LOCUS       Exported                9083 bp ds-DNA     circular SYN 23-JUN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9083)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jun 23, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9083
                     /organism="synthetic DNA construct"
                     /note="/dnas_title=pBV514  pUC19-LEU2-OsTirF74A-9Myc into 
                     TRP1
                     locus"
     misc_feature    2398..2414
                     /label=overhang
                     /note="overhang"
     promoter        2415..3039
                     /note="TRP1 5'UTR"
     gene            2415..2444
     gene            3013..3039
     misc_feature    3029..3049
                     /label=overhang
                     /note="overhang"
     promoter        complement(3040..3098)
                     /note="URA3pro"
     misc_feature    3040..3063
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
     misc_feature    3066..3091
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="Campr-ERG11-seq2"
     promoter        complement(3099..3745)
                     /note="LEU2pro"
     misc_feature    3728..3753
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="Campr-ERG11-seq3"
     gene            3746..4840
                     /label=LEU2
                     /note="LEU2"
     misc_feature    complement(3951)
                     /label=sense mutation V into A
                     /note="sense mutation V into A "
     misc_feature    4043..4067
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
     misc_feature    4043..4060
                     /label=overhang
                     /note="overhang"
     misc_feature    4367..4388
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="Campr-ERG11-seq4"
     misc_feature    complement(4642..4643)
                     /label=Sense mutation Y into D (gggtCAacc into gggtTGacc)
                     /note="Sense mutation Y into D (gggtCAacc into gggtTGacc)"
     terminator      complement(4841..5316)
                     /note="LEU2term"
     misc_feature    5016..5039
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="Campr-ERG11-seq5"
     misc_feature    complement(5298..5316)
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
     promoter        5317..6023
                     /note="ADH1pr"
     misc_feature    5317..5334
                     /label=overhang
                     /note="overhang"
     misc_feature    5605..5630
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="pUC19ostirseq1"
     CDS             6024..8137
                     /codon_start=1
                     /label=OsTir1-9Myc
                     /note="OsTir1-9Myc"
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHAADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKILV
                     DSGSAASGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGSSRGEQKLISEEDLN
                     GEQKLISEEDLNGEQKLISEEDLNGSSRGEQKLISEEDLNGEQKLISEEDLNGEQKLIS
                     EEDLNGSTS"
     misc_feature    6150..6169
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="pUC19ostirseq2"
     misc_feature    complement(6150..6169)
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="pUC19ostirseq2-R"
     misc_feature    complement(6227..6242)
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
     misc_feature    6243..6261
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
