LOCUS       sequence-469816-        7284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing ymBaojin in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7284)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 7284)
  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: P415 ymBaojin
FEATURES             Location/Qualifiers
     source          1..7284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    52..73
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(88..93,94..111,112..118)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    126..142
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     131..153
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     150..166
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     150..166
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     185..205
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        187..205
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        223..621
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             631..1332
                     /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      1342..1589
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1358..1376)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1607..1626)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1608..1626)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1636..1653)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1636..1652)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1645..1667)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1793..2248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1880..1899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2090..2111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2549..2956
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2957..4051
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(4545..4564)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4664..4686
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4724..4742)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4784..5287
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5324..5428
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5429..5497,5498..6289)
                     /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     complement(5647..5666)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6460..7048
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6949..6968
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7202..7219
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
       61 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      121 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      181 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctcatagctt caaaatgttt
      241 ctactccttt tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa
      301 aacacccaag cacagcatac taaatttccc ctctttcttc ctctagggtg tcgttaatta
      361 cccgtactaa aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa
      421 aaaggcaata aaaattttta tcacgtttct ttttcttgaa aatttttttt ttgatttttt
      481 tctctttcga tgacctccca ttgatattta agttaataaa cggtcttcaa tttctcaagt
      541 ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca ttagaaagaa
      601 agcatagcaa tctaatctaa gttttctaga atggtttcta agggtgaaga agaaaacatg
      661 gcttctactc cattcaagtt ccaattgaag ggtactatta acggtaagtc tttcactgtt
      721 gaaggtgaag gtgaaggtaa ctctcatgaa ggttctcaca agggtaagta cgtttgtact
      781 tctggtaagt tgccaatgtc ttgggcagct ttgggtacta ctttcggtta cggtatgaag
      841 tactacacta agtacccatc tggtttaaag aactggttca gagaagttat gccaggtggt
      901 ttcacttacg atagacatat tcaatacaag ggtgatggtt ctattcatgc taagcatcaa
      961 catttcatga agaacggtac ttaccataac attgttgaat tcactggtca agatttcaag
     1021 gaaaattctc cagttttgac cggtgatatg aacgtttctt tgccaaatga agttccacaa
     1081 attccaagag atgatggtgt tgaatgtcca gttactttat tgtacccatt gttgtctgat
     1141 aagtctaagt atgttgaagc tcatcaatac actatttgta agccattgca taaccaacca
     1201 gctccagatg tcccatacca ttggattaga aagcaataca ctcaatctaa ggatgatgct
     1261 gaagaaagag atcatatttg tcaatctgaa actttagaag ctcatttgaa gggtatggat
     1321 gaattgtaca agtaactcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc
     1381 cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
     1441 tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     1501 tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg
     1561 ttttgggacg ctcgaaggct ttaatttgcg gccggtaccc aattcgccct atagtgagtc
     1621 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     1681 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     1741 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     1801 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1861 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1921 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1981 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2041 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2101 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2161 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2221 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     2281 gcggtatttc acaccgcata tcgaccggtc gaggagaact tctagtatat ctacatacct
     2341 aatattattg ccttattaaa aatggaatcc caacaattac atcaaaatcc acattctctt
     2401 caaaatcaat tgtcctgtac ttccttgttc atgtgtgttc aaaaacgtta tatttatagg
     2461 ataattatac tctatttctc aacaagtaat tggttgtttg gccgagcggt ctaaggcgcc
