LOCUS       sequence-265897-        8982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of EMC1-2GFP as an additional copy in genome, use 
            auxotrophic marker URA3(Candida albicans). Marker for yeast 
            endoplasmic reticulum (ER)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8982)
  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 8982)
  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: ZJOM22
FEATURES             Location/Qualifiers
     source          1..8982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3343..4056
                     /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
                     FVTAAGITHGIDELYK"
     CDS             4093..4809
                     /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"
     CDS             4093..4806
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      4830..5017
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4885..4909)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    5300..5333
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      6288..6485
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    6539..6572
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(6626..6645)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6627..6645)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6732..6749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6903..7491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6983..7002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7662..8453,8454..8522))
                     /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     8285..8304
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8523..8627)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8695..8713
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8751..8773)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8873..8892
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(8973..8982,1..9)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(8973..8982,1..8)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 cactatagaa cgcggccgcc agctgaagct tgctactttc tcagtcactg ttggagctta
       61 ctcgcctgaa ctttcttaaa tacgacccct ggtgcgacga aatgaaatcc gtacgtgaca
      121 agaagttttt caacaatatt gtcaaatatt atcacgaggg catagacccc accaaaataa
      181 aagtcaagga tgctatgaac ggtactcctc tctcgacaaa tatccctgag gtcaaacaaa
      241 gcgtcgctct cgctagagcg caagttgagg cgcagaatcc cattattaaa tggttcggac
      301 ccttggaata caagcccatg tctttcaacg agtacctcaa tcgcatggaa tttcacttgg
      361 acatgttcga gttttttcaa aataaaagaa acattagaga aaattccatt gaactcatca
      421 attccatatc ccacaatccg cagtcttctt ctactggcct tgaaggtctt tccgagtcca
      481 aaaaactcca tctacaaaat gtggaaaaaa gactgcattt cttagcatct tcgggagatt
      541 ccatttccgc accagtaaag aagagatcca gcaccacact ctcccgaggt gtcattttgc
      601 cccatgacac gaaaggcccg caagatattg atctcgatac aataagatcg ctttatgatc
      661 catggatgac tttggcctta gaaacttcgc taagcatcaa attcatacca actaccatgc
      721 cctcccatac caagacaccc actagcacgg accagccgtt accagggcct acccccaagg
      781 ctctcactaa tgaaaagaca cattaggtct ttgcggattg cttagttgta ccgtgcgtat
      841 attaaatcct tcttcgtaca ccgtgatttt tcagacggct accagagaaa atatggctgg
      901 gatttcatat tatatatgca aatatataat aacatatata tatatacact ccattaataa
      961 ctaataccta cattcatatc aacactttca tcctcccctt cccccatagc aatgaagata
     1021 acgtgtacag acttggtgta cgtcttcatt ttactcttcc taaacacgag ttgtgtccaa
     1081 gccgtttttt cagatgatgc atttatcact gattggcaac tggctaactt aggtccttgg
     1141 gagaaagtca tccctgattc tcgagaccgc aacagggttc tcatcttatc gaaccctacc
     1201 gaaacttcct gcttagtttc ttcgtttaac gtttcttccg gacagattct tttcagaaac
     1261 gttttaccct ttaccattga tgagattcaa ctggatagta atgaccataa cgcaatggtt
     1321 tgtgtgaact cttcaagcaa ccattggcag aaatatgatt tacacgattg gtttttacta
     1381 gaggaaggcg tagataatgc cccttctacg accattttac ctcaatcctc atatttaaac
     1441 gatcaagtat ctattaagaa caatgaacta catattctcg atgagcagtc aaaactggca
     1501 gaatggaaat tggagttacc tcaagggttc aataaagtgg aatattttca tcgtgaagat
     1561 cccctggcgt tagtgttgaa cgttaatgat acccaatata tgggattctc tgccaatggc
     1621 acagaattga tccccgtttg gcaaagagat gaatggttga ctaacgtggt agactatgct
     1681 gtattggacg tcttcgattc tagggatgtg gagttgaaca aagatatgaa agcggaactt
