LOCUS       sequence-256863-        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"
     primer_bind     complement(12..29)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(263..282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(942..1733,1734..1802))
                     /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     1565..1584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1803..1907)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1975..1993
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1995..2096
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             2097..2771
                     /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(2890..2912)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3012..3031
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3100..3619
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     regulatory      3770..3779
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3776..4483
                     /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(3921..3939)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4189..4208
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      4509..4756
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4525..4543)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4775..4791)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4775..4791)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4788..4810)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4799..4815
                     /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(4823..4829,4830..4847,4848..4853))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4868..4889
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     1981 cgtatcacga ggccaattcg gtcgaaaaaa gaaaaggaga gggccaagag ggagggcatt
     2041 ggtgactatt gagcacgtga gtatacgtga ttaagcacac aaaggcagct tggagtatgt
     2101 ctgttattaa tttcacaggt agttctggtc cattggtgaa agtttgcggc ttgcagagca
     2161 cagaggccgc agaatgtgca ctagattccg atgctgactt gctgggtatt atatgtgtgc
     2221 ccaatagaaa gagaacaatt gacccggtta ttgcaaggaa aatttcaagt cttgtaaaag
     2281 catataaaaa tagttcaggc actccgaaat acttggttgg cgtgtttcgt aatcaaccta
     2341 aggaggatgt tttggctctg gtcaatgatt acggcattga tatcgtccaa ctgcatggag
     2401 atgagtcgtg gcaagaatac caagagttcc tcggtttgcc agttattaaa agactcgtat
     2461 ttccaaaaga ctgcaacata ctactcagtg cagcttcaca gaaacctcat tcgtttattc
     2521 ccttgtttga ttcagaagca ggtgggacag gtgaactttt ggattggaac tcgatttctg
     2581 actgggttgg aaggcaagag agccccgaga gcttacattt tatgttagct ggtggactga
     2641 cgccagaaaa tgttggtgat gcgcttagat taaatggcgt tattggtgtt gatgtaagcg
     2701 gaggtgtgga gacaaatggt gtaaaagact ctaacaaaat agcaaatttc gtcaaaaatg
     2761 ctaagaaata ggttattact gagtagtatt tatttaagta ttgtttgtgc acttgcctgc
     2821 aagccttttg aaaagcaagc ataaaagatc gccctttcgt ctcgcgcgtt tcggtgatga
     2881 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     2941 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     3001 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     3061 accgcacaga tgcgtaagga gaaaataccg catcaggcgc ctacgttagt gtgagcggga
     3121 tttaaactgt gaggacctta atacattcag acacttctgc ggtatcaccc tacttattcc
     3181 cttcgagatt atatctagga acccatcagg ttggtggaag attacccgtt ctaagacttt
     3241 tcagcttcct ctattgatgt tacacctgga cacccctttt ctggcatcca gtttttaatc
     3301 ttcagtggca tgtgagattc tccgaaatta attaaagcaa tcacacaatt ctctcggata
     3361 ccacctcggt tgaaactgac aggtggtttg ttacgcatgc taatgcaaag gagcctatat
     3421 acctttggct cggctgctgt aacagggaat ataaagggca gcataattta ggagtttagt
     3481 gaacttgcaa catttactat tttcccttct tacgtaaata tttttctttt taattctaaa
     3541 tcaatctttt tcaatttttt gtttgtattc ttttcttgct taaatctata actacaaaaa
     3601 acacatacat aaactaaaaa tgtttttcaa cagactaagc gctggcaagc tgctggtacc
     3661 actctccgtg gtcctgtacg cccttttcgt ggtaatatta cctttacaga attctttcca
     3721 ctcctccaat gttttagtta gaggtgccga tgatggggat ccactagtcg ccaccatggt
     3781 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     3841 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     3901 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     3961 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     4021 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     4081 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     4141 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     4201 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     4261 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     4321 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     4381 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     4441 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagcacgacg aattgtaatc
     4501 tagagtcatc atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc
     4561 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat
     4621 agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag
     4681 acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc
     4741 gaaggcttta atttgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4801 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4861 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4921 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4981 tgcgtattgg gcgc
//
