LOCUS       sequence-210829-        8130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN/ARS plasmid for GAL-inducible ERG5-V5-TEV-EGFP expression marked
            with URA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8130)
  AUTHORS   Costa EA, Subramanian K, Nunnari J, Weissman JS
  TITLE     Defining the physiological role of SRP in protein-targeting 
            efficiency and specificity.
  JOURNAL   Science. 2018 Jan 18. pii: science.aar3607. doi: 
            10.1126/science.aar3607.
  PUBMED    29348368
REFERENCE   2  (bases 1 to 8130)
  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: "Science. 
            2018 Jan 18. pii: science.aar3607. doi: 10.1126/science.aar3607."
COMMENT     Sequence Label: pJW1667
FEATURES             Location/Qualifiers
     source          1..8130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        190..410
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             411..1214
                     /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"
     rep_origin      complement(1343..1798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1480..1501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1692..1711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1928..1950
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1942..1959
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1943..1959
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2017..2033
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2017..2033
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      complement(2061..2226)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     2147..2171
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2247..2963)
                     /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             complement(2250..2963)
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(2988..3008)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(3015..3056)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     promoter        complement(4673..5114)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(4679..4702)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    4997..5114
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(5310..5326)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5311..5327)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5356..5374)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5395..5411)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5395..5411)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5408..5430)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5419..5435
                     /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(5443..5449,5450..5467,5468..5473))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5626..5643)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5797..6385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6556..7347,7348..7416))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7179..7198
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7417..7521)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7558..8061
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8103..8121
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
      241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
      301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
      361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
      421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
      481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
      541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
      601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
      661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
      721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
      781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
      841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
      901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
      961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     1261 gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt
     1321 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta
     1381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     1441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     1501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     1561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     1621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     1681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     1741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
     1801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1861 ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca
     1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     1981 tagggcgaat tggagctcca ccgcggtggc ggccgctcta gaactagtgg atcccccggg
     2041 ctgcagggtg tggtcaataa gagcgacctc atactatacc tgagaaagca acctgaccta
     2101 caggaaagag ttactcaaga ataagaattt tcgttttaaa acctaagagt cactttaaaa
     2161 tttgtataca cttatttttt ttataactta tttaataata aaaatcataa atcataagaa
     2221 attcgcttat ttagaagtgg cgcgccttat ttgtacaatt catccatacc atgggtaata
     2281 ccagcagcag taacaaattc taacaagacc atgtggtctc tcttttcgtt tggatctttg
     2341 gataaggcag attgagtgga taagtaatgg ttgtctggta acaagactgg accatcacca
     2401 attggagtat tttgttgata atggtcagct aattgaacag aaccatcttc aatgttgtgt
     2461 ctaattttga agttaacttt gataccattc ttttgtttgt cagccatgat gtaaacattg
     2521 tgagagttat agttgtattc caatttgtga cctaaaatgt taccatcttc tttaaaatca
     2581 atacctttta attcgattct attaactaag gtatcacctt caaacttgac ttcagctctg
     2641 gtcttgtagt taccgtcatc tttgaaaaaa atagttcttt cttgaacata accttctggc
     2701 atggcagact tgaaaaagtc atgttgtttc atatgatctg ggtatctaga aaaacattga
     2761 acaccataag ttaaagtagt gactaaggtt ggccatggaa ctggcaattt accagtagta
     2821 caaataaatt ttaaggtcaa tttaccgtaa gtagcatcac cttcaccttc accggagaca
     2881 gaaaatttgt gaccattaac atcaccatct aattcaacca aaattgggac aacaccagtg
