LOCUS       Exported                9899 bp ds-DNA     circular SYN 09-JAN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pS-sg:GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9899)
  AUTHORS   Administrator
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jan 12, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9899
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          2969..3067
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..767
                     /label=2x35S
                     /note="2x35S"
     CDS             919..1695
                     /codon_start=1
                     /label=N
                     /note="TSWV-N"
                     /translation="MSKVKLTKENIVALLTQGKDLEFEEDQNLVAFNFKTFCLENLDQI
                     KKMSIISCLTFLKNRQSIMKVIKQSDFTFGKITIKKTSDRIGATDMTFRRLDSLIRVRL
                     VEETGNSENLNTIKSKIASHPLIQAYGLPLDDAKSVRLAIMLGGSLPLIASVDSFEMIS
                     VVLAIYQDAKYKDLGIDPKKYDTREALGKVCTVLKSKAFEMNEDQVKKGKEYAAILSSS
                     NPNAKGSIAMEHYSETLNKFYEMFGVKKQTKLAELA"
     CDS             complement(2243..2962)
                     /codon_start=1
                     /label=eGFP
                     /note="eGFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_RNA        complement(2969..3044)
                     /label=gRNA scaffold
                     /note="gRNA scaffold"
                     /note="guide RNA scaffold for the CRISPR/Cas9 system"
     misc_feature    3045..3067
                     /label=BsaI
                     /note="BsaI"
     terminator      3260..3512
                     /label=Nos
                     /note="NOS T"
     misc_feature    3560..3584
                     /label=RB
                     /note="RB"
     rep_origin      3712..5054
                     /label=2u ori
                     /note="2u ori"
                     /note="yeast 2u plasmid origin of replication"
     promoter        5313..5594
                     /gene="S. cerevisiae TRP1"
                     /label=S. cerevisiae TRP1 promoter
                     /note="TRP1 promoter"
     CDS             5595..6269
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required
                     for tryptophan biosynthesis"
                     /label=S. cerevisiae TRP1
                     /note="TRP1"
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     CDS             complement(6650..7798)
                     /codon_start=1
                     /label=trfA
                     /note="trfA"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(8097..8891)
                     /codon_start=1
                     /label=KanR
                     /note="KanR"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      9103..9734
                     /label=oriV
                     /note="oriV"
     misc_feature    9812..9836
                     /label=LB
                     /note="LB"
ORIGIN
        1 aagcttgcat gcctgcagtc aacatggtgg agcacgacac tctcgtctac tccaagaata
       61 tcaaagatac agtctcagaa gaccagaggg ctattgagac ttttcaacaa agggtaatat
      121 cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcgaa aggacagtag
      181 aaaaggaaga tggcttctac aaatgccatc attgcgataa aggaaaggct atcgttcaaa
      241 gaatgcctct accgacagtg gtcccaaaga tggacccccc acccacgagg aacatcgtgg
      301 aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat aacatggtgg
      361 agcacgacac tctcgtctac tccaagaata tcaaagatac agtctcagaa gaccagaggg
      421 ctattgagac tttcaacaaa gggtaatatc gggaaacctc ctcggattcc attgcccagc
      481 tatctgtcac ttcatcgaaa ggacagtaga aaaggaagat ggcttctaca aatgccatca
      541 ttgcgataaa ggaaaggcta tcgttcaaga atgcctctac cgacagtggt cccaaagatg
      601 gacccccacc cacgaggaac atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc
      661 aagtggattg atgtgatatc tccactgacg taagggatga cgcacaatcc cactatcctt
      721 cgcaagaccc ttcctctata taaggaagtt catttcattt ggagaggaga gcaattgtgt
      781 caattttatt caaaccttaa cactcagtct tacaaatcat cacattaaaa ccctaagaaa
      841 cgactgcgga atacagagtt gtactttcgc accttgaatt acatacggtc aaagcatata
      901 acaacttcta cgatcatcat gtctaaggtt aagctcacta aggaaaacat tgttgctttg
      961 ttgacacaag gcaaagatct tgaatttgaa gaagatcaga atctggtagc atttaacttc
     1021 aagacttttt gtctggaaaa ccttgaccag atcaagaaga tgagcattat ttcatgtctg
     1081 acattcctga agaatcgtca gagtataatg aaggttatta agcaaagtga ttttactttt
     1141 ggcaaaatca ctataaagaa aacttcagac aggattggag ccactgacat gaccttcaga
     1201 aggcttgata gcttgatcag ggtcaggctt gtcgaggaaa ctgggaattc tgagaatctc
     1261 aatactatca aatctaagat tgcttctcac cctctgattc aagcctatgg attacctctt
     1321 gatgatgcaa agtctgtgag gcttgccata atgctgggag gtagcttacc tcttattgct
     1381 tcagttgata gctttgagat gatcagtgtt gtcttggcta tatatcagga tgcaaaatac
     1441 aaagacctcg ggatcgatcc aaagaagtat gacaccaggg aagccttagg aaaagtttgc
     1501 actgtgctaa aaagcaaagc atttgaaatg aatgaagatc aggtgaagaa agggaaagag
     1561 tatgctgcta tacttagctc cagcaatcct aatgctaaag gaagtattgc tatggaacat
     1621 tacagtgaaa ctcttaacaa gttctatgaa atgttcgggg ttaaaaaaca gacaaaactt
