LOCUS       sequence-62127-e        4840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized mKO2
            with CaUra3 selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4840)
  AUTHORS   Lee S, Lim WA, Thorn KS
  TITLE     Improved Blue, Green, and Red Fluorescent Protein Tagging Vectors 
            for S. cerevisiae.
  JOURNAL   PLoS One. 2013 Jul 2;8(7):e67902. doi: 10.1371/journal.pone.0067902.
            Print 2013.
  PUBMED    23844123
REFERENCE   2  (bases 1 to 4840)
  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: "PLoS One.";
            date: "2013-07-2"; volume: "8(7):e67902. doi"; pages: " 
            10.1371/journal.pone.0067902. Print 2013"
COMMENT     Sequence Label: pFA6a-link-yomKO2-CaURA3
FEATURES             Location/Qualifiers
     source          1..4840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      18..205
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(73..97)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        280..623
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1442..1639
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1746..1765)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1747..1765)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1852..1869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2023..2611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2103..2122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2782..3573,3574..3642))
                     /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     3405..3424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3643..3747)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3815..3833
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3871..3893)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3993..4012
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4093..4111
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4093..4110
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4181..4837
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
ORIGIN
        1 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
       61 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
      121 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
      181 ctcttattga ccacacctct accggcagat ccgctaggga taacagggta atatagatct
      241 gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc
      301 ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt
      361 tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg
      421 cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca
      481 gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg
      541 ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac
      601 atcacatccg aacataaaca accatgacag tcaacactaa gacctatagt gagagagcag
      661 aaactcatgc ctcaccagta gcacagcgat tatttcgatt aatggaactg aagaaaacca
      721 atttatgtgc atcaattgac gttgatacca ctaaggaatt ccttgaatta attgataaat
      781 taggtcctta tgtatgctta atcaagactc atattgatat aatcaatgat ttttcctatg
      841 aatccactat tgaaccatta ttagaacttt cacgtaaaca tcaatttatg atttttgaag
      901 atagaaaatt tgctgatatt ggtaataccg taaagaaaca atatattggt ggagtttata
      961 aaattagtag ttgggcagat attaccaatg ctcatggtgt cactgggaat ggagtggttg
     1021 aaggattaaa acagggagct aaagaaacca ccaccaacca agagccaaga gggttattga
     1081 tgttagctga attatcatca gtgggatcat tagcatatgg agaatattct caaaaaactg
     1141 ttgaaattgc taaatccgat aaggaatttg ttattggatt tattgcccaa cgtgatatgg
     1201 gtggccaaga agaaggattt gattggctta ttatgacacc tggagttgga ttagatgata
     1261 aaggtgatgg attaggacaa caatatagaa ctgttgatga agttgttagc actggaactg
     1321 atattatcat tgttggtaga ggattgtttg gtaaaggaag agatccagat attgaaggta
     1381 aaaggtatag aaatgctggt tggaatgctt atttgaaaaa gactggccaa ttataatcag
     1441 tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat
     1501 tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat
     1561 catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg
     1621 aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag tgtcgaaaac
     1681 gagctcgaat tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc
     1741 ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat
     1801 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1861 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1921 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1981 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2041 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2101 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2161 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     2221 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2281 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2341 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2401 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2461 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2521 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2581 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2641 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2701 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2761 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2821 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2881 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2941 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3001 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3061 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3121 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3181 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3241 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3301 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3361 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3421 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3481 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3541 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3601 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3661 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3721 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3781 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     3841 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     3901 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     3961 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     4021 gcaccatatg gacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat
     4081 catacacata cgatttaggt gacactatag aacgcggccg ccagctgaag cttcgtacgc
     4141 tgcaggtcga cggatcggtg acggtgctgg tttaattaac atggtgtccg taattaagcc
     4201 agaaatgaag atgaggtatt acatggatgg ttctgtcaat ggacacgagt ttactattga
     4261 aggtgagggt acgggcagac cttacgaagg ccatcaagag atgacattaa gagtaactat
     4321 ggctgaagga ggtccaatgc ctttcgcttt cgacttagtt tctcatgtgt tttgttacgg
     4381 gcatcgtgtg tttactaagt accctgaaga gataccagat tactttaaac aagccttccc
     4441 agagggactt tcatgggaaa gatcccttga atttgaagat ggagggagtg cttcagtttc
     4501 tgcacatatc agcttgagag gtaacacatt ttaccacaaa tcaaaattca caggcgtcaa
     4561 cttccctgcc gatgggccaa tcatgcaaaa tcaatctgtc gactgggaac catcaacaga
     4621 gaaaattaca gcatctgatg gtgttctcaa aggtgacgtt accatgtatt tgaaactaga
     4681 aggaggtggt aatcacaaat gccaaatgaa aacaacttat aaggccgcta aggaaatatt
     4741 ggaaatgcca ggcgaccatt acatcggtca tagactggtt agaaagaccg aaggcaatat
     4801 cactgaacag gttgaagatg cagtagcgca cagttaaggc
//
