LOCUS       sequence-292100-        7927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP-SED5 as an additional copy in genome, use 
            auxotrophic marker URA3(Kluyveromyces lactis). Marker for yeast 
            early golgi..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7927)
  AUTHORS   Zhu J, Zhang ZT, Tang SW, Zhao BS, Li H, Song JZ, Li D, Xie Z
  TITLE     A Validated Set of Fluorescent-Protein-Based Markers for Major 
            Organelles in Yeast (Saccharomyces cerevisiae).
  JOURNAL   MBio. 2019 Sep 3;10(5). pii: mBio.01691-19. doi: 
            10.1128/mBio.01691-19.
  PUBMED    31481383
REFERENCE   2  (bases 1 to 7927)
  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"; journalName: "MBio."; 
            date: "2019-09-3"; volume: "10(5). pii: mBio.01691-19. doi"; pages: 
            " 10.1128/mBio.01691-19"
COMMENT     Sequence Label: ZJOM1
FEATURES             Location/Qualifiers
     source          1..7927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..217
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     protein_bind    complement(1161..1194)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2234..2947
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2234..2947
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     promoter        complement(4688..4706)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4727..4743)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4727..4743)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4740..4762)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4751..4767
                     /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)."
     promoter        complement(join(4775..4781,4782..4799,4800..4805))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4820..4841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4958..4975)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5129..5717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5209..5228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5888..6679,6680..6748))
                     /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     6511..6530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6749..6853)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6879..7334)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7016..7037)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7228..7247
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7461..7483
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7475..7492
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7476..7492
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7499..7518
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7499..7517
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttcaattcat cttttttttt tttgttcttt tttttgattc cggtttcttt gaaatttttt
       61 tgattcggta atctccgagc agaaggaaga acgaaggaag gagcacagac ttagattggt
      121 atatatacgc atatgtggtg ttgaagaaac atgaaattgc ccagtattct taacccaact
      181 gcacagaaca aaaacctgca ggaaacgaag ataaatcctc gagatgtcca caaaatcata
      241 taccagtaga gctgagactc atgcaagtcc ggttgcatcg aaacttttac gtttaatgga
      301 tgaaaagaag accaatttgt gtgcttctct tgacgttcgt tcgactgatg agctattgaa
      361 acttgttgaa acgttgggtc catacatttg ccttttgaaa acacacgttg atatcttgga
      421 tgatttcagt tatgagggta ctgtcgttcc attgaaagca ttggcagaga aatacaagtt
      481 cttgatattt gaggacagaa aattcgccga tatcggtaac acagtcaaat tacaatatac
      541 atcgggcgtt taccgtatcg cagaatggtc tgatatcacc aacgcccacg gggttactgg
      601 tgctggtatt gttgctggct tgaaacaagg tgcgcaagag gtcaccaaag aaccaagggg
      661 attattgatg cttgctgaat tgtcttccaa gggttctcta gcacacggtg aatatactaa
      721 gggtaccgtt gatattgcaa agagtgataa agatttcgtt attgggttca ttgctcagaa
      781 cgatatggga ggaagagaag aagggtttga ttggctaatc atgaccccag gtgtaggttt
      841 agacgacaaa ggcgatgcat tgggtcagca gtacagaacc gtcgacgaag ttgtaagtgg
      901 tggatcagat atcatcattg ttggcagagg acttttcgcc aagggtagag atcctaaggt