     misc_feature    6243..6256
                     /label=overhang
                     /note="overhang"
     misc_feature    6745..6770
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="pUC19ostirseq3"
     misc_feature    7329..7354
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="pUC19ostirseq4"
     misc_feature    complement(7659..7680)
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
     misc_feature    7663..7680
                     /label=overhang
                     /note="overhang"
     misc_feature    7764..7786
     misc_feature    7912..7935
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="pUC19ostirseq5"
     misc_feature    8126..8149
     misc_feature    8142..8162
                     /label=overhang
                     /note="overhang"
     misc_feature    8163..8820
                     /label=TRP1 3'UTR
                     /note="TRP1 3'UTR"
     misc_feature    8163..8189
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
     misc_feature    8349..8373
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="pUC19uwaCDR1GFPseq6"
     misc_feature    complement(8794..8820)
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
     misc_feature    8821..8837
                     /label=overhang
                     /note="overhang"
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     1981 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2041 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2101 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     2161 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     2221 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     2281 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     2341 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     2401 ttcgagctcg gtacttaatt aacgtcgctt tgagagaaat cagaagattc caaaagtcta
     2461 ctgaactttt gatcagaaag ttgcctttcc aaagactagt cagagaaatc gcccaagatt
     2521 tcaagaccga tctaagattc caatcctctg ccatcggtgc tctacaggaa tccgttgaag
     2581 cttacttggt ctctttgttc gaagacacca acttggccgc tatccacgct aagcgtgtta
     2641 ccatccaaaa gaaggatatc aagttggcta gaagattgag aggtgaaaga tcctaagcac
     2701 acagggattg ttgtacaatt actttttatg attttagata cacacaaatc ttttctacga
     2761 ttacctttca tttattttcg tttcatgttt tattaatcat tctttttgta tatttcacat
     2821 cgggtggggt ttttatatta aattttaata acataatctt ttaaagtttc tactttctat
     2881 cctgtatatt atttccatat ctatatctat attttcaagc ttgttcacat ccttatcgga
     2941 attgttccag cgtttttgta cagaaaatga tgttatggta acgagctata tcagacctac
     3001 tagtcttata gagctgtaca gagtcttgga aacataaccg tgttgaagaa acatgaaatt
     3061 gcccagtatt cttaacccaa ctgcacagaa caaaaacccc tcgaggagaa cttctagtat
     3121 atccacatac ctaatattat tgccttatta aaaatggaat cccaacaatt acatcaaaat
     3181 ccacattctc ttcaaaatca attgtcctgt acttccttgt tcatgtgtgt tcaaaaacgt
     3241 tatatttata ggataattat actctatttc tcaacaagta attggttgtt tggccgagcg
     3301 gtctaaggcg cctgattcaa gaaatatctt gaccgcagtt aactgtggga atactcaggt
     3361 atcgtaagat gcaagagttc gaatctctta gcaaccatta tttttttcct caacataacg
     3421 agaacacaca ggggcgctat cgcacagaat caaattcgat gactggaaat tttttgttaa
     3481 tttcagaggt cgcctgacgc atataccttt ttcaactgaa aaattgggag aaaaaggaaa
     3541 ggtgagaggc cggaaccggc ttttcatata gaatagagaa gcgttcatga ctaaatgctt
     3601 gcatcacaat acttgaagtt gacaatatta tttaaggacc tattgttttt tccaataggt
