LOCUS       sequence-210825-        6843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN/ARS plasmid expression of mitochondrial Su9-TagBFP marked with 
            HIS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6843)
  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 6843)
  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: pJW1664
FEATURES             Location/Qualifiers
     source          1..6843
                     /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        315..502
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             503..1165
                     /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"
     rep_origin      complement(1426..1881)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1563..1584)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1775..1794
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2011..2033
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2025..2042
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2026..2042
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2049..2068
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2049..2067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2098..2345)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2311..2329
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2352..3053)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     promoter        complement(3299..3942)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(3382..3405)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     misc_feature    3949..4056
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3973..3989
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3973..3989
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4023..4039)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4024..4040)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4069..4087)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4108..4124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4108..4124)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4121..4143)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4132..4148
                     /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(4156..4162,4163..4180,4181..4186))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4339..4356)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4510..5098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5269..6060,6061..6129))
                     /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     5892..5911
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6130..6234)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6271..6774
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6816..6834
                     /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 accataattc cgttttaaga gcttggtgag cgctaggagt cactgccagg tatcgtttga
      241 acacggcatt agtcagggaa gtcataacac agtcctttcc cgcaattttc tttttctatt
      301 actcttggcc tcctctagta cactctatat ttttttatgc ctcggtaatg attttcattt
      361 ttttttttcc acctagcgga tgactctttt tttttcttag cgattggcat tatcacataa
      421 tgaattatac attatataaa gtaatgtgat ttcttcgaag aatatactaa aaaatgagca
      481 ggcaagataa acgaaggcaa agatgacaga gcagaaagcc ctagtaaagc gtattacaaa
      541 tgaaaccaag attcagattg cgatctcttt aaagggtggt cccctagcga tagagcactc
      601 gatcttccca gaaaaagagg cagaagcagt agcagaacag gccacacaat cgcaagtgat
      661 taacgtccac acaggtatag ggtttctgga ccatatgata catgctctgg ccaagcattc
      721 cggctggtcg ctaatcgttg agtgcattgg tgacttacac atagacgacc atcacaccac
      781 tgaagactgc gggattgctc tcggtcaagc ttttaaagag gccctagggg ccgtgcgtgg
      841 agtaaaaagg tttggatcag gatttgcgcc tttggatgag gcactttcca gagcggtggt
      901 agatctttcg aacaggccgt acgcagttgt cgaacttggt ttgcaaaggg agaaagtagg
      961 agatctctct tgcgagatga tcccgcattt tcttgaaagc tttgcagagg ctagcagaat
     1021 taccctccac gttgattgtc tgcgaggcaa gaatgatcat caccgtagtg agagtgcgtt
     1081 caaggctctt gcggttgcca taagagaagc cacctcgccc aatggtacca acgatgttcc
     1141 ctccaccaaa ggtgttctta tgtagtgaca ccgattattt aaagctgcag catacgatat
     1201 atatacatgt gtatatatgt atacctatga atgtcagtaa gtatgtatac gaacagtatg
     1261 atactgaaga tgacaaggta atgcatcatt ctatacgtgt cattctgaac gaggcgcgct
     1321 ttcctttttt ctttttgctt tttctttttt tttctcttga actcgacgga tcatatgcgg
     1381 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaaacgtta
     1441 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1501 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1561 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1621 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     1681 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     1741 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     1801 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     1861 atgcgccgct acagggcgcg tcgcgccatt cgccattcag gctgcgcaac tgttgggaag
     1921 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaagggggga tgtgctgcaa
     1981 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     2041 gtgaattgta atacgactca ctatagggcg aattggagct ccaccgcggt ggcggccgca
     2101 aattaaagcc ttcgagcgtc ccaaaacctt ctcaagcaag gttttcagta taatgttaca
     2161 tgcgtacacg cgtctgtaca gaaaaaaaag aaaaatttga aatataaata acgttcttaa
