LOCUS       sequence-210834-        7848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN/ARS plasmid for GAL-inducible PST1-V5-TEV-EGFP expression marked
            with URA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7848)
  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 7848)
  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: pJW1672
FEATURES             Location/Qualifiers
     source          1..7848
                     /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(4391..4832)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(4397..4420)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    4715..4832
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(5028..5044)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5029..5045)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5074..5092)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5113..5129)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5113..5129)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5126..5148)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5137..5153
                     /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(5161..5167,5168..5185,5186..5191))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5344..5361)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5515..6103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6274..7065,7066..7134))
                     /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     6897..6916
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7135..7239)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7276..7779
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7821..7839
                     /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 cttacccatt
     3061 atagacatga tgattgccgt caaaagaccg accaaagggg ttgtttggcc aattggtgcc
     3121 atgatagcgg cattgccttt ggaactctta gagctagagc tggaagcgcc agaactagaa
     3181 acactagagg cagaagcagc agccttgata tcgccagaag tgaacttctt ttcctcggcc
     3241 ttagaagatg acttggaaac cttggcagta gtttggcttg attgagattt ggaagtggac
     3301 gatagtttaa cagatgtgga tgatgcacca tttttgcaga caaaagattc acccttgata
     3361 ccccctttct tttgcaaagc tttcaaagca ttacaggaga aattgcttga ctttgattcg
     3421 acatctgcgc cacccttgac agacttcaaa gagtctaggt tcaaggaggt gaagttacca
     3481 acaacttcca aagtaccgcc aatggttgtt aggttgtcga gaccaccaat tttttgtaaa
     3541 ccagtgttgt tagcaacgac aagagcacca cctatggttg acaaattaga gaacgacaag
     3601 ttcttcaagt agtcattgga aacgatactg aaggtttggc caatggtgtt tagcttagtg
     3661 aaattcaaac ttgagatgga gttgttgatg aaacccaatg aagagttaat cttttgcaag
     3721 ttagcgaagg aaacagagtg gacatcggtc aaactgatat tgtttgccca aaccaagtca
     3781 tcgaaggtga ttttagtctg gttaccgttg aacgaaaatt gtaaagaatc gctgactgtt
     3841 tcaactggag atttgatcga ggttaatttc ttattgttat tgacgttgaa cacgttaact
     3901 tttttcaagg ctgagaatcc atcgacagat tgtaacgaag tgtcggaaat ataaatgttg
     3961 ttagcagatt tgatatttga agtaaaactg gagatggcgg gtagagtaat cagttttata
     4021 ctatcaacgc tctgtaaaga cccaaatgaa gcagaagtca agattgtcaa actctgtagg
     4081 ttcaaagaat ctgtgatgga ctccaaggaa tcagcagcga atgaggttag atttgtagcg
     4141 ttaaatatag ttagagaccc gttgatttct ttaacattag ccaaagcacc agtcttcaaa
     4201 ccaccaccaa tagttaagtt cccgactaac gtatcacagc ggctatattt atctaagtca
     4261 ctctgagctg tggccgtggc atgtgagctt atggtacagg aagagggtat gctggaagaa
     4321 gaggaagtag cagccaaagc tgacgttatt aagagcgcag ttgaagcgat aagtgaatgt
     4381 aattgcattt ctccttgacg ttaaagtata gaggtatatt aacaattttt tgttgatact
     4441 tttatgacat ttgaataaga agtaatacaa actgaaaatg ttgaaagtat tagttaaagt
     4501 ggttatgcag cttttccatt tatatatctg ttaatagatc aaaaatcatc gcttcgctga
     4561 ttaattaccc cagaaacaag gctaaaaaac taatcgcatt atcatcctat ggttgttaat
     4621 ttgattcgtt aatttgaagg tttgtggggc caggttactg ccaatttttc ctcttcataa
     4681 ccataaaagc tagtattgta gaatctttat tgttcggagc agtgcggcgc gaggcacatc
     4741 tgcgtttcag gaacgcgacc ggtgaagacg aggacgcacg gaggagagtc ttccgtcgga
     4801 gggctgtcgc ccgctcggcg gcttctaatc cgtacttcaa tatagcaatg agcagttaag
     4861 cgtattactg aaagttccaa agagaaggtt tttttaggct aagataatgg ggctctttac
     4921 atttccacaa catataagta agattagata tggatatgta tatggtggta atgccatgta
     4981 atatgattat taaacttctt tgcgtccatc caagatatca agcttatcga taccgtcgac
     5041 ctcgaggggg ggcccggtac ccagcttttg ttccctttag tgagggttaa ttccgagctt
     5101 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5161 caacatagga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgaggtaact
     5221 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5281 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5341 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5401 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5461 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5521 taggctcggc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5581 cccgacagga ctataaagat accaggcgtt cccccctgga agctccctcg tgcgctctcc
     5641 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5701 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5761 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5821 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5881 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5941 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6001 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6061 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6121 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6181 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6241 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6301 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactgcccg tcgtgtagat
     6361 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6421 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6481 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6541 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6601 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6661 agttacatga tcccccatgt tgtgaaaaaa agcggttagc tccttcggtc ctccgatcgt
     6721 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6781 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6841 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6901 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6961 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7021 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7081 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7141 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7201 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7261 acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa
     7321 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt
     7381 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt
     7441 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg
     7501 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc
     7561 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt
     7621 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat
     7681 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt
     7741 aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcctaagaa accattatta
     7801 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
