LOCUS       sequence-452185-        5145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create a gene fusion with yeast optimized 
            miRFP680 with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5145)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 5145)
  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-Kan
FEATURES             Location/Qualifiers
     source          1..5145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     125..143
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(181..203)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     303..322
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        403..421
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     403..420
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1479..1666
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1534..1558)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1741..3097
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1741..2084
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2085..2894
                     /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(2152..2171)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2900..3097
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3201..3220)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3202..3220)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3307..3324)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3478..4066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3558..3577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4237..5028,5029..5097))
                     /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     4860..4879
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5098..5145,1..57))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
       61 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
      121 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
      181 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
      241 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
      301 tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg
      361 cggctacaat taatacataa ccttatgtat catacacata cgatttaggt gacactatag
      421 aacgcggccg ccagctgaag cttcgtacgc tgcaggtcga cggatcggtg acggtgctgg
      481 tttaattaac atggctgaag gttctgttgc tagacaacca gatttgttga cttgtgatga
      541 tgaaccaatt catattccag gtgctattca accacatggt ttgttattag ctttggctgc
      601 tgatatgacc attgttgctg gttctgataa cttgcctgaa ttgactggtt tagctattgg
      661 tgccttgatc ggtagatctg ctgctgatgt tttcgattct gaaacccata atagattgac
      721 tattgctttg gctgaaccag gtgctgctgt cggtgctcca attaccgtcg gtttcaccat
      781 gcgtaaggat gccggtttca ttggttcttg gcatagacat gatcaattaa ttttcttgga
      841 attggaacca ccacaacgtg acgttgccga accacaggct ttcttccgta gaaccaactc
      901 tgctattaga agattacaag ctgctgaaac tttggaatct gcttgtgctg ctgctgctca
      961 agaagttcgt aagattactg gtttcgatag agttatgatc tacagatttg cttctgattt
     1021 ctctggtgaa gttattgctg aagatagatg tgctgaagtt gaatctaagt tgggtttgca
     1081 ttacccagct tctactgttc cagctcaagc tagaagattg tacactatta atccagttag
     1141 aattattcca gatatcaact acagaccagt cccagttacc ccagatttga acccagttac
     1201 tggtagacct attgatttat ctttcgctat tttgagatct gtttccccat gtcatttgga
     1261 attcatgaga aacatcggta tgcatggtac catgtctatt tctattttga gaggtgaaag
     1321 attgtggggt ttgattgttt gtcatcatag aactccatac tacgttgact tggatggtag
     1381 acaagcctgt aagagagttg ctgaaagatt ggctacccaa attggtgtta tggaagaata
     1441 agcggccgcc tcgaggtcgg cgcgccactt ctaaataagc gaatttctta tgatttatga
     1501 tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt
     1561 aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt
     1621 ctcaggtata gtatgaggtc gctcttattg accacacctc taccggcaga tccgctaggg
     1681 ataacagggt aatatagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc
     1741 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
     1801 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
     1861 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag
     1921 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
     1981 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
     2041 ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt aaggaaaaga
     2101 ctcacgtttc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     2161 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
     2221 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     2281 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     2341 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc
     2401 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     2461 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     2521 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     2581 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat
     2641 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     2701 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     2761 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     2821 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     2881 atgagttttt ctaatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt
     2941 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat
     3001 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc
     3061 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg
     3121 ccatccagtt taaacgagct cgaattcatc gatgatatca gatccactag tggcctatgc
     3181 ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa
     3241 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3301 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3361 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3421 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3481 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3541 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3601 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3661 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3721 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3781 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3841 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3901 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3961 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4021 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4081 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4141 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4201 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4261 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4321 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4381 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4441 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4501 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4561 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4621 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4681 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4741 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4801 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4861 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4921 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4981 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5041 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5101 cttccttttt caatattatt gaagcattta tcagggttat tgtct
//