      961 tgaaggtgaa agatacagaa atgctggatg ggaagcgtac caaaagagaa tcagcgctcc
     1021 ccattaatta tacaggaaac ttaatagaac aaatcacata tttaatctaa tagccacctg
     1081 cattggcacg gtgcaacact acttcaactt catcttacaa aaagatcacg tgatctgttg
     1141 tattgggatc tctagaccta ataacttcgt atagcataca ttatacgaag ttatattaag
     1201 ggttgtcgac ctgcagcgta catcgataag cttgatatcg aattcctgca gcccggggga
     1261 tccttgacac cgcctaccaa gggtttgcca ctggagattt ggacaaggat gcctatgctg
     1321 tgcgtctagg tgtggagaag ctttcaacgg tctctcccgt ctttgtctgt cagtcctttg
     1381 ccaagaacgc cggtatgtac ggtgagcgtg taggttgttt ccatctagca cttacaaaac
     1441 aagctcaaaa caaaactata aagcctgctg ttacatctca attggccaaa atcattcgta
     1501 gtgaagtgtc caacccaccc gcctacggcg ctaagattgt cgctaaactg ttggaaacgc
     1561 cagaattaac ggaacagtgg cacaaggata tggttaccat gtcctccaga attacgaaaa
     1621 tgaggcacgc attaagagac catttagtca agttgggcac tcctggcaac tgggatcata
     1681 tagtaaatca atgcgggatg ttctccttta caggattgac tcctcaaatg gttaaacgac
     1741 ttgaagaaac ccacgcagtt tacttggttg cctcaggtag agcttctatt gctggattga
     1801 atcaaggaaa cgtggaatac gtggctaaag ccattgatga agtggtgcgc ttctatacta
     1861 ttgaagctaa attgtaatga ttaaataata cttatactta cctattacac tcttcatatt
     1921 ttttttaatg taaagaaagt aaaaaaaaaa aaaaataaaa aaaagagaaa tgctctttct
     1981 ttaccttcac aggtgtatta ttatgttgtt gtaatatccg ggtaggttta ttgatcgagg
     2041 cgaggcgaaa gggcatattc gaaacaatca ttatgataat aatgtcaata tagaaattga
     2101 aaaaaaaaaa aaaatcctac cgccgcatcg acgctcctgt tagacctttg aacggaaatc
     2161 acccactctt tactgtatat attttattcc acctctgttt ccaacgacca tttcaccata
     2221 ccacacaact cccatgtcta aaggtgaaga attattcact ggtgttgtcc caattttggt
     2281 tgaattagat ggtgatgtta atggtcacaa attttctgtc tccggtgaag gtgaaggtga
     2341 tgctacttac ggtaaattga ccttaaaatt tatttgtact actggtaaat tgccagttcc
     2401 atggccaacc ttagtcacta ctttaactta tggtgttcaa tgtttttcta gatacccaga
     2461 tcatatgaaa caacatgact ttttcaagtc tgccatgcca gaaggttatg ttcaagaaag
     2521 aactattttt ttcaaagatg acggtaacta caagaccaga gctgaagtca agtttgaagg
     2581 tgatacctta gttaatagaa tcgaattaaa aggtattgat tttaaagaag atggtaacat
     2641 tttaggtcac aaattggaat acaactataa ctctcacaat gtttacatca tggctgacaa
     2701 acaaaagaat ggtatcaaag ttaacttcaa aattagacac aacattgaag atggttctgt
     2761 tcaattagct gaccattatc aacaaaatac tccaattggt gatggtccag tcttgttacc
     2821 agacaaccat tacttatcca ctcaatctgc cttatccaaa gatccaaacg aaaagagaga
     2881 ccacatggtc ttgttagaat ttgttactgc tgctggtatt acccatggta tggatgaatt
     2941 gtacaaagct tcgtacgctg caggtcgacg gatccccggg ttaattaaca tgaacataaa
     3001 ggatagaact tcagaatttc aacaaagtgt gttgagttac aagaaaagaa acaaaaactt
     3061 tagagaacag cagagggaac gtctacaaga gaaagaaagt gaaaactttg ccaacaacac
     3121 aaccggcaat gggaaaagtg tctctgagtt tcaaaaaaag gcttctggca tagctcacga
     3181 aatttcatcc accgcacaat tattatcgaa attagcagtg ctggccaaaa ggaaaccgat
     3241 gttcaatgac aaccctgtag agattgccga gctatcattt ttgatcaaaa ggaagattta
     3301 tgccatcgaa caaagtttag tgcaattaag tcagctcaag aaaaccgacg tgaatggcaa
     3361 cacatcaaat caatcttcta agcaacctag tgccgtacag cactccaaaa atgtcgtaaa
     3421 tcttttaaac acgcaaatga aaaacatatc aggaagtttt aaagacgtat tggaggaaag
     3481 gcaacgacta gaaatggcta acaaagacag atggcaaaaa ctaaccaccg acactggaca
     3541 tgcaccagct gatgaccaaa cgcaaagcaa tcacgccgcg gacttgacca cgtataacaa
     3601 ctccaaccca ttcatgacct cactactaga tgaatcctca gaaaagaata acaactcgtc
     3661 caaccaagga gaactgtctt tccctcaaaa cgactctcaa ctaatgttaa tggaagaggg
     3721 acagttatcc aataacgtct acttacaaga aagaaatagg gcggtggaga caatagaatc
     3781 cacaatccag gaagtgggaa atctcttcca acaactggcc tccatggttc aagaacaggg
     3841 cgaagtaatc caaaggattg atgcaaacgt agacgacatc gatttaaaca ttagtggtgc
     3901 tcaaagggaa cttttgaaat acttcgacag gataaagagt aatagatggt tagccgcaaa
     3961 ggtttttttt ataatctttg tatttttcgt tatttgggtt ttagtcaatt aagcggccgc
     4021 aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt
     4081 aattatatca gttattaccc gggaatctcg gtcgtaatga tttctataat gacgaaaaaa
