LOCUS       sequence-102446-        7164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFA6a Osm1(M33A)-yoEGFP::LEU2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7164)
  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 7164)
  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: pJW1517
FEATURES             Location/Qualifiers
     source          1..7164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(74..93)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(75..93)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(180..197)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(351..939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(431..450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1110..1901,1902..1970))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1733..1752
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1971..2075)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2143..2161
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2199..2221)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2321..2340
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2421..2439
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2421..2438
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        2490..2891
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             join(4419..4421,4422..5132)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     terminator      5156..5343
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5211..5235)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 gtttaaacga gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg
       61 gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat
      121 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
      181 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      241 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
      301 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      361 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      421 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      481 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      541 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      601 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      661 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      721 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      781 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      841 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      901 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
      961 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1021 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1081 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1141 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1201 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     1261 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1321 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1381 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1441 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1501 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1561 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1621 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1681 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1741 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1801 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1861 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1921 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1981 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2041 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2101 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2161 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     2221 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     2281 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     2341 ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc
     2401 ttatgtatca tacacatacg atttaggtga cactatagaa cgcggccgcc agctgaagct
     2461 tcgtacgctg caggtcgacg gtaataggcg catgcaactt cttttctttt tttttctttt
     2521 ctctctcccc cgttgttgtc tcaccatatc cgcaatgaca aaaaaatgat ggaagacact
     2581 aaaggaaaaa attaacgaca aagacagcac caacagatgt cgttgttcca gagctgatga
     2641 ggggtatctc gaagcacacg aaactttttc cttccttcat tcacgcacac tactctctaa
     2701 tgagcaacgg tatacggcct tccttccagt tacttgaatt tgaaataaaa aaaagtttgc
     2761 tgtcttgcta tcaagtataa atagacctgc aattattaat cttttgtttc ctcgtcattg
     2821 ttctcgttcc ctttcttcct tgtttctttt tctgcacaat atttcaagct ataccaagca
     2881 tacaatcaac tatgattaga tctgtgagaa gggttttcat ttacgtctca atattcgtat
     2941 tgataatagt tttgaaaaga acattaagtg gcacagatca aacgtcagct aaacaaccag
     3001 tggtggtcat tggctctggt ttggcaggct taaccacaag taatcgtctc attagtaaat
     3061 acagaattcc tgttgtgctt ttggataagg cggcttctat tggtgggaat tctataaagg
     3121 cttctagtgg tattaatggt gctcacacag acactcaaca aaatttaaag gtaatggaca