     1681 gcagaacttg cttaaaagca gttgtaagtt aaattataaa aaagcctata aatatataaa
     1741 gctttcttta tctttattgc ttgtgcttgc ttagtgtgtt aaatttaaaa taagtgtgtt
     1801 taattaaagt ttgctttctg tgtgttgtgc ttaataataa acaaaataac aaaaacaaca
     1861 aaaaacataa aataaaaata agaaaaaata aaataaaata aaataaaata aataaataaa
     1921 aataaaaata aaataaaaac aaaaaacaaa aaacaaaaaa caaaacaaaa cccaaatttg
     1981 gccaaaattg tccctttcgg gatctttttg ggtttttgtt ttttaatttt ttgttgtttt
     2041 tatttcattt tttgattttt gtttgatttt gtttcatttt ttgatttttg tttcattttg
     2101 attttatttt tatttttatt tttattttta tggtttctat taaacaggag aaattcaaaa
     2161 gaaatgacaa aacagagaaa taattatgag taaagaaaga aaataaacat aacataatta
     2221 ggaaaagctg gacacggcaa gattacttgt acagctcgtc catgccgaga gtgatcccgg
     2281 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
     2341 gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
     2401 gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga
     2461 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
     2521 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     2581 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     2641 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
     2701 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
     2761 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     2821 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     2881 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     2941 gctcctcgcc cttgctcacc atactagtgc accgactcgg tgccactttt tcaagttgat
     3001 aacggactag ccttatttta acttgctatt tctagctcta aaaccgagac cgaggtccga
     3061 ggtctccact agttatggtt attggtactg tgttcttatt acagtattgt gattttctaa
     3121 gtgaggtttg attaagaata aaattatgac acaattgctc tgggtcggca tggcatctcc
     3181 acctcctcgc ggtccgacct gggcatccga aggaggacgt cgtccactcg gatggctaag
     3241 ggagagctcg aatttccccg atcgttcaaa catttggcaa taaagtttct taagattgaa
     3301 tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt
     3361 aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc
     3421 gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt
     3481 atcgcgcgcg gtgtcatcta tgttactaga tcggaattca gattgtcgtt tcccgccttc
     3541 agtttaaact atcagtgttt gacaggatat attggcgggt aaacctaaga gaaaagagcg
     3601 tttattagaa taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg
     3661 tatgtgcatg ccaaccacag gagatctcag taaagcgaaa agtgccacct gaacgaagca
     3721 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     3781 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
     3841 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
     3901 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     3961 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     4021 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     4081 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     4141 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     4201 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     4261 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     4321 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     4381 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
     4441 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
     4501 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
     4561 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt
     4621 tttggttttt tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt
     4681 cctatacttt ctagagaata ggaacttcgg aataggaact tcaaagcgtt tccgaaaacg
     4741 agcgcttccg aaaatgcaac gcgagctgcg cacatacagc tcactgttca cgtcgcacct
     4801 atatctgcgt gttgcctgta tatatatata catgagaaga acggcatagt gcgtgtttat
     4861 gcttaaatgc gtacttatat gcgtctattt atgtaggatg aaaggtagtc tagtacctcc
     4921 tgtgatatta tcccattcca tgcggggtat cgtatgcttc cttcagcact accctttagc
     4981 tgttctatat gctgccactc ctcaattgga ttagtctcat ccttcaatgc tatcatttcc
     5041 tttgatattg gatcatatta agaaaccatt attatcatga cattaaccta taaaaatagg
     5101 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5161 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     5221 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     5281 gagcagattg tactgagagt gcaccataga tcaacgacat tactatatat ataatatagg
     5341 aagcatttaa tagaacagca tcgtaatata tgtgtacttt gcagttatga cgccagatgg
     5401 cagtagtgga agatattctt tattgaaaaa tagcttgtca ccttacgtac aatcttgatc
     5461 cggagctttt ctttttttgc cgattaagaa ttaattcggt cgaaaaaaga aaaggagagg
     5521 gccaagaggg agggcattgg tgactattga gcacgtgagt atacgtgatt aagcacacaa
     5581 aggcagcttg gagtatgtct gttattaatt tcacaggtag ttctggtcca ttggtgaaag
     5641 tttgcggctt gcagagcaca gaggccgcag aatgtgctct agattccgat gctgacttgc
     5701 tgggtattat atgtgtgccc aatagaaaga gaacaattga cccggttatt gcaaggaaaa
     5761 tttcaagtct tgtaaaagca tataaaaata gttcaggcac tccgaaatac ttggttggcg
     5821 tgtttcgtaa tcaacctaag gaggatgttt tggctctggt caatgattac ggcattgata
     5881 tcgtccaact gcatggagat gagtcgtggc aagaatacca agagttcctc ggtttgccag
