LOCUS       sequence-61105-e        4678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with mKO1 with SpHis5 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4678)
  TITLE     Yeast tagging plasmids.
REFERENCE   2  (bases 1 to 4678)
  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-mKO1-SpHis5
FEATURES             Location/Qualifiers
     source          1..4678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             5..661
                     /codon_start=1
                     /product="monomeric Kusabira-Orange fluorescent protein 
                     (Karasawa et al., 2004)"
                     /label=mKO
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     terminator      682..869
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(737..761)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            944..2144
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        944..1287
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1288..1941
                     /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
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      1947..2144
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2248..2267)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2249..2267)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2354..2371)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2525..3113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2605..2624)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3284..4075,4076..4144))
                     /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     3907..3926
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4145..4249)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4317..4335
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4373..4395)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4495..4514
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4595..4613
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4595..4612
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 taacatggtg agtgtgatta aaccagagat gaagatgagg tactacatgg acggctccgt
       61 caatgggcat gagttcacaa ttgaaggtga aggcacaggc agaccttacg agggacatca
      121 agagatgaca ctacgcgtca caatggccaa gggcgggcca atgcctttcg cgtttgactt
      181 agtgtcacac gtgttctgtt acggccacag accttttact aaatatccag aagagatacc
      241 agactatttc aaacaagcat ttcctgaagg cctgtcatgg gaaaggtcgt tggagttcga
      301 agatggtggg tccgcttcag tcagtgcgca tataagcctt agaggaaaca ccttctacca
      361 caaatccaaa tttactgggg ttaactttcc tgccgatggt cctatcatgc aaaaccaaag
      421 tgttgattgg gagccatcaa ccgagaaaat tactgccagc gacggagttc tgaagggtga
      481 tgttacgatg tacctaaaac ttgaaggagg cggcaatcac aaatgccaat tcaagactac
      541 ttacaaggcg gcaaaaaaga ttcttaaaat gccaggaagc cattacatca gccatcgcct
      601 cgtcaggaaa accgaaggca acattactga gctggtagaa gatgcagtag ctcattcctg
      661 aggcgcgcca cttctaaata agcgaatttc ttatgattta tgatttttat tattaaataa
      721 gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa
      781 ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagtatgag
      841 gtcgctctta ttgaccacac ctctaccggc agatccgcta gggataacag ggtaatatag
      901 atctgtttag cttgcctcgt ccccgccggg tcacccggcc agcgacatgg aggcccagaa
      961 taccctcctt gacagtcttg acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta
     1021 catttagccc atacatcccc atgtataatc atttgcatcc atacattttg atggccgcac
     1081 ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc gagcagggaa acgctcccct
     1141 cacagacgcg ttgaattgtc cccacgccgc gcccctgtag agaaatataa aaggttagga
     1201 tttgccactg aggttcttct ttcatatact tccttttaaa atcttgctag gatacagttc
     1261 tcacatcaca tccgaacata aacaaccatg ggtaggaggg cttttgtaga aagaaatacg
     1321 aacgaaacga aaatcagcgt tgccatcgct ttggacaaag ctcccttacc tgaagagtcg
     1381 aattttattg atgaacttat aacttccaag catgcaaacc aaaagggaga acaagtaatc
     1441 caagtagaca cgggaattgg attcttggat cacatgtatc atgcactggc taaacatgca
     1501 ggctggagct tacgacttta ctcaagaggt gatttaatca tcgatgatca tcacactgca
     1561 gaagatactg ctattgcact tggtattgca ttcaagcagg ctatgggtaa ctttgccggc
     1621 gttaaaagat ttggacatgc ttattgtcca cttgacgaag ctctttctag aagcgtagtt
     1681 gacttgtcgg gacggcccta tgctgttatc gatttgggat taaagcgtga aaaggttggg
     1741 gaattgtcct gtgaaatgat ccctcactta ctatattcct tttcggtagc agctggaatt
     1801 actttgcatg ttacctgctt atatggtagt aatgaccatc atcgtgctga aagcgctttt
     1861 aaatctctgg ctgttgccat gcgcgcggct actagtctta ctggaagttc tgaagtccca
     1921 agcacgaagg gagtgttgta aagagtactg acaataaaaa gattcttgtt ttcaagaact
     1981 tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga
     2041 tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag
     2101 taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact
     2161 aacgccgcca tccagtttaa acgagctcga attcatcgat gatatcagat ccactagtgg
     2221 cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc
     2281 aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2341 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2401 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2461 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2521 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2581 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2641 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2701 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     2761 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2821 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2881 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2941 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     3001 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3061 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3121 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3181 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3241 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3301 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3361 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3421 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3481 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3541 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3601 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3661 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3721 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3781 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3841 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3901 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3961 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4021 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4081 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4141 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4201 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4261 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     4321 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     4381 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     4441 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     4501 cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa cgcggctaca
     4561 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaacgcggc
     4621 cgccagctga agcttcgtac gctgcaggtc gacggatcgg tgacggtgct ggtttaat
//