     2221 tactaacata actataaaaa aataaatagg gacctagact tcaggttgtc taactccttc
     2281 cttttcggtt agagcggatg tggggggagg gcgtgaatgt aagcgtgaca taactaatta
     2341 catgactcga gttaattaag cttgtgcccc agtttgctag ggaggtcgca gtatctggcc
     2401 actgccacct cgtgctgctc gacgtaggtc tcgttgttgg cctccttgat tctttccagt
     2461 ctgtagtcca catagtagac gccaggcatc ttgaggttct tagcgggttt cttggatcta
     2521 tatgtggtct tgatgtttgc gatcagatgg ctcccgccca cgagcttcag ggccatgtcg
     2581 tttctgcctt ccaggccgcc gtcagcgggg tacagcgtct cggtgaaggc ctcccagccg
     2641 agtgttttct tctgcatcac agggccgttg gatgtgaagt tcacccctct gatcttgacg
     2701 ttgtagatga ggcagccgtc ctggaggctg gtgtcctggg tagcggtcag cacgcccccg
     2761 tcttcgtatg tggtgactct ctcccatgtg aagccctcag ggaaggactg cttgaagaag
     2821 tcggggatgc cctgggtgtg gttgatgaag gtcttgctgc cgtagaggaa gctagtagcc
     2881 aggatgtcga aggcgaaggg gagagggccg ccctcgacca ccttgattct catggtctgg
     2941 gtgccctcgt agggcttgcc ttcgccctcg gatgtgcact tgaagtgatg gttgtccacg
     3001 gtgccctcca tgtacagctt catgtgcatg ttctccttaa tcagctcgct catagatctg
     3061 gtacccgggg atccggaaga gtaggcgcgc ttctggaaag cctggcgggt ggtggcgttg
     3121 acgatggagg tcatctgggt gcgcttgagg gtctggaggg gggagccagt ctggatggtg
     3181 cgcttgctga cctgagcaac gcggacagca gggcgggcaa ccttggcgga agcagccatc
     3241 cgggaggcca ggcgagaggc gaggacacga gtggaggcca tactagttct agaatccgtc
     3301 gaaactaagt tctggtgttt taaaactaaa aaaaagacta actataaaag tagaatttaa
     3361 gaagtttaag aaatagattt acagaattac aatcaatacc taccgtcttt atatacttat
     3421 tagtcaagta ggggaataat ttcagggaac tggtttcaac cttttttttc agctttttcc
     3481 aaatcagaga gagcagaagg taatagaagg tgtaagaaaa tgagatagat acatgcgtgg
     3541 gtcaattgcc ttgtgtcatc atttactcca ggcaggttgc atcactccat tgaggttgtg
     3601 cccgtttttt gcctgtttgt gcccctgttc tctgtagttg cgctaagaga atggacctat
     3661 gaactgatgg ttggtgaaga aaacaatatt ttggtgctgg gattcttttt ttttctggat
     3721 gccagcttaa aaagcgggct ccattatatt tagtggatgc caggaataaa ctgttcaccc
     3781 agacacctac gatgttatat attctgtgta acccgccccc tattttgggc atgtacgggt
     3841 tacagcagaa ttaaaaggct aattttttga ctaaataaag ttaggaaaat cactactatt
     3901 aattatttac gtattctttg aaatggcgag tattgataat gataaactga gctccaccgc
     3961 ggtggcggcc gctctagaac tagtggatcc cccgggctgc aggaattcga tatcaagctt
     4021 atcgataccg tcgacctcga gggggggccc ggtacccagc ttttgttccc tttagtgagg
     4081 gttaattccg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4141 gctcacaatt ccacacaaca taggagccgg aagcataaag tgtaaagcct ggggtgccta
     4201 atgagtgagg taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4261 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4321 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4381 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4441 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4501 gctggcgttt ttccataggc tcggcccccc tgacgagcat cacaaaaatc gacgctcaag
     4561 tcagaggtgg cgaaacccga caggactata aagataccag gcgttccccc ctggaagctc
     4621 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4681 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     4741 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4801 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4861 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4921 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     4981 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5041 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5101 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5161 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5221 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5281 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5341 gcccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5401 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5461 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5521 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5581 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5641 ccaacgatca aggcgagtta catgatcccc catgttgtga aaaaaagcgg ttagctcctt
     5701 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5761 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5821 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5881 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5941 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6001 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6061 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6121 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6181 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6241 tccccgaaaa gtgccacctg ggtccttttc atcacgtgct ataaaaataa ttataattta
     6301 aattttttaa tataaatata taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa
     6361 aatagttttt gttttccgaa gatgtaaaag actctagggg gatcgccaac aaatactacc
     6421 ttttatcttg ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt
     6481 agaagaccac acacgaaaat cctgtgattt tacattttac ttatcgttaa tcgaatgtat
     6541 atctatttaa tctgcttttc ttgtctaata aatatatatg taaagtacgc tttttgttga
     6601 aattttttaa acctttgttt attttttttt cttcattccg taactcttct accttcttta
     6661 tttactttct aaaatccaaa tacaaaacat aaaaataaat aaacacagag taaattccca
     6721 aattattcca tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc gatccgtcct
     6781 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6841 gtc
//