     1741 gattcaaatt cgctttggaa tgcttactgg cttagattga caactaattg gaatcgcctt
     1801 atcaacttat tgaaagaaaa ccaattctca ccaggacgtg tcttcactaa actcctagct
     1861 ctagacgcta aggataccac ggtatcagat ttgaagttcg gattcgccaa aatcttaatt
     1921 gttttgacgc atgatggctt tatcggcggc cttgatatgg tcaataaggg ccaacttatc
     1981 tggaaactcg atttagaaat tgatcagggc gtcaaaatgt tctggacgga taaaaaccat
     2041 gacgaacttg ttgttttttc gcatgatggg cattatttga caattgaagt tactaaagat
     2101 caaccgatta tcaaatcaag atccccccta tctgaaagga aaactgttga ttccgttatt
     2161 aggctgaatg aacatgatca ccagtatctg attaagtttg aggataagga tcatttactg
     2221 ttcaaattga atcccggcaa gaatacggat gtaccaatag ttgccaacaa ccattctagt
     2281 tcccacatat tcgtcacaga gcatgacacg aatggcattt atggctacat aatcgaaaac
     2341 gatacggtaa aacaaacttg gaaaaaagcc gtaaattcga aagagaaaat ggtggcatat
     2401 agcaagaggg aaacaacaaa cctaaacact cttggtatta cactaggtga caaatcggtt
     2461 ctttataaat atttgtaccc caacctagcg gcttatctga tcgctaatga agaacatcat
     2521 acaatcactt ttaacttaat tgataccatt acaggagaaa tcctcattac ccaagagcac
     2581 aaggattctc cggattttag gtttccaatg gatattgttt tcggtgaata ttgggtcgtt
     2641 tattcctatt tcagttctga acctgttcca gaacaaaagt tagtagtggt ggaattatat
     2701 gagtcactaa ccccagatga gcgtttgtct aactcaagcg acaatttttc ttatgatcca
     2761 ttgactggac acattaacaa acctcaattt caaactaaac aattcatttt tcccgagatt
     2821 atcaaaacaa tgtccatttc caagacaacg gatgatatta ccacaaaggc aatcgttatg
     2881 gaattagaaa atggacaaat cacctacata ccaaagcttt tattgaatgc aagaggtaaa
     2941 ccagcagaag aaatggccaa ggataagaaa aaagagttta tggctacccc atacacgcca
     3001 gttatcccaa ttaatgataa tttcattatc actcatttca gaaatctatt gccaggatcc
     3061 gattcgcagt tgatctccat cccaaccaat ctggaatcca caagcattat atgtgatcta
     3121 ggccttgatg tattttgtac aaggatcaca ccttcgggcc aatttgattt aatgagtcct
     3181 actttcgaaa agggtaaatt gcttattact atattcgtct tgttggtgat cacgtatttt
     3241 atccgtcctt ctgtttcaaa caagaagttg aaatcccaat ggctaattaa agcttcgtac
     3301 gctgcaggtc gacggatcgg tgacggtgct ggtttaatta acatgtctaa aggtgaagaa
     3361 ttattcactg gtgttgtccc aattttggtt gaattagatg gtgatgttaa tggtcacaaa
     3421 ttttctgtct ccggtgaagg tgaaggtgat gctacttacg gtaaattgac cttaaaattt
     3481 atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttaacttat
     3541 ggtgttcaat gtttttctag atacccagat catatgaaac aacatgactt tttcaagtct
     3601 gccatgccag aaggttatgt tcaagaaaga actatttttt tcaaagatga cggtaactac
     3661 aagaccagag ctgaagtcaa gtttgaaggt gataccttag ttaatagaat cgaattaaaa
     3721 ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca aattggaata caactataac
     3781 tctcacaatg tttacatcat ggctgacaaa caaaagaatg gtatcaaagt taacttcaaa
     3841 attagacaca acattgaaga tggttctgtt caattagctg accattatca acaaaatact
     3901 ccaattggtg atggtccagt cttgttacca gacaaccatt acttatccac tcaatctgcc
     3961 ttatccaaag atccaaacga aaagagagac cacatggtct tgttagaatt tgttactgct
     4021 gctggtatta cccatggtat tgatgaattg tacaaaattc cagttccaac gcgtgatcta
     4081 gagaagccat tcatgtctaa aggtgaagaa ttattcactg gtgttgtccc aattttggtt
     4141 gaattagatg gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat
     4201 gctacttacg gtaaattgac cttaaaattt atttgtacta ctggtaaatt gccagttcca
     4261 tggccaacct tagtcactac tttaacttat ggtgttcaat gtttttctag atacccagat
     4321 catatgaaac aacatgactt tttcaagtct gccatgccag aaggttatgt tcaagaaaga
     4381 actatttttt tcaaagatga cggtaactac aagaccagag ctgaagtcaa gtttgaaggt
     4441 gataccttag ttaatagaat cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt
     4501 ttaggtcaca aattggaata caactataac tctcacaatg tttacatcat ggctgacaaa
     4561 caaaagaatg gtatcaaagt taacttcaaa attagacaca acattgaaga tggttctgtt
     4621 caattagctg accattatca acaaaatact ccaattggtg atggtccagt cttgttacca
     4681 gacaaccatt acttatccac tcaatctgcc ttatccaaag atccaaacga aaagagagac
     4741 cacatggtct tgttagaatt tgttactgct gctggtatta cccatggtat ggatgaatta
     4801 tataaataag gcgcgccact tctaaataag cgaatttctt atgatttatg atttttatta
     4861 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     4921 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     4981 agtatgaggt cgctcttatt gaccacacct ctaccggcag atccgctagg gataacaggg
     5041 taatatagat ctgtttagct tgcctcgtcc ccgccgggtc acccggccag cgacatggag