     5941 ttattaaaag actcgtattt ccaaaagact gcaacatact actcagtgca gcttcacaga
     6001 aacctcattc gtttattccc ttgtttgatt cagaagcagg tgggacaggt gaacttttgg
     6061 attggaactc gatttctgac tgggttggaa ggcaagagag ccccgaaagc ttacatttta
     6121 tgttagctgg tggactgacg ccagaaaatg ttggtgatgc gcttagatta aatggcgtta
     6181 ttggtgttga tgtaagcgga ggtgtggaga caaatggtgt aaaagactct aacaaaatag
     6241 caaatttcgt caaaaatgct aagaaatagg ttattactga gtagtattta tttaagtatt
     6301 gtttgtgcac ttgccgatct atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     6361 accgcatcag gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     6421 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     6481 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     6541 cgtggactcc aacgtcaaag ggcgaaaaac cgtctcggtg cgggcctctt cgctattacg
     6601 ccagatcctt gctggctgaa cccccagccg gaactgaccc cacaaggccc tagcgtttgc
     6661 aatgcaccag gtcatcattg acccaggcgt gttccaccag gccgctgcct cgcaactctt
     6721 cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca
     6781 tgaggcggaa ggtttccagc ttgagcgggt acggctcccg gtgcgagctg aaatagtcga
     6841 acatccgtcg ggccgtcggc gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt
     6901 ggtcgccagc aaacagcacg acgatttcct cgtcgatcag gacctggcaa cgggacgttt
     6961 tcttgccacg gtccaggacg cggaagcggt gcagcagcga caccgattcc aggtgcccaa
     7021 cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct
     7081 tcgtgtaata ccggccattg atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga
     7141 aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc caacacctgc tgccacacca
     7201 gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt gatcttcacg tccttgttga
     7261 cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga
     7321 acagggcaga gcgggccgtg tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa
     7381 tatcgaacaa ggaaagctgc atttccttga tctgctgctt cgtgtgtttc agcaacgcgg
     7441 cctgcttggc ctcgctgacc tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg
     7501 tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt
     7561 cgagacgacg cgaacgctcc acggcggccg atggcgcggg cagggcaggg ggagccagtt
     7621 gcacgctgtc gcgctcgatc ttggccgtag cttgctggac catcgagccg acggactgga
     7681 aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat ggtttcggca tcctcggcgg
     7741 aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg gtcaaacgtc cgattcattc
     7801 accctccttg cgggattgcc ccgactcacg ccggggcaat gtgcccttat tcctgatttg
     7861 acccgcctgg tgccttggtg tccagataat ccaccttatc ggcaatgaag tcggtcccgt
     7921 agaccgtctg gccgtccttc tcgtacttgg tattccgaat cttgccctgc acgaatacca
     7981 gcgacccctt gcccaaatac ttgccgtggg cctcggcctg agagccaaaa cacttgatgc
     8041 ggaagaagtc ggtgcgctcc tgcttgtcgc cggcatcgtt gcgccacatc taggtactaa
     8101 aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt gatccccagt
     8161 aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgat cgaccggacg
     8221 cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc aataaagcca
     8281 cttactttgc catctttcac aaagatgttg ctgtctccca ggtcgccgtg ggaaaagaca
     8341 agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg atctttaaat
     8401 ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca gatcgttatt cagtaagtaa
     8461 tccaattcgg ctaagcggct gtctaagcta ttcgtatagg gacaatccga tatgtcgatg
     8521 gagtgaaaga gcctgatgca ctccgcatac agctcgataa tcttttcagg gctttgttca
     8581 tcttcatact cttccgagca aaggacgcca tcggcctcac tcatgagcag attgctccag
     8641 ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc cagccatagc
     8701 atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct gtccgtcatt
     8761 tttaaatata ggttttcatt ttctcccacc agcttatata ccttagcagg agacattcct
     8821 tccgtatctt ttacgcagcg gtatttttcg atcagttttt tcaattccgg tgatattctc
     8881 attttagcca tttattattt ccttcctctt ttctacagta tttaaagata ccccaagaag
     8941 ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac cttaaatacc
     9001 agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta tcgacggagc
     9061 cgattttgaa accacaatta tgggtgatgc tgccaactcg agagcgggcc gggagggttc
     9121 gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt
     9181 taaaaacaag gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc
     9241 cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa
     9301 atgtcaatag gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg
     9361 cccctcatct gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca
     9421 ggcttgtcca catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg
     9481 aggctggcca gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg
     9541 cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca
     9601 agttttccgc gaggtatcca caacgccggc ggccgcggtg tctcgcacac ggcttcgacg
     9661 gcgtttctgg cgcgtttgca gggccataga cggccgccag cccagcggcg agggcaacca
     9721 gcccggtgag cgtctagtgg actgatgggc tgcctgtatc gagtggtgat tttgtgccga
     9781 gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat
     9841 tgacgcttag acaacttaat aacacattgc ggacgttttt aatgtactgg ggtggtttt
//
