LOCUS       40924_48158        8247 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast TAP expression vector pYL435, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8247)
  AUTHORS   Liu Y, Dinesh-Kumar SP.
  TITLE     A alternative yeast tandem affinity purification (TAP) expression 
            vector pYL435
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8247)
  AUTHORS   Liu Y, Dinesh-Kumar SP.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-AUG-2004) MCDB, Yale University, New Haven, CT 06520, 
            USA
REFERENCE   3  (bases 1 to 8247)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8247)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-AUG-2004) MCDB, Yale University, New Haven, CT 06520, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..443
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     promoter        475..493
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    511..635
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        809..911
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             912..1568
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1913..2215
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2259..2383)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2403..2474
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags"
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             2508..2525
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             2526..2549
                     /codon_start=1
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
                     /translation="LEVLFQGP"
     CDS             2589..2762
                     /codon_start=1
                     /label=ProtA
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2766..2936
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     terminator      2993..3240
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(3488..4076)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4250..5107)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(5206..6006)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6007..6227)
                     /label=URA3 promoter
     rep_origin      complement(6826..7706)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(7717..8230)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
       61 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
      121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
      181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
      241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
      301 taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc
      361 ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac
      421 ctctatactt taacgtcaag gagaaaaaac cccggatcgg actactagca gctgtaatac
      481 gactcactat agggaatatt aagctatcaa acaagtttgt acaaaaaagc tgaacgagaa
      541 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
      601 taatactgta aaacacaaca tatccagtca ctatggcggc cgcgggtgat gctgccaact
      661 tagcggccgc taagttggca gcatcacccg acgcactttg cgccgaataa atacctgtga
      721 cggaagatca cttcgcagaa taaataaatc ctggtgtccc tgttgatacc gggaagccct
      781 gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc aactttcacc
      841 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
      901 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
      961 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     1021 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     1081 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa
     1141 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     1201 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     1261 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     1321 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
     1381 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     1441 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtctgtg
     1501 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     1561 gcggggcgta atctagagga tccggcttac taaaagccag ataacagtat gcgtatttgc
     1621 gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa
     1681 gaggtgtgct atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct
     1741 caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc
     1801 cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc
     1861 cggtttattg aaatgaacgg ctcttttgct gacgagaaca gggactggtg aaatgcagtt
     1921 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
     1981 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
     2041 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
     2101 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
     2161 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
     2221 gtcaggctcc gttatacaca gccagtctgc aggtcgacca tagtgactgg atatgttgtg
     2281 ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta
     2341 tatcatttta cgtttctcgt tcagctttct tgtacaaagt ggttgagctc aagcttctcg
     2401 aggattacaa ggatgacgac gataaggact ataaagatga cgatgacaag gattacaaag
     2461 acgatgacga taagcctagg gattacgata tcccaacgac cgccagtcat caccatcacc
     2521 atcacctgga agttctgttc caggggcccg agcttaagac ggccgcccta gcgcaacacg
     2581 atgaagccgt agacaacaaa ttcaacaaag aacaacaaaa cgcgttctat gagatcttac
     2641 atttacctaa cttaaacgaa gaacaacgaa acgccttcat ccaaagttta aaagatgacc
     2701 caagccaaag cgctaacctt ttagcagaag ctaaaaagct aaatgatgct caggcgccga
     2761 aagtagacaa caaattcaac aaagaacaac aaaacgcgtt ctatgagatc ttacatttac
     2821 ctaacttaaa cgaagaacaa cgaaacgcct tcatccaaag tttaaaagat gacccaagcc
     2881 aaagcgctaa ccttttagca gaagctaaaa agctaaatga tgctcaggcg ccgaaagtag
     2941 acgcgaattc gagctcggta tgacccaaac ccgctgatcc tagagggccg catcatgtaa
     3001 ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa
     3061 ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta
     3121 agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt
     3181 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc
     3241 aagctgcggc cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3301 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3361 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3421 gaaagaacat gtgagcaaaa ggccagcaaa agcccaggaa ccgtaaaaag gccgcgttgc
     3481 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3541 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3601 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3661 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3721 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3781 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3841 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3901 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     3961 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4021 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4081 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4141 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4201 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4261 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4321 ccgtcgtgta gataactacg atacgggagc gcttaccatc tggccccagt gctgcaatga
     4381 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4441 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4501 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttggcattg
     4561 ctacaggcat cgtggtgtca ctctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4621 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4681 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4741 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4801 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4861 caatacggga taatagtgta tcacatagca gaactttaaa agtgctcatc attggaaaac
     4921 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4981 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5041 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5101 tactcatact cttccttttt caatgggtaa taactgatat aattaaattg aagctctaat
     5161 ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt gctggccgca
     5221 tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc
     5281 ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac
     5341 atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc
     5401 gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat
     5461 aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc
     5521 tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc
     5581 ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc
     5641 gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa
     5701 tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt
     5761 ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag tcaagatatc
     5821 cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact ccagtaattc
     5881 cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt gcatgatatt
     5941 aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat atgtagcttt
     6001 cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg ttaagaatac
     6061 tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg
     6121 ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaagaaa
     6181 ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aattgaaaag ctagcttatc
     6241 gatgataagc tgtcaaagat gagaattaat tccacggact atagactata ctagatactc
     6301 cgtctactgt acgatacact tccgctcagg tccttgtcct ttaacgaggc cttaccactc
     6361 ttttgttact ctattgatcc agctcagcaa aggcagtgtg atctaagatt ctatcttcgc
     6421 gatgtagtaa aactagctag accgagaaag agactagaaa tgcaaaaggc acttctacaa
     6481 tggctgccat cattattatc cgatgtgacg ctgcagcttc tcaatgatat tcgaatacgc
     6541 tttgaggaga tacagcctaa tatccgacaa actgttttac agatttacga tcgtacttgt
     6601 tacccatcat tgaattttga acatccgaac ctgggagttt tccctgaaac agatagtata
     6661 tttgaacctg tataataata tatagtctag cgctttacgg aagacaatgt atgtatttcg
     6721 gttcctggag aaactattgc atctattgca taggtaatct tgcacgtcgc atccccggtt
     6781 cattttctgc gtttccatct tgcacttcaa tagcatatct ttgttaacga agcatctgtg
     6841 cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     6901 agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc
     6961 tgtgcttcat ttttgtaaaa caaaaatgca acgcgacgag agcgctaatt tttcaaacaa
     7021 agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac
     7081 aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta
     7141 acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat
     7201 aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct
     7261 cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg
     7321 gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca
     7381 tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac
     7441 ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt
     7501 tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag
     7561 agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg
     7621 atgggtaggt tatataggga tatagcacag agatatatag caaagagata cttttgagca
     7681 atgtttgtgg aagcggtatt cgcaatggga agctccaccc cggttgataa tcagaaaagc
     7741 cccaaaaaca ggaagattgt ataagcaaat atttaaattg taaacgttaa tattttgtta
     7801 aaattcgcgt taaatttttg ttaaatcagc tcatttttta acgaatagcc cgaaatcggc
     7861 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttcc
     7921 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aagggtctat
     7981 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     8041 cgtaaagcag taaatcggaa gggtaaacgg atgcccccat ttagagcttg acggggaaag
     8101 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggggc tagggcggtg
     8161 ggaagtgtag gggtcacgct gggcgtaacc accacacccg ccgcgcttaa tggggcgcta
     8221 cagggcgcgt ggggatgatc cactagt
//