     3661 ggttagcaat cgtcttactt tctaactttt cttacctttt acatttcagc aatatatata
     3721 tatatttcaa ggatatacca ttctaatgtc tgcccctaag aagatcgtcg ttttgccagg
     3781 tgaccacgtt ggtcaagaaa tcacagccga agccattaag gttcttaaag ctatttctga
     3841 tgttcgttcc aatgtcaagt tcgatttcga aaatcattta attggtggtg ctgctatcga
     3901 tgctacaggt gttccacttc cagatgaggc gctggaagcc tccaagaagg ctgatgccgt
     3961 tttgttaggt gctgtgggtg gtcctaaatg gggtaccggt agtgttagac ctgaacaagg
     4021 tttactaaaa atccgtaaag aacttcaatt gtacgccaac ttaagaccat gtaactttgc
     4081 atccgactct cttttagact tatctccaat caagccacaa tttgctaaag gtactgactt
     4141 cgttgttgtc agagaattag tgggaggtat ttactttggt aagagaaagg aagacgatgg
     4201 tgatggtgtc gcttgggata gtgaacaata caccgttcca gaagtgcaaa gaatcacaag
     4261 aatggccgct ttcatggccc tacaacatga gccaccattg cctatttggt ccttggataa
     4321 agctaatgtt ttggcctctt caagattatg gagaaaaact gtggaggaaa ccatcaagaa
     4381 cgaattccct acattgaagg ttcaacatca attgattgat tctgccgcca tgatcctagt
     4441 taagaaccca acccacctaa atggtattat aatcaccagc aacatgtttg gtgatatcat
     4501 ctccgatgaa gcctccgtta tcccaggttc cttgggtttg ttgccatctg cgtccttggc
     4561 ctctttgcca gacaagaaca ccgcatttgg tttgtacgaa ccatgccacg gttctgctcc
     4621 agatttgcca aagaataagg tcaaccctat cgccactatc ttgtctgctg caatgatgtt
     4681 gaaattgtca ttgaacttgc ctgaagaagg taaggccatt gaagatgcag ttaaaaaggt
     4741 tttggatgca ggtatcagaa ctggtgattt aggtggttcc aacagtacca ccgaagtcgg
     4801 tgatgctgtc gccgaagaag ttaagaaaat ccttgcttaa aaagattctc tttttttatg
     4861 atatttgtac ataaacttta taaatgaaat tcataataga aacgacacga aattacaaaa
     4921 tggaatatgt tcatagggta gacgaaacta tatacgcaat ctacatacat ttatcaagaa
     4981 ggagaaaaag gaggatagta aaggaataca ggtaagcaaa ttgatactaa tggctcaacg
     5041 tgataaggaa aaagaattgc actttaacat taatattgac aaggaggagg gcaccacaca
     5101 aaaagttagg tgtaacagaa aatcatgaaa ctacgattcc taatttgata ttggaggatt
     5161 ttctctaaaa aaaaaaaaat acaacaaata aaaaacactc aatgacctga ccatttgatg
     5221 gagtttaagt caataccttc ttgaaccatt tcccataatg gtgaaagttc cctcaagaat
     5281 tttactctgt cagaaacggc cttacgacgt agtcgacggg tgtacaatat ggacttcctc
     5341 ttttctggca accaaaccca tacatcggga ttcctataat accttcgttg gtctccctaa
     5401 catgtaggtg gcggagggga gatatacaat agaacagata ccagacaaga cataatgggc
     5461 taaacaagac tacaccaatt acactgcctc attgatggtg gtacataacg aactaatact
     5521 gtagccctag acttgatagc catcatcata tcgaagtttc actacccttt ttccatttgc
     5581 catctattga agtaataata ggcgcatgca acttcttttc tttttttttc ttttctctct
     5641 cccccgttgt tgtctcacca tatccgcaat gacaaaaaaa tgatggaaga cactaaagga
     5701 aaaaattaac gacaaagaca gcaccaacag atgtcgttgt tccagagctg atgaggggta
     5761 tctcgaagca cacgaaactt tttccttcct tcattcacgc acactactct ctaatgagca
     5821 acggtatacg gccttccttc cagttacttg aatttgaaat aaaaaaaagt ttgctgtctt
     5881 gctatcaagt ataaatagac ctgcaattat taatcttttg tttcctcgtc attgttctcg
     5941 ttccctttct tccttgtttc tttttctgca caatatttca agctatacca agcatacaat
     6001 caactatctc atatacaact agaatgacgt acttcccgga ggaggtggtg gagcacatct
     6061 tcagcttcct gccggcgcag cgcgaccgca acacggtctc gctcgtctgc aaggtgtggt
     6121 acgagatcga gaggctgagc cgccgcggcg tcttcgtggg caactgctac gccgtgcgcg
     6181 ccggccgcgt cgccgcgcgg ttccccaacg tgcgggcgct cacggtgaag gggaagcccc
     6241 acgccgccga cttcaacctc gtgccccccg actggggcgg ctacgcgggg ccgtggatcg
     6301 aggcggccgc gaggggatgc cacggcctgg aggagctcag gatgaagcgg atggtggtgt
