LOCUS       sequence-102449-        6782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS316-pTDH3-Su9-TagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6782)
  AUTHORS   Williams CC, Jan CH, Weissman JS
  TITLE     Targeting and plasticity of mitochondrial proteins revealed by 
            proximity-specific ribosome profiling.
  JOURNAL   Science. 2014 Nov 7;346(6210):748-51. doi: 10.1126/science.1257522.
  PUBMED    25378625
REFERENCE   2  (bases 1 to 6782)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2014-11-7- 7"; volume: "346"; issue: 
            "6210"; pages: "748-51"
COMMENT     Sequence Label: pJW1513
FEATURES             Location/Qualifiers
     source          1..6782
                     /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(1378..1833)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1515..1536)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1727..1746
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1959..1981
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1973..1990
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1974..1990
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2000..2019
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2000..2018
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2037..2284)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2250..2268
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2291..2992)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     promoter        complement(3238..3881)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(3321..3344)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     misc_feature    3888..3995
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3912..3928
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3912..3928
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3962..3978)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3963..3979)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4008..4026)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4047..4063)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4047..4063)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4060..4082)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4071..4087
                     /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(4095..4101,4102..4119,4120..4125))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4278..4295)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4449..5037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5208..5999,6000..6068))
                     /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     5831..5850
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6069..6173)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6210..6713
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6755..6773
                     /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 taaatgcatc attctatacg tgtcattctg
     1261 aacgaggcgc gctttccttt tttctttttg ctttttcttt ttttttctct tgaactcgac
     1321 ggatctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
     1381 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     1441 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     1501 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     1561 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     1621 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     1681 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     1741 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     1801 ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg
     1861 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     1921 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     1981 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta ccggccgcaa
     2041 attaaagcct tcgagcgtcc caaaaccttc tcaagcaagg ttttcagtat aatgttacat
     2101 gcgtacacgc gtctgtacag aaaaaaaaga aaaatttgaa atataaataa cgttcttaat
     2161 actaacataa ctataaaaaa ataaataggg acctagactt caggttgtct aactccttcc
     2221 ttttcggtta gagcggatgt ggggggaggg cgtgaatgta agcgtgacat aactaattac
     2281 atgactcgag ttaattaagc ttgtgcccca gtttgctagg gaggtcgcag tatctggcca
     2341 ctgccacctc gtgctgctcg acgtaggtct cgttgttggc ctccttgatt ctttccagtc
     2401 tgtagtccac atagtagacg ccaggcatct tgaggttctt agcgggtttc ttggatctat
     2461 atgtggtctt gatgtttgcg atcagatggc tcccgcccac gagcttcagg gccatgtcgt
     2521 ttctgccttc caggccgccg tcagcggggt acagcgtctc ggtgaaggcc tcccagccga
     2581 gtgttttctt ctgcatcaca gggccgttgg atgtgaagtt cacccctctg atcttgacgt
     2641 tgtagatgag gcagccgtcc tggaggctgg tgtcctgggt agcggtcagc acgcccccgt
     2701 cttcgtatgt ggtgactctc tcccatgtga agccctcagg gaaggactgc ttgaagaagt
     2761 cggggatgcc ctgggtgtgg ttgatgaagg tcttgctgcc gtagaggaag ctagtagcca
     2821 ggatgtcgaa ggcgaagggg agagggccgc cctcgaccac cttgattctc atggtctggg
     2881 tgccctcgta gggcttgcct tcgccctcgg atgtgcactt gaagtgatgg ttgtccacgg
     2941 tgccctccat gtacagcttc atgtgcatgt tctccttaat cagctcgctc atagatctgg
     3001 tacccgggga tccggaagag taggcgcgct tctggaaagc ctggcgggtg gtggcgttga
     3061 cgatggaggt catctgggtg cgcttgaggg tctggagggg ggagccagtc tggatggtgc
     3121 gcttgctgac ctgagcaacg cggacagcag ggcgggcaac cttggcggaa gcagccatct
     3181 gggaggccag gcgagaggcg aggacacgag tggaggccat actagttcta gaatccgtcg
     3241 aaactaagtt ctggtgtttt aaaactaaaa aaaagactaa ctataaaagt agaatttaag
     3301 aagtttaaga aatagattta cagaattaca atcaatacct accgtcttta tatacttatt
     3361 agtcaagtag gggaataatt tcagggaact ggtttcaacc ttttttttca gctttttcca
     3421 aatcagagag agcagaaggt aatagaaggt gtaagaaaat gagatagata catgcgtggg
     3481 tcaattgcct tgtgtcatca tttactccag gcaggttgca tcactccatt gaggttgtgc
     3541 ccgttttttg cctgtttgtg cccctgttct ctgtagttgc gctaagagaa tggacctatg
     3601 aactgatggt tggtgaagaa aacaatattt tggtgctggg attctttttt tttctggatg
     3661 ccagcttaaa aagcgggctc cattatattt agtggatgcc aggaataaac tgttcaccca
     3721 gacacctacg atgttatata ttctgtgtaa cccgccccct attttgggca tgtacgggtt
     3781 acagcagaat taaaaggcta attttttgac taaataaagt taggaaaatc actactatta
     3841 attatttacg tattctttga aatggcgagt attgataatg ataaactgag ctccaccgcg
     3901 gtggcggccg ctctagaact agtggatccc ccgggctgca ggaattcgat atcaagctta
     3961 tcgataccgt cgacctcgag ggggggcccg gtacccagct tttgttccct ttagtgaggg
     4021 ttaattccga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4081 ctcacaattc cacacaacat aggagccgga agcataaagt gtaaagcctg gggtgcctaa
     4141 tgagtgaggt aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4201 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4261 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4321 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4381 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4441 ctggcgtttt tccataggct cggcccccct gacgagcatc acaaaaatcg acgctcaagt
     4501 cagaggtggc gaaacccgac aggactataa agataccagg cgttcccccc tggaagctcc
     4561 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4621 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     4681 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4741 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4801 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4861 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     4921 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4981 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5041 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5101 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5161 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5221 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactg
     5281 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5341 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5401 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5461 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5521 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5581 caacgatcaa ggcgagttac atgatccccc atgttgtgaa aaaaagcggt tagctccttc
     5641 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5701 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5761 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5821 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5881 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5941 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6001 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6061 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6121 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6181 ccccgaaaag tgccacctgg gtccttttca tcacgtgcta taaaaataat tataatttaa
     6241 attttttaat ataaatatat aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa
     6301 atagtttttg ttttccgaag atgtaaaaga ctctaggggg atcgccaaca aatactacct
     6361 tttatcttgc tcttcctgct ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta
     6421 gaagaccaca cacgaaaatc ctgtgatttt acattttact tatcgttaat cgaatgtata
     6481 tctatttaat ctgcttttct tgtctaataa atatatatgt aaagtacgct ttttgttgaa
     6541 attttttaaa cctttgttta tttttttttc ttcattccgt aactcttcta ccttctttat
     6601 ttactttcta aaatccaaat acaaaacata aaaataaata aacacagagt aaattcccaa
     6661 attattccat cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg atccgtccta
     6721 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6781 tc
//