     5101 gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt
     5161 cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat acattttgat
     5221 ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc ggacctgcga gcagggaaac
     5281 gctcccctca cagacgcgta taacttcgta taatgtatgc tatacgaagt tattgaattg
     5341 tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt
     5401 ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca
     5461 taaacaacca tgacagtcaa cactaagacc tatagtgaga gagcagaaac tcatgcctca
     5521 ccagtagcac aacgattatt tcgattaatg gaactgaaga aaaccaattt atgtgcatca
     5581 attgatgttg ataccactaa ggaattcctt gaattaattg ataaattggg tccttatgta
     5641 tgcttaatca agactcatat tgatataatc aatgattttt cctatgaatc cactattgaa
     5701 ccattattag aactttcacg taaacatcaa tttatgattt ttgaagatag aaaatttgct
     5761 gatattggta ataccgtgaa gaaacaatat attggtggag tttataaaat tagtagttgg
     5821 gcagatatta ctaatgctca tggtgtcact gggaatggag tagttgaagg attaaaacag
     5881 ggagctaaag aaaccaccac caaccaagag ccaagagggt tattgatgtt agctgaatta
     5941 tcatcagtgg gatcattagc atatggagaa tattctcaaa aaactgttga aattgctaaa
     6001 tccgataagg aatttgttat tggatttatt gcccaacgtg atatgggtgg acaagaagaa
     6061 ggatttgatt ggcttattat gacacctgga gttggattag atgataaagg tgatggatta
     6121 ggacaacaat atagaactgt tgatgaagtt gttagcactg gaactgatat tatcattgtt
     6181 ggtagaggat tgtttggtaa aggaagagat ccagatattg aaggtaaaag gtatagagat
     6241 gctggttgga atgcttattt gaaaaagact ggccaattat aatcagtact gacaataaaa
     6301 agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat
     6361 tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg
     6421 cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta
     6481 tactgctgtc gattcgatac taacgccgcc atccagtgtc gaaaacgagc tcgaattcat
     6541 aacttcgtat aatgtatgct atacgaagtt atatcgatga tatcagatcc actagtggcc
     6601 tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag
     6661 gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6721 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6781 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6841 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6901 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6961 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7021 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7081 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7141 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7201 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7261 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7321 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7381 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7441 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7501 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7561 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7621 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7681 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7741 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7801 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7861 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7921 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7981 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8041 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8101 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8161 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8221 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     8281 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8341 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8401 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8461 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8521 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8581 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8641 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     8701 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     8761 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     8821 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     8881 gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg cggctacaat
     8941 taatacataa ccttatgtat catacacata cgatttaggt ga
//