     6361 ccgacgagag cctcgagctg ctggctcgct cgttcccgcg gttcagggct cttgttctta
     6421 tcagctgcga ggggttcagc actgacgggc tagccgccgt cgcgagccat tgcaagcttc
     6481 tgagggagtt ggatttgcag gaaaatgaag tggaggatcg agggcctagg tggctttcct
     6541 gcttccctga ttcctgcaca tcacttgtct cattgaattt tgcctgcatc aaaggggagg
     6601 ttaatgctgg ttcactggag agacttgtta gcaggtcccc aaacctgcgg agtttgaggc
     6661 tgaatcgatc tgtatcggta gatacacttg caaagatact actgcgtacc cctaacttgg
     6721 aggatttggg gacagggaat ttgacagatg acttccaaac tgagtcctac tttaagctta
     6781 ccagtgctct ggagaaatgc aagatgttga ggagtttgtc tggattctgg gatgcttctc
     6841 ctgtttgcct gtcatttatc taccccctgt gtgctcaact gacaggattg aacttgagct
     6901 atgcccccac acttgatgct tctgacctta caaaaatgat tagccgctgt gtgaagctcc
     6961 aacgcctttg ggtactggat tgtatctcgg acaaaggctt gcaagtggtg gcctccagtt
     7021 gcaaagactt gcaagaactc agggtatttc catcagattt ctacgtagct ggttattctg
     7081 cagtgacaga ggagggactt gttgcagtat ccttgggctg tccaaaactg aactcactac
     7141 tgtacttctg tcaccaaatg actaatgctg cactagttac tgtcgccaag aactgtccaa
     7201 atttcacacg attcagactt tgtattcttg agccagggaa gcctgatgtt gtgacaagcc
     7261 aaccattaga tgaaggcttt ggagctattg ttcgtgagtg caagggatta caacgtttgt
     7321 caatatctgg tcttctcaca gacaaagttt tcatgtatat tgggaaatat gcaaaacaac
     7381 ttgagatgct ttctatagca tttgctggtg acagtgataa gggtatgatg catgttatga
     7441 atggatgcaa gaatttaagg aaactggaga taagagatag cccgtttggt gatgctgcac
     7501 tcttggggaa ttttgctagg tacgagacaa tgcgatccct ttggatgtca tcttgcaatg
     7561 tcacgttaaa ggggtgccaa gtccttgcgt caaagatgcc gatgctcaat gttgaggtca
     7621 taaatgagcg ggatggtagc aatgaaatgg aggaaaacca tggagatctg cctaaagtgg
     7681 agaaattata tgtgtaccgc acaactgctg gggcgaggga tgatgcacca aattttgtta
     7741 aaatcctagt cgactccggt tctgctgcta gtggtgaaca aaagttgatt tctgaagaag
     7801 atttgaacgg ggaacaaaag ctaatctccg aggaagactt gaacggtgaa caaaaattaa
     7861 tctcagaaga agacttgaac ggatcctcta gaggtgaaca aaagttgatt tctgaagaag
     7921 atttgaacgg tgaacaaaag ctaatctccg aggaagactt gaacggtgaa caaaaattaa
     7981 tctcagaaga agacttgaac ggatcctcta gaggtgaaca aaagttgatt tctgaagaag
     8041 atttgaacgg tgaacaaaag ctaatctccg aggaagactt gaacggtgaa caaaaattaa
     8101 tctcagaaga agacttgaac ggatccacta gctaaaaagt ggaacgatca ttcaagagat
     8161 ccgtctcaat gaggttttac aaacaatcct ataatacata tatgccactt agatatactt
     8221 tgaatactga atatactgac tcattatatt ctttaattgt agtttctcaa aaatattcat
     8281 acaggttagc tatgaattca tgctataatt tccattttgt tcatgtaaga tacacatagg
     8341 gataaaatgt gataactaat cagccaactt ccgggtaccg tttatattat gcgtacttgt
     8401 tatcatgcgt tgcgttattc accctctttt taaatcgtca gtcgaattcg taaacttctt
     8461 atataatatc aatattgata acatctgttt attaatcaaa acattcgata agtcggaatt
     8521 cattcaatgc ataaaaatct gtcaatactg aggaacaatt ggagtttcag taaaaagtct
     8581 tgcaaagcat gccaagttca gtgactgtct tacacatatt accacagaag tatcttcacc
     8641 aaatttagtc aattaatata agtggatgta attgttaaat agtatttcag gagcaacata
     8701 tgcgattcga atcttcacat ccaaatagcc tctcattcac ggcagtaaga aaatgtgcta
     8761 tttgcctcag tagagatgta tagtctacat aaaccgaata cttgatatag accgacatcc
     8821 ctctagagtc gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg
     8881 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     8941 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     9001 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     9061 gaggcggttt gcgtattggg cgc
//
