LOCUS       sequence-292102-        4994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of SS-mCherry-HDEL in genome, use auxotrophic marker 
            TRP1(Saccharomyces cerevisiae). Marker for yeast endoplasmic 
            reticulum (ER)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4994)
  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 4994)
  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: ZJOM90
FEATURES             Location/Qualifiers
     source          1..4994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..521
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     regulatory      672..681
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             678..1385
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(823..841)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1091..1110
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1411..1658
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1427..1445)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1677..1693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1677..1693)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1690..1712)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1701..1717
                     /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(1725..1731,1732..1749,1750..1755))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1770..1791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1908..1925)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2079..2667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2159..2178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2838..3629,3630..3698))
                     /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     3461..3480
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3699..3803)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3871..3889
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3891..3992
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             3993..4667
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     primer_bind     complement(4786..4808)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4908..4927
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcctacgtta gtgtgagcgg gatttaaact gtgaggacct taatacattc agacacttct
       61 gcggtatcac cctacttatt cccttcgaga ttatatctag gaacccatca ggttggtgga
      121 agattacccg ttctaagact tttcagcttc ctctattgat gttacacctg gacacccctt
      181 ttctggcatc cagtttttaa tcttcagtgg catgtgagat tctccgaaat taattaaagc
      241 aatcacacaa ttctctcgga taccacctcg gttgaaactg acaggtggtt tgttacgcat
      301 gctaatgcaa aggagcctat atacctttgg ctcggctgct gtaacaggga atataaaggg
      361 cagcataatt taggagttta gtgaacttgc aacatttact attttccctt cttacgtaaa
      421 tatttttctt tttaattcta aatcaatctt tttcaatttt ttgtttgtat tcttttcttg
      481 cttaaatcta taactacaaa aaacacatac ataaactaaa aatgtttttc aacagactaa
      541 gcgctggcaa gctgctggta ccactctccg tggtcctgta cgcccttttc gtggtaatat
      601 tacctttaca gaattctttc cactcctcca atgttttagt tagaggtgcc gatgatgggg
      661 atccactagt cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
      721 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
      781 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
      841 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
      901 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
      961 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     1021 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     1081 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     1141 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     1201 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     1261 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     1321 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     1381 acaagcacga cgaattgtaa tctagagtca tcatgtaatt agttatgtca cgcttacatt
     1441 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
     1501 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
     1561 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
     1621 ttgagaaggt tttgggacgc tcgaaggctt taatttgcaa gcttggcgta atcatggtca
     1681 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1741 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1801 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1861 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1921 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1981 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2041 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2101 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2161 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2221 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2281 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2341 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2401 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2461 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2521 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2581 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2641 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2701 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2761 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2821 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2881 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2941 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3001 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3061 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3121 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3181 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3241 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3301 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3361 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3421 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3481 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3541 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3601 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3661 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3721 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3781 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     3841 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc aattcggtcg
     3901 aaaaaagaaa aggagagggc caagagggag ggcattggtg actattgagc acgtgagtat
     3961 acgtgattaa gcacacaaag gcagcttgga gtatgtctgt tattaatttc acaggtagtt
     4021 ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga ggccgcagaa tgtgcactag
     4081 attccgatgc tgacttgctg ggtattatat gtgtgcccaa tagaaagaga acaattgacc
     4141 cggttattgc aaggaaaatt tcaagtcttg taaaagcata taaaaatagt tcaggcactc
     4201 cgaaatactt ggttggcgtg tttcgtaatc aacctaagga ggatgttttg gctctggtca
     4261 atgattacgg cattgatatc gtccaactgc atggagatga gtcgtggcaa gaataccaag
     4321 agttcctcgg tttgccagtt attaaaagac tcgtatttcc aaaagactgc aacatactac
     4381 tcagtgcagc ttcacagaaa cctcattcgt ttattccctt gtttgattca gaagcaggtg
     4441 ggacaggtga acttttggat tggaactcga tttctgactg ggttggaagg caagagagcc
     4501 ccgagagctt acattttatg ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc
     4561 ttagattaaa tggcgttatt ggtgttgatg taagcggagg tgtggagaca aatggtgtaa
     4621 aagactctaa caaaatagca aatttcgtca aaaatgctaa gaaataggtt attactgagt
     4681 agtatttatt taagtattgt ttgtgcactt gcctgcaagc cttttgaaaa gcaagcataa
     4741 aagatcgccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4801 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4861 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     4921 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     4981 ataccgcatc aggc
//
