LOCUS       sequence-352610-        6860 bp    DNA     circular SYN 14-APR-2025
DEFINITION  mCherry and the 1-10th beta-strands of GFP (GFP1-10) fused to a 
            mitochondria signal sequence for mitochondrial targeting in yeast 
            plasmid with HIS marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6860)
  AUTHORS   Cortes Sanchon A, Santhosh Kumar H, Mantovani M, Osinnii I, Mateos 
            JM, Kaech A, Shcherbakov D, Akbergenov R, Bottger EC
  TITLE     ER-misfolded proteins become sequestered with mitochondria and 
            impair mitochondrial function.
  JOURNAL   Commun Biol. 2021 Dec 2;4(1):1350. doi: 10.1038/s42003-021-02873-w.
  PUBMED    34857875
REFERENCE   2  (bases 1 to 6860)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2021-12-2"; volume: "4(1):1350. doi"; pages: " 
            10.1038/s42003-021-02873-w"
COMMENT     Sequence Label: p403 – TDH3-MTS-mCherry-GFP1-10-His
FEATURES             Location/Qualifiers
     source          1..6860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..65
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(103..121)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        189..293
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(294..362,363..1154)
                     /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(512..531)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1325..1913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1814..1833
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2067..2084
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2201..2222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2237..2242,2243..2260,2261..2267)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2275..2291
                     /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     2280..2302
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2299..2315
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2299..2315
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2334..2354
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2336..2354
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2379..3022
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2916..2939
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     regulatory      3268..3277
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      4654..4901
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4670..4688)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4919..4938)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4920..4938)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4948..4965)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4948..4964)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4957..4979)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5105..5560
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5192..5211)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5402..5423
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5787..6449)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(6450..6637)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(6784..6803)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
       61 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
      121 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
      181 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      241 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      301 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      361 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      421 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      481 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      541 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
      601 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
      661 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      721 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      781 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
      841 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      901 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      961 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1021 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1081 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1141 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1201 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1261 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1321 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1381 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1441 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1501 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1561 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1621 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1681 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1741 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1801 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1861 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1921 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     1981 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2041 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2101 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2161 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2221 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2281 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     2341 accctcacta aagggaacaa aagctggagc tcagtttatc attatcaata ctcgccattt