     4141 aaaaaattgg aaagaaaaag cttcatggcc tttataaaaa ggaactatcc aatacctcgc
     4201 cagaaccaag taacagtatt ttacggggca caaatcaaga acaataagac aggactgtaa
     4261 agatggacgc attgaactcc aaagaacaac aagagttcca aaaagtagtg gaacaaaagc
     4321 aaatgaagga tttcatgcgt ttgtactcta atctggtaga aagatgtttc acagactgtg
     4381 tcaatgactt cacaacatca aagctaacca ataaggaaca aacatgcatc atgaagtgct
     4441 cagaaaagtt cttgaagcat agcgaacgtg tagggcagcg tttccaagaa caaaacgctg
     4501 ccttgggaca aggcttgggc cgataaggtg tactggcgta tatatatcta attatgtatc
     4561 tctggtgtag cccattttta gcatgtaaat ataaagagaa accatatcta atctaaccaa
     4621 atccaaacaa aattcaatag ttacgtagac gtcctacgat tccgcggtgg agctccagct
     4681 tttgttccct ttagtgaggg ttaatttcga gcttggcgta atcatggtca tagctgtttc
     4741 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4801 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4861 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4921 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4981 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5041 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5101 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5161 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5221 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5281 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5341 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5401 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5461 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5521 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5581 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5641 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5701 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5761 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5821 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5881 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5941 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6001 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6061 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6121 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6181 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6241 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6301 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6361 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6421 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6481 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6541 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6601 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6661 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6721 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6781 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6841 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     6901 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     6961 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     7021 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     7081 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     7141 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     7201 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     7261 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     7321 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     7381 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     7441 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta
     7501 atacgactca ctatagggcg aattgggtac ccgttgctaa attcgagtga aacacaggaa
     7561 gaccagaaaa tcctcatttc atccatatta acaataattt caaatgttta tttgcattat
     7621 ttgaaactag ggaagacaag caacgaaacg tttttgaaaa ttttgagtat tttcaataaa
     7681 tttgtagagg actcagatat tgaaaaaaag ctacagcaat taatacttga taagaagagt
     7741 attgagaagg gcaacggttc atcatctcat ggatctgcac atgaacaaac accagagtca
     7801 aacgacgttg aaattgaggc tactgcgcca attgaggcct tttttgcgag gcatatttat
     7861 ggtgaaggat aagttttgac catcaaagaa ggttaatgtg gctgtggttt cagggtccat
     7921 aaagctt
//