     2941 aataattctt cacctttaga catgttaatt aaaccagcac cgtcacctcc ttgaaaatac
     3001 aaattttctg aacccgtaga atcgagaccg aggagagggt tagggatagg cttaccttcg
     3061 aagacttctc cagtaattgg gtctctcttt ttgaaagtca gtaacaaatc atcttttggg
     3121 aaaattgtag cgaaaacctt gattttttca cttaatggag tcactgtatg atggaaatca
     3181 gtatatagtg caaatttacc caacaaagcg gcgaaggtaa tcatgacata tgtttgacct
     3241 aagcaaacgt gtggaccaca accaaaaacc aaccaattct tctttgcttc actagcctta
     3301 gagccttcta cccatctttc agggatgaac tcatcaggat tttcgtaaac ttcaggatca
     3361 tgtaaagctg ggtataaggt tggaattaac atagcgccct ttggtgcggt atagttaggg
     3421 gaaactggga aattcttctt aacaacatat ggaaccatca agacaggagg tctgtaacgc
     3481 aaagtttctt ttatgaccat attggtgtac ttcattttct caatcaaatc caagttcaat
     3541 tcggtagaca tgtcattgtt acgaacagcc aattgttctt ctctgatctt agctaagaca
     3601 tctggacggt cagcaacaat ttggaacaac caacaagcta aagaagaaga ggcatcttga
     3661 gaagcaaata agaaagtgaa aacagcttcg gagatttcct tgttagtaaa ctctctgtgg
     3721 tagattctag aatcatcatc gttactattc tttgcatcgt gcatcaactt acaccaagca
     3781 tccataacac aaactggctt accacctgca gcaatatgat ccttagccat ttgagcacag
     3841 ttttcgaaaa tcttcatggc catgtctgca gttttcttac cataccatgt tttagtgtaa
     3901 gggataataa ttgggaagtt gactaattcc aatgctgctg taaccaaata gtaatcatca
     3961 gcaatctttc tgacttgatc ttcggtaata tagttaccac agaaagagtt caatgataag
     4021 gcgcaaagaa tttctctcat ttcatggaaa aagacctggg gctcgtagtt attctcctta
     4081 gataaacgaa caaacttatc catgtactta tccatgattt gttccaatga aggtaagtat
     4141 tgagccaaag cttgtttagt gaaaagaccg tttaatgatt ttctgtaatc agtatgagct
     4201 ttaccgtcca aaaaaaccca attgcaaggt cttaagatct tcacagcaac atcgacaacg
     4261 caaggtttga cgaatttgga agattgcaag atctttcttg ccaagtctct agtagatgcg
     4321 ataacaacaa atttatggaa aatagaaaca catgaaagtg gaccggatgc ccacttagcc
     4381 ttatattctt caaactttgg atctaaggat tccaaaaatg gaccgatgat gggccagaac
     4441 ttaaactttg gacctgcgat ggaacctttc ttgatttgat atgcaacttg gtcccaaacc
     4501 aaaagaatac agattaaagt agcaaatatt ttcaaatatg acatagactt aagtgtatcc
     4561 aggatactga aggcagtagt gacgcctaca gagggctggt ctttagccaa ttggtgtagc
     4621 gtagaattat gagtggcatg ttgtattata ttttctgcga cagaactcat ttctccttga
     4681 cgttaaagta tagaggtata ttaacaattt tttgttgata cttttatgac atttgaataa
     4741 gaagtaatac aaactgaaaa tgttgaaagt attagttaaa gtggttatgc agcttttcca
     4801 tttatatatc tgttaataga tcaaaaatca tcgcttcgct gattaattac cccagaaaca
     4861 aggctaaaaa actaatcgca ttatcatcct atggttgtta atttgattcg ttaatttgaa
     4921 ggtttgtggg gccaggttac tgccaatttt tcctcttcat aaccataaaa gctagtattg
     4981 tagaatcttt attgttcgga gcagtgcggc gcgaggcaca tctgcgtttc aggaacgcga
     5041 ccggtgaaga cgaggacgca cggaggagag tcttccgtcg gagggctgtc gcccgctcgg
     5101 cggcttctaa tccgtacttc aatatagcaa tgagcagtta agcgtattac tgaaagttcc
     5161 aaagagaagg tttttttagg ctaagataat ggggctcttt acatttccac aacatataag
     5221 taagattaga tatggatatg tatatggtgg taatgccatg taatatgatt attaaacttc
     5281 tttgcgtcca tccaagatat caagcttatc gataccgtcg acctcgaggg ggggcccggt
     5341 acccagcttt tgttcccttt agtgagggtt aattccgagc ttggcgtaat catggtcata
     5401 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatag gagccggaag
     5461 cataaagtgt aaagcctggg gtgcctaatg agtgaggtaa ctcacattaa ttgcgttgcg
     5521 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5581 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5641 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5701 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5761 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcg gcccccctga
     5821 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5881 ataccaggcg ttcccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5941 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     6001 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6061 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6121 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6181 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6241 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6301 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6361 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6421 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6481 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6541 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6601 atttcgttca tccatagttg cctgactgcc cgtcgtgtag ataactacga tacgggaggg
     6661 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6721 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6781 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6841 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6901 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6961 gttgtgaaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7021 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7081 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7141 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7201 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7261 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7321 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7381 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7441 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7501 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt ccttttcatc
     7561 acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa attaaaaata
     7621 gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat gtaaaagact
     7681 ctagggggat cgccaacaaa tactaccttt tatcttgctc ttcctgctct caggtattaa
     7741 tgccgaattg tttcatcttg tctgtgtaga agaccacaca cgaaaatcct gtgattttac
     7801 attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg tctaataaat
     7861 atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt tttttttctt
     7921 cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac aaaacataaa
     7981 aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata cgaggcgcgt
     8041 gtaagttaca ggcaagcgat ccgtcctaag aaaccattat tatcatgaca ttaacctata
     8101 aaaataggcg tatcacgagg ccctttcgtc
//