     2401 caaagaatac gtaaataatt aatagtagtg attttcctaa ctttatttag tcaaaaaatt
     2461 agccttttaa ttctgctgta acccgtacat gcccaaaata gggggcgggt tacacagaat
     2521 atataacatc gtaggtgtct gggtgaacag tttattcctg gcatccacta aatataatgg
     2581 agcccgcttt ttaagctggc atccagaaaa aaaaagaatc ccagcaccaa aatattgttt
     2641 tcttcaccaa ccatcagttc ataggtccat tctcttagcg caactacaga gaacaggggc
     2701 acaaacaggc aaaaaacggg cacaacctca atggagtgat gcaacctgcc tggagtaaat
     2761 gatgacacaa ggcaattgac ccacgcatgt atctatctca ttttcttaca ccttctatta
     2821 ccttctgctc tctctgattt ggaaaaagct gaaaaaaaag gttgaaacca gttccctgaa
     2881 attattcccc tacttgacta ataagtatat aaagacggta ggtattgatt gtaattctgt
     2941 aaatctattt cttaaacttc ttaaattcta cttttatagt tagtcttttt tttagtttta
     3001 aaacaccaga acttagtttc gacggattct agaactagta tggcctccac tcgtgtcctc
     3061 gcctctcgcc tggcctcccg gatggctgct tccgccaagg ttgcccgccc tgctgtccgc
     3121 gttgctcagg tcagcaagcg caccatccag actggctccc ccctccagac cctcaagcgc
     3181 acccagatga cctccatcgt caacgccacc acccgccagg ctttccagaa gcgcgcctac
     3241 tcttccggat ccggtgacgg tccggtcgcc accatggttt ctaaaggtga agaagataat
     3301 atggctatta ttaaagaatt tatgagattt aaagttcata tggaaggttc tgttaatggt
     3361 catgaatttg aaattgaagg tgaaggtgaa ggtagaccat atgaaggtac tcaaactgct
     3421 aaattgaaag ttactaaagg tggtccattg ccatttgctt gggatatttt gtctccacaa
     3481 tttatgtatg gttctaaagc ttatgttaaa catccagctg atattccaga ttatttgaaa
     3541 ttgtcttttc cagaaggttt taaatgggaa agagttatga attttgaaga tggtggtgtt
     3601 gttactgtta ctcaagattc ttctttgcaa gatggtgaat ttatttataa agttaaattg
     3661 agaggtacta attttccatc tgatggtcca gttatgcaaa aaaaaactat gggttgggaa
     3721 gcttcttctg aaagaatgta tccagaagat ggtgctttga aaggtgaaat taaacaaaga
     3781 ttgaaattga aagatggtgg tcattatgat gctgaagtta aaactactta taaagctaaa
     3841 aaaccagttc aattgccagg tgcttataat gttaatatta aattggatat tacttctcat
     3901 aatgaagatt atactattgt tgaacaatat gaaagagctg aaggtagaca ttctactggt
     3961 ggtatggatg aattgtataa agaattcggt ggtggttctg gcggatctaa aggtgaagaa
     4021 ttgtttactg gtgttgttcc aattttggtt gaattggatg gtgatgttaa tggtcataaa
     4081 ttttctgtta gaggtgaagg tgaaggtgat gctactattg gtaaattgac tttgaaattt
     4141 atttgtacta ctggtaaatt gccagttcca tggccaactt tggttactac tttgacttat
     4201 ggtgttcaat gtttttctag atatccagat catatgaaac aacatgattt ttttaaatct
     4261 gctatgccag aaggttatgt tcaagaaaga actatttctt ttaaagatga tggtaaatat
     4321 aaaactagag ctgttgttaa atttgaaggt gatactttgg ttaatagaat tgaattgaaa
     4381 ggtactgatt ttaaagaaga tggtaatatt ttgggtcata aattggaata taattttaat
     4441 tctcataatg tttatattac tgctgataaa caaaaaaatg gtattaaagc taattttact
     4501 gttagacata atgttgaaga tggttctgtt caattggctg atcattatca acaaaatact
     4561 ccaattggtg atggtccagt tttgttgcca gataatcatt atttgtctac tcaaactgtt
     4621 ttgtctaaag atccaaatga aaaataactc gagtcatgta attagttatg tcacgcttac
     4681 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     4741 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     4801 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     4861 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccggtac ccaattcgcc
     4921 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     4981 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     5041 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     5101 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5161 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5221 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5281 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5341 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5401 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5461 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5521 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     5581 ttacgcatct gtgcggtatt tcacaccgca tagggtaata actgatataa ttaaattgaa
     5641 gctctaattt gtgagtttag tatacatgca ttaccttgtc atcttcagta tcatactgtt
     5701 cgtatacata cttactgaca ttcataggta tacatatata cacatgtata tatatcgtat
     5761 gctgcagctt taaataatcg gtgtcactac ataagaacac ctttggtgga gggaacatcg
     5821 ttggtaccat tgggcgaggt ggcttctctt atggcaaccg caagagcctt gaacgcactc
     5881 tcactacggt gatgatcatt cttgcctcgc agacaatcaa cgtggagggt aattctgcta
     5941 gcctctgcaa agctttcaag aaaatgcggg atcatctcgc aagagagatc tcctactttc
     6001 tccctttgca aaccaagttc gacaactgcg tacggcctgt tcgaaagatc taccaccgct
     6061 ctggaaagtg cctcatccaa aggcgcaaat cctgatccaa acctttttac tccacgcacg
     6121 gcccctaggg cctctttaaa agcttgaccg agagcaatcc cgcagtcttc agtggtgtga
     6181 tggtcgtcta tgtgtaagtc accaatgcac tcaacgatta gcgaccagcc ggaatgcttg
     6241 gccagagcat gtatcatatg gtccagaaac cctatacctg tgtggacgtt aatcacttgc
     6301 gattgtgtgg cctgttctgc tactgcttct gcctcttttt ctgggaagat cgagtgctct
     6361 atcgctaggg gaccaccctt taaagagatc gcaatctgaa tcttggtttc atttgtaata
     6421 cgctttacta gggctttctg ctctgtcatc tttgccttcg tttatcttgc ctgctcattt
     6481 tttagtatat tcttcgaaga aatcacatta ctttatataa tgtataattc attatgtgat
     6541 aatgccaatc gctaagaaaa aaaaagagtc atccgctagg tggaaaaaaa aaaatgaaaa
     6601 tcattaccga ggcataaaaa aatatagagt gtactagagg aggccaagag taatagaaaa
     6661 agaaaattgc gggaaaggac tgtgttatga cttccctgac taatgccgtg ttcaaacgat
     6721 acctggcagt gactcctagc gctcaccaag ctcttaaaac gggaatttat ggtgcactct
     6781 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     6841 tgacgcgccc tgacgggctt
//
