LOCUS       sequence-269548-        5908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of SS-GFP-HDEL in genome, use auxotrophic marker 
            URA3(Saccharomyces cerevisiae) Marker for yeast endoplasmic 
            reticulum (ER)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5908)
  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 5908)
  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: ZJOM144
FEATURES             Location/Qualifiers
     source          1..5908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     177..196
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     430..447
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    564..585
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(600..605,606..623,624..630)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    638..654
                     /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     643..665
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     662..678
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     662..678
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     697..717
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        699..717
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        734..1253
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     CDS             1410..2123
                     /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"
     primer_bind     complement(2380..2399)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2381..2399)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2409..2426)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2409..2425)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2418..2440)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2566..3021
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2653..2672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2863..2884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3152..3955)
                     /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(3956..4171)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4195..4214)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4314..4336
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4374..4392)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4460..4564
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4565..4633,4634..5425)
                     /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(4783..4802)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5596..5908,1..276)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
       61 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
      121 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
      181 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
      241 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
      301 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
      361 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
      421 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
      481 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
      541 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
      601 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
      661 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
      721 caaaagctgg agccctacgt tagtgtgagc gggatttaaa ctgtgaggac cttaatacat
      781 tcagacactt ctgcggtatc accctactta ttcccttcga gattatatct aggaacccat
      841 caggttggtg gaagattacc cgttctaaga cttttcagct tcctctattg atgttacacc
      901 tggacacccc ttttctggca tccagttttt aatcttcagt ggcatgtgag attctccgaa
      961 attaattaaa gcaatcacac aattctctcg gataccacct cggttgaaac tgacaggtgg
     1021 tttgttacgc atgctaatgc aaaggagcct atataccttt ggctcggctg ctgtaacagg
     1081 gaatataaag ggcagcataa tttaggagtt tagtgaactt gcaacattta ctattttccc
     1141 ttcttacgta aatatttttc tttttaattc taaatcaatc tttttcaatt ttttgtttgt
     1201 attcttttct tgcttaaatc tataactaca aaaaacacat acataaacta aaaatgtttt
     1261 tcaacagact aagcgctggc aagctgctgg taccactctc cgtggtcctg tacgcccttt
     1321 tcgtggtaat attaccttta cagaattctt tccactcctc caatgtttta gttagaggtg
     1381 ccgatgatgg ggatccacta gtcgccacca tgtctaaagg tgaagaatta ttcactggtg
     1441 ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt tctgtctccg
     1501 gtgaaggtga aggtgatgct acttacggta aattgacctt aaaatttatt tgtactactg
     1561 gtaaattgcc agttccatgg ccaaccttag tcactacttt aacttatggt gttcaatgtt
     1621 tttctagata cccagatcat atgaaacaac atgacttttt caagtctgcc atgccagaag
     1681 gttatgttca agaaagaact atttttttca aagatgacgg taactacaag accagagctg
     1741 aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt attgatttta
     1801 aagaagatgg taacatttta ggtcacaaat tggaatacaa ctataactct cacaatgttt
     1861 acatcatggc tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt agacacaaca
     1921 ttgaagatgg ttctgttcaa ttagctgacc attatcaaca aaatactcca attggtgatg
     1981 gtccagtctt gttaccagac aaccattact tatccactca atctgcctta tccaaagatc
     2041 caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct ggtattaccc
     2101 atggtattga tgaattgtac aaaggagccg gaacaagtga catgaagcac gacgaattgt
     2161 agtcttttat atgaaaagaa atgaagcgtt aacatctaaa atcgaaatta tagggactca
     2221 ttagccatct attgttccag tataagaatg tttacataaa aaattcgaaa aaaaaagata
     2281 aaataaggga aaaatatata tatattcatt aagaagtttt gtacacaacg gagccaagat
     2341 ttcaccagta tctcgagggg gggcccggta cccaattcgc cctatagtga gtcgtattac
     2401 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2461 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2521 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg
     2581 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2641 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2701 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2761 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2821 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2881 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2941 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3001 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3061 ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt tgtgagttta
     3121 gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat
     3181 atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg
     3241 caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca tcatccacgg
     3301 ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg ggtgtcataa
     3361 tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa
     3421 caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata
     3481 gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt
     3541 cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg
     3601 taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat
     3661 taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg gcggataatg
     3721 cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt
     3781 ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc ttggtggtac
     3841 gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta aatagcttgg
     3901 cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc gacatgattt
     3961 atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact gggcaatttc
     4021 atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc tccttccttc
     4081 gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa
     4141 aaaaagaata aaaaaaaaat gatgaattga aaaggtggta tggtgcactc tcagtacaat
     4201 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4261 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4321 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4381 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4441 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4501 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4561 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4621 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4681 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4741 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4801 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4861 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4921 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4981 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5041 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5101 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5161 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5221 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5281 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5341 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5401 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5461 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5521 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5581 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5641 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5701 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5761 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5821 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5881 atagttaccg gataaggcgc agcggtcg
//
