LOCUS       sequence-452410-        4743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create a gene fusion with yeast optimized 
            mBaojin with SpHis5 selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4743)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4743)
  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-ymBaojin-SpHis5
FEATURES             Location/Qualifiers
     source          1..4743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     327..346
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(565..669)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     737..755
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(793..815)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     915..934
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1015..1033
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1015..1032
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1103..1804
                     /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      1845..2032
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1900..1924)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            2107..3307
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        2107..2450
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2451..3104
                     /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="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHTNQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMSNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      3110..3307
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3411..3430)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3412..3430)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3517..3534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3688..4276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3768..3787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4447..4743,1..495,496..564))
                     /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"
ORIGIN
        1 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
       61 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
      121 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
      181 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
      241 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
      301 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
      361 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
      421 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
      481 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
      541 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
      601 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
      661 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
      721 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
      781 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
      841 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
      901 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt
      961 gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag
     1021 gtgacactat agaacgcggc cgccagctga agcttcgtac gctgcaggtc gacggatcgg
     1081 tgacggtgct ggtttaatta acatggtttc taagggtgaa gaagaaaaca tggcttctac
     1141 tccattcaag ttccaattga agggtactat taacggtaag tctttcactg ttgaaggtga
     1201 aggtgaaggt aactctcatg aaggttctca caagggtaag tacgtttgta cttctggtaa
     1261 gttgccaatg tcttgggctg ctttgggtac tactttcggt tacggtatga agtactacac
     1321 taagtaccca tctggtttaa agaactggtt cagagaagtt atgccaggtg gtttcactta
     1381 cgatagacat attcaataca agggtgatgg ttctattcat gctaagcatc aacatttcat
     1441 gaagaacggt acttaccata acattgttga attcactggt caagatttca aggaaaattc
     1501 tccagttttg accggtgata tgaacgtttc tttgccaaat gaagttccac aaattccaag
     1561 agatgatggt gttgaatgtc cagttacttt attgtaccca ttgttgtctg ataagtctaa
     1621 gtatgttgaa gctcatcaat acactatttg taagccattg cataaccaac cagctccaga
     1681 tgtcccatac cattggatta gaaagcaata cactcaatct aaggatgatg ctgaagaaag
     1741 agatcatatt tgtcaatctg aaactttaga agctcatttg aagggtatgg atgaattgta
     1801 caagtaagcg gccgcctcga ggtcggcgcg ccacttctaa ataagcgaat ttcttatgat
     1861 ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg
     1921 actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt
     1981 tgctttctca ggtatagtat gaggtcgctc ttattgacca cacctctacc ggcagatccg
     2041 ctagggataa cagggtaata tagatctgtt tagcttgcct cgtccccgcc gggtcacccg
     2101 gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg
     2161 gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca
     2221 tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc
     2281 tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg
     2341 tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt
     2401 aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc atgggtagga
     2461 gggcttttgt agaaagaaat acgaacgaaa cgaaaatcag cgttgccatc gctttggaca
     2521 aagctccctt acctgaagag tcgaatttta ttgatgaact tataacttcc aagcatacaa
     2581 accaaaaggg agaacaagta atccaagtag acacgggaat tggattcttg gatcacatgt
     2641 atcatgcact ggctaaacat gcaggctgga gcttacgact ttactcaaga ggtgatttaa
     2701 tcatcgatga tcatcacact gcagaagata ctgctattgc acttggtatt gcattcaagc
     2761 aggctatgag taactttgcc ggcgttaaaa gatttggaca tgcttattgt ccacttgacg
     2821 aagctctttc tagaagcgta gttgacttgt cgggacggcc ctatgctgtt atcgatttgg
     2881 gattaaagcg tgaaaaggtt ggggaattgt cctgtgaaat gatccctcac ttactatatt
     2941 ccttttcggt agcagctgga attactttgc atgttacctg cttatatggt agtaatgacc
     3001 atcatcgtgc tgaaagcgct tttaaatctc tggctgttgc catgcgcgcg gctactagtc
     3061 ttactggaag ttctgaagtc ccaagcacga agggagtgtt gtaaagagta ctgacaataa
     3121 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     3181 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     3241 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     3301 tatactgctg tcgattcgat actaacgccg ccatccagtt taaacgagct cgaattcatc
     3361 gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg
     3421 agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg
     3481 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3541 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3601 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3661 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3721 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3781 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3841 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3901 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3961 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4021 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4081 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4141 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4201 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4261 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4321 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4381 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4441 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4501 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4561 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4621 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4681 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4741 gcg
//
