LOCUS       sequence-489757-        4894 bp    DNA     circular SYN 20-OCT-2025
DEFINITION  Yeast tagging vector to create a gene fusion with yeast optimized 
            mBaojin with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4894)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4894)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pFA6a-link-ymBaojin-Kan
FEATURES             Location/Qualifiers
     source          1..4894
                     /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"
     CDS             491..1192
                     /codon_start=1
                     /product="bright and photostable monomeric green 
                     fluorescent protein derived from StayGold (Zhang et al., 
                     2024)"
                     /label=mBaoJin
                     /label=mStayGold
                     /note="/vntifkey=21"
                     /translation="MVSKGEEENMASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTTFGYGMKYYTKYPSGLKNWFREVMPGGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVPQIPRDDGVECPVTL
                     LYPLLSDKSKYVEAHQYTICKPLHNQPAPDVPYHWIRKQYTQSKDDAEERDHICQSETL
                     EAHLKGMDELYK"
     terminator      1228..1415
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1283..1307)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1490..2846
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1490..1833
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1834..2643
                     /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(1901..1920)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2649..2846
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2950..2969)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2951..2969)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3056..3073)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3227..3815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3307..3326)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3986..4777,4778..4846))
                     /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     4609..4628
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4847..4894,1..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 atggtttcta agggtgaaga agaaaacatg gcttctactc cattcaagtt
      541 ccaattgaag ggtactatta acggtaagtc tttcactgtt gaaggtgaag gtgaaggtaa
      601 ctctcatgaa ggttctcaca agggtaagta cgtttgtact tctggtaagt tgccaatgtc
      661 ttgggctgct ttgggtacta ctttcggtta cggtatgaag tactacacta agtacccatc
      721 tggtttaaag aactggttca gagaagttat gccaggtggt ttcacttacg atagacatat
      781 tcaatacaag ggtgatggtt ctattcatgc taagcatcaa catttcatga agaacggtac
      841 ttaccataac attgttgaat tcactggtca agatttcaag gaaaattctc cagttttgac
      901 cggtgatatg aacgtttctt tgccaaatga agttccacaa attccaagag atgatggtgt
      961 tgaatgtcca gttactttat tgtacccatt gttgtctgat aagtctaagt atgttgaagc
     1021 tcatcaatac actatttgta agccattgca taaccaacca gctccagatg tcccatacca
     1081 ttggattaga aagcaataca ctcaatctaa ggatgatgct gaagaaagag atcatatttg
     1141 tcaatctgaa actttagaag ctcatttgaa gggtatggat gaattgtaca agtaactcga
     1201 ggtcgacggc gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt
     1261 aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa
     1321 cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag
     1381 tatgaggtcg ctcttattga ccacacctct accggcagat ccgctaggga taacagggta
     1441 atatagatct gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc
     1501 ccagaatacc ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg
     1561 cccgtacatt tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg
     1621 ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc
     1681 tcccctcaca gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg
     1741 ttaggatttg ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata
     1801 cagttctcac atcacatccg aacataaaca accatgggta aggaaaagac tcacgtttcg
     1861 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     1921 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     1981 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     2041 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     2101 gatgatgcat ggttactcac cactgcgatc cccggcaaaa cagcattcca ggtattagaa
     2161 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     2221 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     2281 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     2341 ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat
     2401 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     2461 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     2521 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     2581 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     2641 taatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt
     2701 tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc
     2761 tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc
     2821 gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagttt
     2881 aaacgagctc gaattcatcg atgatatcag atccactagt ggcctatgcg gccgcggatc
     2941 tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat
     3001 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3061 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3121 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3181 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3241 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3301 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3361 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3421 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3481 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3541 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3601 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3661 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3721 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3781 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3841 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3901 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3961 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4021 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4081 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4141 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4201 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4261 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4321 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4381 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4441 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4501 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4561 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4621 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4681 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4741 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4801 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4861 aatattattg aagcatttat cagggttatt gtct
//