     2521 tgattcaaga aatatcttga ccgcagttaa ctgtgggaat actcaggtat cgtaagatgc
     2581 aagagttcga atctcttagc aaccattatt tttttcctca acataacgag aacacacagg
     2641 ggcgctatcg cacagaatca aattcgatga ctggaaattt tttgttaatt tcagaggtcg
     2701 cctgacgcat ataccttttt caactgaaaa attgggagaa aaaggaaagg tgagagcgcc
     2761 ggaaccggct tttcatatag aatagagaag cgttcatgac taaatgcttg catcacaata
     2821 cttgaagttg acaatattat ttaaggacct attgtttttt ccaataggtg gttagcaatc
     2881 gtcttacttt ctaacttttc ttacctttta catttcagca atatatatat atatatttca
     2941 aggatatacc attctaatgt ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt
     3001 tggtcaagaa atcacagccg aagccattaa ggttcttaaa gctatttctg atgttcgttc
     3061 caatgtcaag ttcgatttcg aaaatcattt aattggtggt gctgctatcg atgctacagg
     3121 tgttccactt ccagatgagg cgctggaagc ctccaagaag gctgatgccg ttttgttagg
     3181 tgctgtgggt ggtcctaaat ggggtaccgg tagtgttaga cctgaacaag gtttactaaa
     3241 aatccgtaaa gaacttcaat tgtacgccaa cttaagacca tgtaactttg catccgactc
     3301 tcttttagac ttatctccaa tcaagccaca atttgctaaa ggtactgact tcgttgttgt
     3361 cagagaatta gtgggaggta tttactttgg taagagaaag gaagacgatg gtgatggtgt
     3421 cgcttgggat agtgaacaat acaccgttcc agaagtgcaa agaatcacaa gaatggccgc
     3481 tttcatggcc ctacaacatg agccaccatt gcctatttgg tccttggata aagctaatgt
     3541 tttggcctct tcaagattat ggagaaaaac tgtggaggaa accatcaaga acgaattccc
     3601 tacattgaag gttcaacatc aattgattga ttctgccgcc atgatcctag ttaagaaccc
     3661 aacccaccta aatggtatta taatcaccag caacatgttt ggtgatatca tctccgatga
     3721 agcctccgtt atcccaggtt ccttgggttt gttgccatct gcgtccttgg cctctttgcc
     3781 agacaagaac accgcatttg gtttgtacga accatgccac ggttctgctc cagatttgcc
     3841 aaagaataag gtcaacccta tcgccactat cttgtctgct gcaatgatgt tgaaattgtc
     3901 attgaacttg cctgaagaag gtaaggccat tgaagatgca gttaaaaagg ttttggatgc
     3961 aggtatcaga actggtgatt taggtggttc caacagtacc accgaagtcg gtgatgctgt
     4021 cgccgaagaa gttaagaaaa tccttgctta aaaagattct ctttttttat gatatttgta
     4081 cataaacttt ataaatgaaa ttcataatag aaacgacacg aaattacaaa atggaatatg
     4141 ttcatagggt agacgaaact atatacgcaa tctacataca tttatcaaga aggagaaaaa
     4201 ggaggatgta aaggaataca ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa
     4261 aaagaattgc actttaacat taatattgac aaggaggagg gcaccacaca aaaagttagg
     4321 tgtaacagaa aatcatgaaa ctatgattcc taatttatat attggaggat tttctctaaa
     4381 aaaaaaaaaa tacaacaaat aaaaaacact caatgacctg accatttgat ggagtttaag
     4441 tcaatacctt cttgaaccat ttcccataat ggtgaaagtt ccctcaagaa ttttactctg
     4501 tcagaaacgg ccttaacgac gtagtcgata tggtgcactc tcagtacaat ctgctctgat
     4561 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4621 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4681 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4741 tttttatagg ttaatgtcat gataataatg gtttcttagg acggatcgct tgcctgtaac
     4801 ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg gaatttactc tgtgtttatt
     4861 tatttttatg ttttgtattt ggattttaga aagtaaataa agaaggtaga agagttacgg
     4921 aatgaagaaa aaaaaataaa caaaggttta aaaaatttca acaaaaagcg tactttacat
     4981 atatatttat tagacaagaa aagcagatta aatagatata cattcgatta acgataagta
     5041 aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc
     5101 ggcattaata cctgagagca ggaagagcaa gataaaaggt agtatttgtt ggcgatcccc
     5161 ctagagtctt ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac
     5221 tttctatttt taatttatat atttatatta aaaaatttaa attataatta tttttatagc
     5281 acgtgatgaa aaggacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5341 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5401 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5461 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5521 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5581 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5641 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5701 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5761 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5821 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5881 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5941 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     6001 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6061 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6121 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6181 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6241 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6301 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6361 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6421 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6481 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6541 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6601 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6661 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6721 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6781 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6841 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6901 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6961 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7021 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7081 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     7141 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     7201 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     7261 gcgttggccg attcattaat gcag
//
