     131..150
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        231..249
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     231..248
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1056..1243
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1111..1135)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1318..2518
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1318..1661
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1662..2315
                     /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="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMSNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      2321..2518
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2622..2641)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2623..2641)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2728..2745)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2899..3487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2979..2998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3658..4449,4450..4518))
                     /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     4281..4300
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4519..4623)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4691..4709
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
       61 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
      121 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgga catattgtcg
      181 ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga
      241 cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgacg gatcggtgac
      301 ggtgctggtt taattaacat gagcaagggg gaagagttgt tcactggtgt tgtgcctata
      361 ttggtcgaat tggatggaga tgttaatggt cataagtttt cagtctccgg cgagggcgaa
      421 ggagatgcga cttacggtaa gctgactctc aagtttatct gtaccacagg aaagttacca
      481 gtaccatggc caactttggt gactacgtta tgttatggcg tacaatgctt tgctagatac
      541 ccagaccaca tgaaacaaca tgatttcttc aaatctgcaa tgcctgaagg ttatgtgcag
      601 gagcgtacaa tctttttcaa agatgacggt aattacaaaa caagagctga agttaagttc
      661 gaaggtgata cccttgttaa tagaatcgaa ttgaaaggta tcgattttaa ggaggacggt
      721 aatattctag ggcacaaatt ggaatacaac tacaattcac acaacgtcta catcatggct
      781 gacaaacaga aaaatggaat taaagccaac tttaaaacaa ggcataacat tgaagatggt
      841 tcagtccaat tagccgatca ttatcaacaa aatactccaa ttggggatgg ccctgttctg
      901 ctacctgaca atcattacct ttctacacaa tctgctttaa gtaaagaccc aaacgaaaag
      961 agagatcaca tggttctatt agagtttgta acagcagcag gaataacaca tggcatggat
     1021 gaactttaca aataaggcgc gccacttcta aataagcgaa tttcttatga tttatgattt
     1081 ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg
     1141 ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc
     1201 aggtatagta tgaggtcgct cttattgacc acacctctac cggcagatcc gctagggata
     1261 acagggtaat atagatctgt ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac
     1321 atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt
     1381 gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat
     1441 tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag
     1501 ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat
     1561 ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg
     1621 ctaggataca gttctcacat cacatccgaa cataaacaac catgggtagg agggcttttg
     1681 tagaaagaaa tacgaacgaa acgaaaatca gcgttgccat cgctttggac aaagctccct
     1741 tacctgaaga gtcgaatttt attgatgaac ttataacttc caagcatgca aaccaaaagg
     1801 gagaacaagt aatccaagta gacacgggaa ttggattctt ggatcacatg tatcatgcac
     1861 tggctaaaca tgcaggctgg agcttacgac tttactcaag aggtgattta atcatcgatg
     1921 atcatcacac tgcagaagat actgctattg cacttggtat tgcattcaag caggctatga
     1981 gtaactttgc cggcgttaaa agatttggac atgcttattg tccacttgac gaagctcttt
     2041 ctagaagcgt agttgacttg tcgggacggc cctatgctgt tatcgatttg ggattaaagc
     2101 gtgaaaaggt tggggaattg tcctgtgaaa tgatccctca cttactatat tccttttcgg
     2161 tagcagctgg aattactttg catgttacct gcttatatgg tagtaatgac catcatcgtg
     2221 ctgaaagcgc ttttaaatct ctggctgttg ccatgcgcgc ggctactagt cttactggaa
     2281 gttctgaagt cccaagcacg aagggagtgt tgtaaagagt actgacaata aaaagattct
     2341 tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc
     2401 aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt
     2461 aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct
     2521 gtcgattcga tactaacgcc gccatccagt ttaaacgagc tcgaattcat cgatgatatc
     2581 agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt gagtcgtatt
     2641 aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2701 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3061 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     3121 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3181 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3241 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3301 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3361 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3421 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3481 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3541 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3601 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3661 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3721 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3781 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3841 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3901 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3961 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4021 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4081 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4141 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4201 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4261 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4321 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4381 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4441 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4501 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4561 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4621 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4681 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgac
//
