LOCUS       sequence-62237-e        4888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            mApple with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4888)
  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 4888)
  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-yomApple-Kan
FEATURES             Location/Qualifiers
     source          1..4888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     84..103
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        184..202
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     184..201
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1003..1190
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1058..1082)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1265..2621
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1265..1608
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1609..2418
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1676..1695)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2424..2621
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2725..2744)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2726..2744)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2831..2848)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3002..3590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3082..3101)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3761..4552,4553..4621))
                     /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     4384..4403
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4622..4726)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4794..4812
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4850..4872)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
       61 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
      121 ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
      181 acgatttagg tgacactata gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg
      241 acggatcggt gacggtgctg gtttaattaa catggtatct aaaggtgagg aaaacaatat
      301 ggctatcatt aaagagttta tgcgtttcaa ggttcacatg gaaggttctg ttaatggtca
      361 tgagttcgaa atagaaggtg agggcgaagg gcgaccatac gaagcatttc aaactgctaa
      421 actcaaggta actaaaggcg gtcctcttcc atttgcctgg gatatattgt cacctcagtt
      481 catgtatggt tcaaaagttt acattaaaca tccagcggat atccctgact actttaaact
      541 gtctttccct gaaggtttta gatgggaaag agttatgaac ttcgaagatg gtggaattat
      601 tcacgtcaat caagattcaa gtttacagga tggcgtcttt atctacaagg tgaagttgag
      661 aggaacaaat tttccatccg atgggccagt tatgcaaaag aaaactatgg gctgggaagc
      721 ctctgaagag agaatgtacc cagaggacgg agccttaaaa agtgaaatca aaaagagatt
      781 gaagttgaag gatggtgggc actatgcagc agaagtgaaa acgacatata aggctaaaaa
      841 gcctgtccaa ctaccaggag cttatattgt ggacatcaaa cttgacatag taagccataa
      901 cgaagattac accatcgttg aacaatacga gagagctgaa ggtaggcatt ccacaggagg
      961 catggacgaa ctatacaaat aaggcgcgcc acttctaaat aagcgaattt cttatgattt
     1021 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
     1081 tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg
     1141 ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct
     1201 agggataaca gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc
     1261 cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc
     1321 atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc
     1381 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg
     1441 cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta
     1501 gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa
     1561 aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa
     1621 aagactcacg tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     1681 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
     1741 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     1801 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     1861 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca
     1921 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     1981 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     2041 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     2101 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg
     2161 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     2221 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     2281 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     2341 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     2401 ctcgatgagt ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt
     2461 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt
     2521 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa
     2581 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac
     2641 gccgccatcc agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct
     2701 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
     2761 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2821 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2881 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2941 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3001 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3061 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3121 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3181 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3241 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3301 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3361 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3421 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     3481 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3541 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3601 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3661 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3721 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3781 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3841 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3901 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3961 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4021 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4081 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4141 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4201 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4261 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4321 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4381 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4441 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4501 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4561 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4621 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4681 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4741 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4801 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     4861 tgcagctccc ggagacggtc acagcttg
//
