LOCUS       sequence-102367-        7595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFA6a BirA-heh2 linker-SSH1::HIS5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7595)
  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 7595)
  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: pJW1509
FEATURES             Location/Qualifiers
     source          1..7595
                     /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"
     promoter        4176..4617
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    4176..4293
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4588..4611
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             4620..5582
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             4620..5582
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
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 acggttggat ggacgcaaag aagtttaata
     4021 atcatattac atggcattac caccatatac atatccatat ctaatcttac ttatatgttg
     4081 tggaaatgta aagagcccca ttatcttagc ctaaaaaaac cttctctttg gaactttcag
     4141 taatacgctt aactgctcat tgctatattg aagtacggat tagaagccgc cgagcgggcg
     4201 acagccctcc gacggaagac tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg
     4261 aaacgcagat gtgcctcgcg ccgcactgct ccgaacaata aagattctac aatactagct
     4321 tttatggtta tgaagaggaa aaattggcag taacctggcc ccacaaacct tcaaattaac
     4381 gaatcaaatt aacaaccata ggatgataat gcgattagtt ttttagcctt atttctgggg
     4441 taattaatca gcgaagcgat gatttttgat ctattaacag atatataaat ggaaaagctg
     4501 cataaccact ttaactaata ctttcaacat tttcagtttg tattacttct tattcaaatg
     4561 tcataaaagt atcaacaaaa aattgttaat atacctctat actttaacgt caaggagaaa
     4621 tgaaggataa caccgtgcca ctgaaattga ttgccctgtt agcgaacggt gaatttcact
     4681 ctggcgagca gttgggtgaa acgctgggaa tgagccgggc ggctattaat aaacacattc
     4741 agacactgcg tgactggggc gttgatgtct ttaccgttcc gggtaaagga tacagcctgc
     4801 ctgagcctat ccagttactt aatgctaaac agatattggg tcagctggat ggcggtagtg
     4861 tagccgtgct gccagtgatt gactccacga atcagtacct tcttgatcgt atcggagagc
     4921 ttaaatcggg cgatgcttgc attgcagaat accagcaggc tggccgtggt cgccggggtc
     4981 ggaaatggtt ttcgcctttt ggcgcaaact tatatttgtc gatgttctgg cgtctggaac
     5041 aaggcccggc ggcggcgatt ggtttaagtc tggttatcgg tatcgtgatg gcggaagtat
     5101 tacgcaagct gggtgcagat aaagttcgtg ttaaatggcc taatgacctc tatctgcagg
     5161 atcgcaagct ggcaggcatt ctggtggagc tgactggcaa aactggcgat gcggcgcaaa
     5221 tagtcattgg agccgggatc aacatggcaa tgcgccgtgt tgaagagagt gtcgttaatc
     5281 aggggtggat cacgctgcag gaagcgggga tcaatctcga tcgtaatacg ttggcggcca
     5341 tgctaatacg tgaattacgt gctgcgttgg aactcttcga acaagaagga ttggcacctt
     5401 atctgtcgcg ctgggaaaag ctggataatt ttattaatcg cccagtgaaa cttatcattg
     5461 gtgataaaga aatatttggc atttcacgcg gaatagacaa acagggggct ttattacttg
     5521 agcaggatgg aataataaaa ccctggatgg gcggtgaaat atccctgcgt agtgcagaaa
     5581 aaccaccaga gtctcctcca caatctaagt cagatgggaa agcaacttct gctgatttaa
     5641 cttctgaact ggaaactgta gaagaacttc ataagaaaga ttcttcggat gacaaaccaa
     5701 gggtaaaaga gcttccaaaa ccagaattgc cgaatttaaa agtatcaaac gaatttttag
     5761 ctcaactaaa taaagagtta gctagtgcag ctacggagaa ttatgatcat tctataaaat
     5821 ccacggactt atcgtctatc agaatcgaaa cagaagagcc tgttggtcct tcaactggag
     5881 cagaaacaag aaatgaaagt gaggtgatgg agaatatcaa tttggaagtc caacctgagg
     5941 tgaaggaagc aaaagaagaa ctaacaaaaa tatcagaaac ttttgataat caagacgaag
     6001 aggatactag taggttatcg tcgaaaaaaa atatacgaag tccaaaaggt cgcaccaggc
     6061 acttcattgc caacaaaact aaaagaggga tagatattat gaaaccgttt attgcacatc
     6121 tcatgtctgg ttttcgtcta attgatatcg ttaagcctat cctaccgatt ttaccggaag
     6181 ttgagttacc ttttgaaaaa ttaccatttg atgacaagat tgtctatact atttttgcag
     6241 ggttaattta cctttttgca caattcccac ttgttggttt gcccaaagct accacaccta
     6301 atgttaatga ccccatttat tttctaagag gtgtatttgg gtgtgaacca agaaccttgc
     6361 tagaatttgg cctttttccc aacatttcta gtgggttaat cttacaatta ttagccggtt
     6421 tgaaagtaat taaagttaat ttcaagatcc aaagtgatag agaattattt caaagcttaa
     6481 caaaagtttt tgccattgtt cagtatgtga ttttgaccaa cattttcata tttgcaggtt
     6541 attttggtga tgatttatct gtggtacaga tcgggttgat caactttcaa ttggtcggtg
     6601 ctggcatatt caccactttg ttagctgaag tcatcgataa aggatttgga ttttcttctg
     6661 gagctatgat aatcaacact gtggtaatcg ccactaactt ggtggctgat acttttggtg
     6721 tttctcaaat taaagttggc gaggacgacc aaaccgaagc ccaaggtgcc ctaatcaatt
     6781 tgattcaagg tttaagatct aagcacaaga catttattgg cggcatcatc tctgcattca
     6841 acagagacta tctaccaaac ttgacaacta ctattatagt tttggcaatt gccattattg
     6901 tgtgctattt gcaaagtgtc cgtgtagaat tgccaatcag atcaacaaga gcccgtggta
     6961 caaacaatgt ttacccaatt aagctactat acactggctg tttatctgtt ttgttttctt
     7021 acaccatctt gttctacatc cacatttttg ctttcgtttt aattcaattg gttgccaaga
     7081 atgaaccaac tcacatcatc tgcaaaataa tgggccacta tgagaatgct aataatcttt
     7141 tagcagtccc aaccttccca ttatctcttt tggctccacc aacatctttc ttcaaaggtg
     7201 tcactcaaca acctttaacc ttcatcacct attccgcctt tatattggtt actggtattt
     7261 ggtttgctga taagtggcag gctatttcag gttcctctgc tcgtgatgta gccttggaat
     7321 tcaaagacca aggtatcact ttgatgggac gcagagaaca gaatgtcgct aaagaattga
     7381 acaaagtcat tccaatcgca gctgttacgg gtgcatccgt cttaagtttg atcaccgtta
     7441 tcggtgaatc tttgggtttg aaaggtaaag ctgctggtat agttgttggc attgctggtg
     7501 ggttttcttt gctagaagtc attaccattg aataccaaca aagtggcggt caaagtgctt
     7561 tgaatcaagt cttgggtgtt ccaggtgcta tgtaa
//
