LOCUS       sequence-456001-        7672 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 7672)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 7672)
  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: pDRF1 ymBaojin
FEATURES             Location/Qualifiers
     source          1..7672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    800..824
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             843..1544
                     /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"
     protein_bind    complement(1562..1586)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1648..1667)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1649..1667)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1815..2002
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1870..1894)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2014..2031)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2014..2030)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2023..2045)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2466..3812
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3923..4726)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4727..4942)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4967..4985)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5053..5157
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5158..5226,5227..6018)
                     /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(5376..5395)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6189..6777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6678..6697
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6931..6948
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7065..7086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7101..7106,7107..7124,7125..7131)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7139..7155
                     /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     7144..7166
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7163..7179
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7163..7179
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7209..7664)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7346..7367)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7558..7577
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caagcttcct gaaacggaga aacataaaca ggcattgctg ggatcaccca tacatcactc
       61 tgttttgcct gaccttttcc ggtaatttga aaacaaaccc ggtctcgaag cggagatccg
      121 gcgataatta ccgcagaaat aaacccatac acgagacgta gaaccagccg cacatggccg
      181 gagaaactcc tgcgagaatt tcgtaaactc gcgcgcattg catctgtatt tcctaatgcg
      241 gcacttccag gcctcgatcg agaccgttta tccattgctt ttttgttgtc tttttccctc
      301 gttcacagaa agtctgaaga agctatagta gaactatgag ctttttttgt ttctgttttc
      361 cttttttttt tttttacctc tgtggaaatt gttactctca cactctttag ttcgtttgtt
      421 tgttttgttt attccaatta tgaccggtga cgaaacgtgg tcgatggtgg gtaccgctta
      481 tgctcccctc cattagtttc gattatataa aaaggccaaa tattgtatta ttttcaaatg
      541 tcctatcatt atcgtctaac atctaatttc tcttaaattt tttctctttc tttcctataa
      601 caccaatagt gaaaatcttt ttttcttcta tatctacaaa aacttttttt ttctatcaac
      661 ctcgttgata aattttttct ttaacaatcg ttaataatta attaattgga aaataaccat
      721 tttttctctc ttttatacac acattcaaaa gaaagaaaaa aaatataccc cagcctcgac
      781 tagtgaattc gcggccatca caagtttgta caaaaaagca ggctgctagc ccatggtcta
      841 gaatggtttc taagggtgaa gaagaaaaca tggcttctac tccattcaag ttccaattga
      901 agggtactat taacggtaag tctttcactg ttgaaggtga aggtgaaggt aactctcatg
      961 aaggttctca caagggtaag tacgtttgta cttctggtaa gttgccaatg tcttgggctg
     1021 ctttgggtac tactttcggt tacggtatga agtactacac taagtaccca tctggtttaa
     1081 agaactggtt cagagaagtt atgccaggtg gtttcactta cgatagacat attcaataca
     1141 agggtgatgg ttctattcat gctaagcatc aacatttcat gaagaacggt acttaccata
     1201 acattgttga attcactggt caagatttca aggaaaattc tccagttttg accggtgata
     1261 tgaacgtttc tttgccaaat gaagttccac aaattccaag agatgatggt gttgaatgtc
     1321 cagttacttt attgtaccca ttgttgtctg ataagtctaa gtatgttgaa gctcatcaat
     1381 acactatttg taagccattg cataaccaac cagctccaga tgtcccatac cattggatta
     1441 gaaagcaata cactcaatct aaggatgatg ctgaagaaag agatcatatt tgtcaatctg
     1501 aaactttaga agctcatttg aagggtatgg atgaattgta caagtaactc gaggcggccg
     1561 cacccagctt tcttgtacaa agtggtgtag ttaattcatg atggccgctg caggtcgacc
     1621 tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacgc gcggatccag
     1681 ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc ggcttgtcta
     1741 ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga tacgttgttg
     1801 acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     1861 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     1921 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc
     1981 ttattgacca cacctctacc ggcatgccaa ttcactggcc gtcgttttac aacgtcgtga
     2041 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2101 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2161 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2221 cataatcgga tcgtacttgt tacccatcat tgaattttga acatccgaac ctgggagttt
     2281 tccctgaaac agatagtata tttgaacctg tataataata tatagtctag cgctttacgg
     2341 aagacaatgt atgtatttcg gttcctggag aaactattgc atctattgca taggtaatct
     2401 tgcacgtcgc atccccggtt cattttctgc gtttccatct tgcacttcaa tagcatatct
     2461 ttgttaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa
     2521 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc
     2581 tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc
     2641 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag
     2701 agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg
     2761 agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta
     2821 taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac
     2881 tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat
     2941 tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt atattctata
     3001 ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg
     3061 gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt
     3121 ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt
     3181 ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt
     3241 caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa
     3301 agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta
     3361 cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa
     3421 agcgctctga agttcctata ctttctagct agagaatagg aacttcggaa taggaacttc
     3481 aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca catacagctc
     3541 actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca tgagaagaac
     3601 ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc gtctatttat gtaggatgaa
     3661 aggtagtcta gtacctcctg tgatattatc ccattccatg cggggtatcg tatgcttcct
     3721 tcagcactac cctttagctg ttctatatgc tgccactcct caattggatt agtctcatcc
     3781 ttcaatgcta tcatttcctt tgatattgga tcgatccgat gataagctgt caaacatgag
     3841 aattgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt agtatacatg
     3901 catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa tatgcttccc
     3961 agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt gcaaatagtc
     4021 ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg gttctatact
     4081 gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata atcaaccaat
     4141 cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata acaaaatctt
     4201 tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat agggagccct
     4261 tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact tcttctgccg
     4321 cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc gtaatgtctg
     4381 cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta ttaccaatgt
     4441 cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat gcctttagcg
     4501 gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac
     4561 aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta cgaacatcca
     4621 atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg gcagcaacag
     4681 gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt tatcttcgtt
     4741 tcctgcatgt ttttgttctg tgcagttggg ttaagaatac tgggcaattt catgtttctt
     4801 caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct
     4861 tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat
     4921 aaaaaaaaaa tgatgaattg aaaagctaat tcttgaagac gaaagggcct cgtgatacgc
     4981 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5041 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5101 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5161 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5221 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5281 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5341 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5401 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5461 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5521 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5581 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5641 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5701 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5761 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5821 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5881 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5941 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6001 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6061 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6121 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6181 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6241 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6301 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6361 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6421 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6481 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6541 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6601 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6661 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6721 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6781 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6841 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6901 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6961 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7021 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7081 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7141 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttacc
     7201 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     7261 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     7321 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     7381 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     7441 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     7501 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     7561 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     7621 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gc
//