     3181 ctcccgaatt gtttttgaaa gatactttgc attcggctaa aggcagaggg gttccatcac
     3241 tgatggataa gttgactaag gaatccaaga gtgctatcag gtggttgcaa acagaattcg
     3301 atttgaaatt agacctcctt gcgcaattgg gcggtcactc tgttccaagg acccatagat
     3361 cttctggcaa attaccacca ggttttgaaa tcgtgcaagc gttatcaaaa aaactaaagg
     3421 atatctcttc caaagattcc aatctcgtgc agattatgct aaacagtgaa gtagtggata
     3481 tcgagcttga taatcaaggt catgttactg gtgtagtata tatggacgag aacggaaacc
     3541 gtaaaatcat gaagtcacac catgtcgtgt tttgctcagg tggatttggt tactctaagg
     3601 aaatgttgaa agagtactca ccaaatttga ttcacttgcc aactactaat ggcaaacaga
     3661 ctacaggtga tggtcaaaaa atcctttcaa agttgggtgc cgaattgatt gatatggatc
     3721 aagtgcaggt acaccctacc ggcttcattg atccaaatga ccgtgaaaat aactggaagt
     3781 ttttggctgc agaggcattg aggggtttag gcggcatctt attgcatccc accactggaa
     3841 gaaggtttac aaatgaattg agcaccagag atacagtaac catggaaata cagtctaaat
     3901 gtccgaaaaa tgataataga gcacttttgg taatgagcga caaagtctac gagaactata
     3961 cgaataacat aaacttttat atgtccaaaa acttaatcaa aaaagtgtca atcaacgatc
     4021 tgatccgaca atatgaccta caaactacag cttctgaact ggtaactgaa ctgaagagct
     4081 attccgatgt taatactaag gatacgtttg ataggccatt gattatcaat gcctttgata
     4141 aagatatttc gactgaatca actgtttatg ttggggaagt tacaccagtt gttcatttca
     4201 caatgggtgg tgtgaaaatt aatgagaaat ctcaggtaat taagaaaaat tcggaaagcg
     4261 ttctatctaa tgggatattt gctgctggtg aagtttcggg tggtgttcat ggagccaaca
     4321 gattgggtgg atctagtttg ttagagtgtg ttgtctttgg aaagacagct gcggataaca
     4381 tagcaaaatt gtacggtgac ggtgctggtt taattaacat gtcaaaaggc gaggaactat
     4441 tcacaggtgt cgtaccaata ctggtagaat tggatggcga cgtaaacggg cataagtttt
     4501 ctgtgagtgg tgagggtgaa ggggacgcaa cttatggcaa acttacactt aagttcatct
     4561 gcaccacagg gaagctgcct gtcccatggc caactctagt tacaactctc acttacggag
     4621 tacaatgttt ttctcgatac ccagaccaca tgaaacaaca tgactttttc aaatctgcca
     4681 tgcctgaggg atatgtgcaa gagagaacta ttttctttaa agatgatggt aattacaaaa
     4741 ctcgtgctga agtgaagttt gaaggagata cacttgttaa tagaattgag ttaaagggta
     4801 ttgactttaa ggaagatggt aatatcttag gtcacaaatt ggaatacaac tacaattcac
     4861 ataacgttta catcatggca gataaacaga aaaacggcat aaaagttaat ttcaaaatta
     4921 gacataatat cgaagatggt tccgttcaac tcgctgatca ttatcaacaa aatacaccaa
     4981 tcggagatgg acctgttcta ttgccagaca accattactt atctacacag agcgccttgt
     5041 caaaagatcc taatgaaaag agagatcaca tggttttgtt agaatttgtc accgctgcgg
     5101 gtataacgtt gggtatggat gaattgtaca aataaggcgc gccacttcta aataagcgaa
     5161 tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa
     5221 attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct
     5281 gtaggtcagg ttgctttctc aggtatagta tgaggtcgct cttattgacc acacctctac
     5341 cggcagatcc gctagggata acagggtaat atagatctgt ttttaccaat agatagcggt
     5401 ttaatataag gtttcaacaa gaaatatttt gtgaaaatca aaagttaata gtttcactgt
     5461 cgaaaatatg aaagtttaag ttttcagttc atcccattgt agttatcaaa tttctgtgtt
     5521 tcccgtatat gtgtgctctg tattcatcgc tatcttgaaa ctggttatga tcgttgcgca
     5581 tgtgacctgt atcgctcaaa agagcccggc accggtaccg ggcaaaagcc cctagcaagc
     5641 taaaatgatt caactttcgt agtaatttgt tcaaccggtc acagtaaagt caatgactaa
     5701 cttttgaatc gaggatgtat ttccaattga aagcaatgta tatataaacc attgtgttga
     5761 agcttgtctt acagttttct tgtatcctcc cattgttcat atagtactta aagtaccata
     5821 atccttaaag aacaaacttg aaagagaata cactatacac aactaataaa taaagaataa
     5881 tcatggctgt gaccaagaca attgtagttc taccaggtga ccatgttggt caagaaatca
     5941 ctgaagaggc cattaaagtt ttaaatgcta ttcaggaatg tcgtccagac aaggtcaatt
     6001 tcaagtttga tcatcatttg atcggtggtg ctgcaattga tgccactggt gttccattac
     6061 cagacgaagc tttggaggcc tctaagaaag ctgatgccgt gcttctgggt gctgttggtg
     6121 gtccaaaatg gggtactggc gctgtcagac cagaacaagg tcttttgaaa atccgtaagg
     6181 agttgcaatt gtatgctaat ctaagaccat gtaattttgc atctgattcc ttactagatc
     6241 tatcgccatt gaagcctgaa attgcaagag gtacagattt cgtcgttgtt agagaactag
     6301 tgggtggtat ttatttcggg gagagaaaag aagatgaagg tgatggtgtc gcctgggata
     6361 gcgaaaagta ttctgtgccc gaagttcaaa gaatcactag aatggctgct ttcatggctc
     6421 tgcaacacaa cccaccttta cctatttggt ctctagataa agccaacgtt ctagcatcat
     6481 caagattatg gagaaaaaca gtggaagaga cgattaagaa tgaattccca caattgactg
     6541 ttcaacatca attgattgat tccgctgcta tgatcttggt taagaaccca actcatctaa
     6601 atggtattat tatcacaaac aatatgtttg gtgatattat ctccgatgaa gcctccgtga
     6661 tcccaggttc gttaggatta cttccatctg cctccttggc atctcttcca gataagaata
     6721 cggcatttgg tttatatgag ccatgtcatg gctctgctcc agacttacca aagggaaagg
     6781 ttaaccccgt agcaacaatc ctatctgctg ctatgatgtt gaagttgtct ctggacctgt
     6841 tcgaagaagg tgaaattata gaacaagcag tcaagaaagt gttggattct ggtatcagaa
     6901 cagcggatct aaagggtacc aactctacta ctgaggtcgg tgatgcagta gcgaaagctg
     6961 tcagggaact attagcttag acttagtaga tgtacagttg catacataag tctgaacaga
     7021 aagaagtaaa tatcatatct agttcggttt agaaatcagt gactatattg gttttgacca
     7081 gatcgcaatc cttgcaagca tttagtaatg actatcaaac cggtgaatca tctgataagg
     7141 ctatattagc ttgtgcattc gcat
//
