LOCUS       sequence-112201-        6013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFA6a pSEC63-BirA-mVenus-UBC6-TA::HIS5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6013)
  AUTHORS   Jan CH, Williams CC, Weissman JS
  TITLE     Principles of ER cotranslational translocation revealed by 
            proximity-specific ribosome profiling.
  JOURNAL   Science. 2014 Nov 7;346(6210):1257521. doi: 10.1126/science.1257521.
            Epub 2014 Nov 6.
  PUBMED    25378630
REFERENCE   2  (bases 1 to 6013)
  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."; 
            date: "2014-11-7"; volume: "346(6210):1257521. doi"; pages: " 
            10.1126/science.1257521. Epub 2014 Nov 6"
COMMENT     Sequence Label: pJW1510
FEATURES             Location/Qualifiers
     source          1..6013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      21..208
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(76..100)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            283..1483
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        283..626
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             627..1280
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      1286..1483
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1587..1606)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1588..1606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1693..1710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1864..2452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1944..1963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2623..3414,3415..3483))
                     /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     3246..3265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3484..3588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3656..3674
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3712..3734)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3834..3853
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3934..3952
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3934..3951
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4253..5215
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             4253..5215
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             5231..5944
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             5231..5944
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
ORIGIN
        1 ggcgcgccac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
       61 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
      121 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg
      181 tcgctcttat tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga
      241 tctgtttagc ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat
      301 accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac
      361 atttagccca tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg
      421 gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc
      481 acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat
      541 ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct
      601 cacatcacat ccgaacataa acaaccatgg gtaggagggc ttttgtagaa agaaatacga
      661 acgaaacgaa aatcagcgtt gccatcgctt tggacaaagc tcccttacct gaagagtcga
      721 attttattga tgaacttata acttccaagc atgcaaacca aaagggagaa caagtaatcc
      781 aagtagacac gggaattgga ttcttggatc acatgtatca tgcactggct aaacatgcag
      841 gctggagctt acgactttac tcaagaggtg atttaatcat cgatgatcat cacactgcag
      901 aagatactgc tattgcactt ggtattgcat tcaagcaggc tatgggtaac tttgccggcg
      961 ttaaaagatt tggacatgct tattgtccac ttgacgaagc tctttctaga agcgtagttg
     1021 acttgtcggg acggccctat gctgttatcg atttgggatt aaagcgtgaa aaggttgggg
     1081 aattgtcctg tgaaatgatc cctcacttac tatattcctt ttcggtagca gctggaatta
     1141 ctttgcatgt tacctgctta tatggtagta atgaccatca tcgtgctgaa agcgctttta
     1201 aatctctggc tgttgccatg cgcgcggcta ctagtcttac tggaagttct gaagtcccaa
     1261 gcacgaaggg agtgttgtaa agagtactga caataaaaag attcttgttt tcaagaactt
     1321 gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat
     1381 ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt
     1441 aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta
     1501 acgccgccat ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc cactagtggc
     1561 ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt cgataagcca
     1621 ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1681 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1741 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1801 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1861 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1921 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1981 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2041 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2101 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2161 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2221 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2281 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2341 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2401 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2461 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2521 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2581 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2641 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2701 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2761 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2821 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2881 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2941 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3001 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3061 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3601 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     3661 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     3721 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     3781 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     3841 agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac gcggctacaa
     3901 ttaatacata accttatgta tcatacacat acgatttagg tgacactata gaacgcggcc
     3961 gccagctgaa gcttcgtacg ctgcaggtcg acggggtgct ctcatgcttc ttttctattt
     4021 tgagttatta acgtcttagc tttataatgt cacaatctga cttcattact gacacaatcg
     4081 ctcttataac gcgaattggt cactcgttac ggaaaggcgt tttctgaaaa atttgtcttg
     4141 aacacgtcga agatcactga acgaataaaa gatgcgactg gaacaatagt cagttataat
     4201 tgaccaaaga aggccttcgc aacaaaataa atatactgca tattacagca caatgaagga
     4261 taacaccgtg ccactgaaat tgattgccct gttagcgaac ggtgaatttc actctggcga
     4321 gcagttgggt gaaacgctgg gaatgagccg ggcggctatt aataaacaca ttcagacact
     4381 gcgtgactgg ggcgttgatg tctttaccgt tccgggtaaa ggatacagcc tgcctgagcc
     4441 tatccagtta cttaatgcta aacagatatt gggtcagctg gatggcggta gtgtagccgt
     4501 gctgccagtg attgactcca cgaatcagta ccttcttgat cgtatcggag agcttaaatc
     4561 gggcgatgct tgcattgcag aataccagca ggctggccgt ggtcgccggg gtcggaaatg
     4621 gttttcgcct tttggcgcaa acttatattt gtcgatgttc tggcgtctgg aacaaggccc
     4681 ggcggcggcg attggtttaa gtctggttat cggtatcgtg atggcggaag tattacgcaa
     4741 gctgggtgca gataaagttc gtgttaaatg gcctaatgac ctctatctgc aggatcgcaa
     4801 gctggcaggc attctggtgg agctgactgg caaaactggc gatgcggcgc aaatagtcat
     4861 tggagccggg atcaacatgg caatgcgccg tgttgaagag agtgtcgtta atcaggggtg
     4921 gatcacgctg caggaagcgg ggatcaatct cgatcgtaat acgttggcgg ccatgctaat
     4981 acgtgaatta cgtgctgcgt tggaactctt cgaacaagaa ggattggcac cttatctgtc
     5041 gcgctgggaa aagctggata attttattaa tcgcccagtg aaacttatca ttggtgataa
     5101 agaaatattt ggcatttcac gcggaataga caaacagggg gctttattac ttgagcagga
     5161 tggaataata aaaccctgga tgggcggtga aatatccctg cgtagtgcag aaaaagctgg
     5221 tttaattaac atgtctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga
     5281 attagatggt gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc
     5341 tacttacggt aaattgacct taaaattgat ttgtactact ggtaaattgc cagttccatg
     5401 gccaacctta gtcactactt taggttatgg tttgcaatgt tttgctagat acccagatca
     5461 tatgaaacaa catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac
     5521 tatttttttc aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga
     5581 taccttagtt aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt
     5641 aggtcacaaa ttggaataca actataactc tcacaatgtt tacatcactg ctgacaaaca
     5701 aaagaatggt atcaaagcta acttcaaaat tagacacaac attgaagatg gtggtgttca
     5761 attagctgac cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga
     5821 caaccattac ttatcctatc aatctaagtt atccaaagat ccaaacgaaa agagagacca
     5881 catggtcttg ttagaatttg ttactgctgc tggtattacc catggtatgg atgaattgta
     5941 caaaaacagc cgcgtgatgg tttatattgg tatcgctatt tttttgtttt tggttggcct
     6001 ttttatgaaa tga
//
