LOCUS       sequence-210836-        7374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN/ARS plasmid for GAL-inducible SWP1-V5-TEV-EGFP expression marked
            with URA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7374)
  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 7374)
  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: pJW1674
FEATURES             Location/Qualifiers
     source          1..7374
                     /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(3917..4358)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(3923..3946)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    4241..4358
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(4554..4570)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4555..4571)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4600..4618)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4639..4655)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4639..4655)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4652..4674)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4663..4679
                     /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(4687..4693,4694..4711,4712..4717))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4870..4887)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5041..5629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5800..6591,6592..6660))
                     /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     6423..6442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6661..6765)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6802..7305
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7347..7365
                     /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 cttaccgatt
     3061 gtttgaccaa aagatctcag gaacttggta gatgtcagta agcctggagc gcccaagtac
     3121 aaagaggaaa atagagtttc aaaaatactt gtacccaagt agtacctagc gaagataact
     3181 tcaaacccaa caatcgtggc gatgaatcct aggaaatata cagcagtgac accggtcggg
     3241 atgttattga atgcggcggc acaagaattc aaccaggtga ctattaaaga taaaatggta
     3301 atgaagataa ttaaaacaaa aatcacagca attggtttgg caactctctt tggctcggca
     3361 tgaaatatat gatggatttc tggcttaatg ccgaatttgt caactaaaga tgaatctgag
     3421 tgatctacat cgaagttcag gttgagttgc agaatttccc tgaacaagtt ttcctttggt
     3481 ttggcggtag atgacgctag aatcaaagtg gcctttattg gttccggtga tctggaagcc
     3541 tcttgcaaca aagcagcatc cagcttggct aaatctatcc tgtatttata catggacaac
     3601 ttgccattgt ccttaatttc gggttcaaaa gccatttcta ggttcttatt aggtaagcct
     3661 atcaataagg tgttctggaa gggtttgttg gtagactcaa tggcgaaatt tacttgaatc
     3721 acctcgttag ggtcttcaac tgtgattgta gtcggaacag tttcatcgat tcctattctg
     3781 ggttcaactt taccgatcat taccctagac tttcctgcgg tggaggggaa tgatacatga
     3841 acatcttgtg ccacgtaagc gaggacgaac gatatgcacg acaccaaggc cgcaagtgtt
     3901 ttgaagaatt gcatttctcc ttgacgttaa agtatagagg tatattaaca attttttgtt
     3961 gatactttta tgacatttga ataagaagta atacaaactg aaaatgttga aagtattagt
     4021 taaagtggtt atgcagcttt tccatttata tatctgttaa tagatcaaaa atcatcgctt
     4081 cgctgattaa ttaccccaga aacaaggcta aaaaactaat cgcattatca tcctatggtt
     4141 gttaatttga ttcgttaatt tgaaggtttg tggggccagg ttactgccaa tttttcctct
     4201 tcataaccat aaaagctagt attgtagaat ctttattgtt cggagcagtg cggcgcgagg
     4261 cacatctgcg tttcaggaac gcgaccggtg aagacgagga cgcacggagg agagtcttcc
     4321 gtcggagggc tgtcgcccgc tcggcggctt ctaatccgta cttcaatata gcaatgagca
     4381 gttaagcgta ttactgaaag ttccaaagag aaggtttttt taggctaaga taatggggct
     4441 ctttacattt ccacaacata taagtaagat tagatatgga tatgtatatg gtggtaatgc
     4501 catgtaatat gattattaaa cttctttgcg tccatccaag atatcaagct tatcgatacc
     4561 gtcgacctcg agggggggcc cggtacccag cttttgttcc ctttagtgag ggttaattcc
     4621 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4681 tccacacaac ataggagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4741 gtaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4801 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4861 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4921 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4981 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5041 tttccatagg ctcggccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5101 gcgaaacccg acaggactat aaagatacca ggcgttcccc cctggaagct ccctcgtgcg
     5161 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5221 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5281 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5341 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5401 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5461 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5521 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5581 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5641 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5701 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5761 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5821 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tgcccgtcgt
     5881 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5941 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6001 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6061 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6121 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6181 aaggcgagtt acatgatccc ccatgttgtg aaaaaaagcg gttagctcct tcggtcctcc
     6241 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6301 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6361 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6421 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6481 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6541 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6601 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6661 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6721 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6781 agtgccacct gggtcctttt catcacgtgc tataaaaata attataattt aaatttttta
     6841 atataaatat ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt
     6901 tgttttccga agatgtaaaa gactctaggg ggatcgccaa caaatactac cttttatctt
     6961 gctcttcctg ctctcaggta ttaatgccga attgtttcat cttgtctgtg tagaagacca
     7021 cacacgaaaa tcctgtgatt ttacatttta cttatcgtta atcgaatgta tatctattta
     7081 atctgctttt cttgtctaat aaatatatat gtaaagtacg ctttttgttg aaatttttta
     7141 aacctttgtt tatttttttt tcttcattcc gtaactcttc taccttcttt atttactttc
     7201 taaaatccaa atacaaaaca taaaaataaa taaacacaga gtaaattccc aaattattcc
     7261 atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag cgatccgtcc taagaaacca
     7321 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
