LOCUS       Exported                7043 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  OsTIR1 expression cassette marked with URA.
ACCESSION   .
VERSION     .
KEYWORDS    pJW1661
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7043)
  AUTHORS   Costa EA, Subramanian K, Nunnari J, Weissman JS
  TITLE     Defining the physiological role of SRP in protein-targeting 
            efficiency and specificity.
  JOURNAL   Science. 2018 Jan 18. pii: science.aar3607. doi: 
            10.1126/science.aar3607.
  PUBMED    29348368
REFERENCE   2  (bases 1 to 7043)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7043
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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        194..409
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             410..1213
                     /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(1344..1799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1481..1502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1693..1712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1925..1947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1939..1956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1940..1956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1963..1982
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1963..1981
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1990..2097
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2014..2030
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2014..2030
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2064..2080)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2065..2081)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2109..2127)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2148..2164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2148..2164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2161..2183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2172..2188
                     /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(2196..2226)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2241..2262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2379..2396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2550..3138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2630..2649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3309..4169)
                     /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     3932..3951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4170..4274)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4342..4360
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4661..5057
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             5070..6797
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein"
                     /label=OsTIR1
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     terminator      6854..7041
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6909..6933)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1921 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag
     1981 ggcgaattgg agctccaccg cggtggcggc cgctctagaa ctagtggatc ccccgggctg
     2041 caggaattcg atatcaagct tatcgatacc gtcgacctcg agggggggcc cggtaccagc
     2101 ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt
     2161 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2221 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2281 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2341 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2401 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2461 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2521 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2581 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2641 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2701 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2761 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2821 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2881 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2941 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3001 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3061 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3121 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3181 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3241 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3301 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3361 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3421 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3481 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3541 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3601 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3661 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3721 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3781 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3841 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3901 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3961 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4021 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4081 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4141 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4201 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4261 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     4321 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtcg ggtgtacaat
     4381 atggacttcc tcttttctgg caaccaaacc catacatcgg gattcctata ataccttcgt
     4441 tggtctccct aacatgtagg tggcggaggg gagatataca atagaacaga taccagacaa
     4501 gacataatgg gctaaacaag actacaccaa ttacactgcc tcattgatgg tggtacataa
     4561 cgaactaata ctgtagccct agacttgata gccatcatca tatcgaagtt tcactaccct
     4621 ttttccattt gccatctatt gaagtaataa taggcgcatg caacttcttt tctttttttt
     4681 tcttttctct ctcccccgtt gttgtctcac catatccgca atgacaaaaa aatgatggaa
     4741 gacactaaag gaaaaaatta acgacaaaga cagcaccaac agatgtcgtt gttccagagc
     4801 tgatgagggg tatctcgaag cacacgaaac tttttccttc cttcattcac gcacactact
     4861 ctctaatgag caacggtata cggccttcct tccagttact tgaatttgaa ataaaaaaaa
     4921 gtttgctgtc ttgctatcaa gtataaatag acctgcaatt attaatcttt tgtttcctcg
     4981 tcattgttct cgttcccttt cttccttgtt tctttttctg cacaatattt caagctatac
     5041 caagcataca atcaactatc tcatatacaa tgacgtactt cccggaggag gtggtggagc
     5101 acatcttcag cttcctgccg gcgcagcgcg accgcaacac ggtctcgctc gtctgcaagg
     5161 tgtggtacga gatcgagagg ctgagccgcc gcggcgtctt cgtgggcaac tgctacgccg
     5221 tgcgcgccgg ccgcgtcgcc gcgcggttcc ccaacgtgcg ggcgctcacg gtgaagggga
     5281 agccccactt cgccgacttc aacctcgtgc cccccgactg gggcggctac gcggggccgt
     5341 ggatcgaggc ggccgcgagg ggatgccacg gcctggagga gctcaggatg aagcggatgg
     5401 tggtgtccga cgagagcctc gagctgctgg ctcgctcgtt cccgcggttc agggctcttg
     5461 ttcttatcag ctgcgagggg ttcagcactg acgggctagc cgccgtcgcg agccattgca
     5521 agcttctgag ggagttggat ttgcaggaaa atgaagtgga ggatcgaggg cctaggtggc
     5581 tttcctgctt ccctgattcc tgcacatcac ttgtctcatt gaattttgcc tgcatcaaag
     5641 gggaggttaa tgctggttca ctggagagac ttgttagcag gtccccaaac ctgcggagtt
     5701 tgaggctgaa tcgatctgta tcggtagata cacttgcaaa gatactactg cgtaccccta
     5761 acttggagga tttggggaca gggaatttga cagatgactt ccaaactgag tcctacttta
     5821 agcttaccag tgctctggag aaatgcaaga tgttgaggag tttgtctgga ttctgggatg
     5881 cttctcctgt ttgcctgtca tttatctacc ccctgtgtgc tcaactgaca ggattgaact
     5941 tgagctatgc ccccacactt gatgcttctg accttacaaa aatgattagc cgctgtgtga
     6001 agctccaacg cctttgggta ctggattgta tctcggacaa aggcttgcaa gtggtggcct
     6061 ccagttgcaa agacttgcaa gaactcaggg tatttccatc agatttctac gtagctggtt
     6121 attctgcagt gacagaggag ggacttgttg cagtatcctt gggctgtcca aaactgaact
     6181 cactactgta cttctgtcac caaatgacta atgctgcact agttactgtc gccaagaact
     6241 gtccaaattt cacacgattc agactttgta ttcttgagcc agggaagcct gatgttgtga
     6301 caagccaacc attagatgaa ggctttggag ctattgttcg tgagtgcaag ggattacaac
     6361 gtttgtcaat atctggtctt ctcacagaca aagttttcat gtatattggg aaatatgcaa
     6421 aacaacttga gatgctttct atagcatttg ctggtgacag tgataagggt atgatgcatg
     6481 ttatgaatgg atgcaagaat ttaaggaaac tggagataag agatagcccg tttggtgatg
     6541 ctgcactctt ggggaatttt gctaggtacg agacaatgcg atccctttgg atgtcatctt
     6601 gcaatgtcac gttaaagggg tgccaagtcc ttgcgtcaaa gatgccgatg ctcaatgttg
     6661 aggtcataaa tgagcgggat ggtagcaatg aaatggagga aaaccatgga gatctgccta
     6721 aagtggagaa attatatgtg taccgcacaa ctgctggggc gagggatgat gcaccaaatt
     6781 ttgttaaaat cctatgatag gcctatccct aaccctctcc tcggtctcga ttctacgtaa
     6841 cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
     6901 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt
     6961 cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg aggtcgctct
     7021 tattgaccac acctctaccg gca
//
