LOCUS       Exported                9863 bp ds-DNA     circular SYN 05-JAN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pS-cr:GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9863)
  AUTHORS   Administrator
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jan 9, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9863
                     /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_feature    2969..2988
                     /label=LbCpf1 direct repeat
                     /note="LbCpf1 direct repeat"
     misc_feature    2989..3011
                     /label=BsaI
                     /note="BsaI"
     misc_feature    3012..3031
                     /label=LbCpf1 direct repeat
                     /note="LbCpf1 direct repeat"
     terminator      3224..3476
                     /label=Nos
                     /note="NOS T"
     misc_feature    3524..3548
                     /label=RB
                     /note="RB"
     rep_origin      3676..5018
                     /label=2u ori
                     /note="2u ori"
                     /note="yeast 2u plasmid origin of replication"
     promoter        5277..5558
                     /gene="S. cerevisiae TRP1"
                     /label=S. cerevisiae TRP1 promoter
                     /note="TRP1 promoter"
     CDS             5559..6233
                     /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(6614..7762)
                     /codon_start=1
                     /label=trfA
                     /note="trfA"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(8061..8855)
                     /codon_start=1
                     /label=KanR
                     /note="KanR"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      9067..9698
                     /label=oriV
                     /note="oriV"
     misc_feature    9776..9800
                     /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 atactagtat ctacacttag tagaaattgg agaccgaggt
     3001 ccgaggtctc catctacact tagtagaaat tactagttat ggttattggt actgtgttct
     3061 tattacagta ttgtgatttt ctaagtgagg tttgattaag aataaaatta tgacacaatt
     3121 gctctgggtc ggcatggcat ctccacctcc tcgcggtccg acctgggcat ccgaaggagg
     3181 acgtcgtcca ctcggatggc taagggagag ctcgaatttc cccgatcgtt caaacatttg
     3241 gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt
     3301 tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag
     3361 atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat
     3421 atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcggaa
     3481 ttcagattgt cgtttcccgc cttcagttta aactatcagt gtttgacagg atatattggc
     3541 gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa
     3601 aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acaggagatc tcagtaaagc
     3661 gaaaagtgcc acctgaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     3721 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3781 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     3841 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     3901 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     3961 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     4021 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     4081 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     4141 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     4201 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     4261 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     4321 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     4381 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     4441 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     4501 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     4561 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt
     4621 ggttttcaaa agcgctctga agttcctata ctttctagag aataggaact tcggaatagg
     4681 aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata
     4741 cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag
     4801 aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct atttatgtag
     4861 gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg
     4921 cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc
     4981 tcatccttca atgctatcat ttcctttgat attggatcat attaagaaac cattattatc
     5041 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     5101 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5161 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5221 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tagatcaacg
     5281 acattactat atatataata taggaagcat ttaatagaac agcatcgtaa tatatgtgta
     5341 ctttgcagtt atgacgccag atggcagtag tggaagatat tctttattga aaaatagctt
     5401 gtcaccttac gtacaatctt gatccggagc ttttcttttt ttgccgatta agaattaatt
     5461 cggtcgaaaa aagaaaagga gagggccaag agggagggca ttggtgacta ttgagcacgt
     5521 gagtatacgt gattaagcac acaaaggcag cttggagtat gtctgttatt aatttcacag
     5581 gtagttctgg tccattggtg aaagtttgcg gcttgcagag cacagaggcc gcagaatgtg
     5641 ctctagattc cgatgctgac ttgctgggta ttatatgtgt gcccaataga aagagaacaa
     5701 ttgacccggt tattgcaagg aaaatttcaa gtcttgtaaa agcatataaa aatagttcag
     5761 gcactccgaa atacttggtt ggcgtgtttc gtaatcaacc taaggaggat gttttggctc
     5821 tggtcaatga ttacggcatt gatatcgtcc aactgcatgg agatgagtcg tggcaagaat
     5881 accaagagtt cctcggtttg ccagttatta aaagactcgt atttccaaaa gactgcaaca
     5941 tactactcag tgcagcttca cagaaacctc attcgtttat tcccttgttt gattcagaag
     6001 caggtgggac aggtgaactt ttggattgga actcgatttc tgactgggtt ggaaggcaag
     6061 agagccccga aagcttacat tttatgttag ctggtggact gacgccagaa aatgttggtg
     6121 atgcgcttag attaaatggc gttattggtg ttgatgtaag cggaggtgtg gagacaaatg
     6181 gtgtaaaaga ctctaacaaa atagcaaatt tcgtcaaaaa tgctaagaaa taggttatta
     6241 ctgagtagta tttatttaag tattgtttgt gcacttgccg atctatgcgg tgtgaaatac
     6301 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaaacgtta atattttgtt
     6361 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     6421 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     6481 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtctc
     6541 ggtgcgggcc tcttcgctat tacgccagat ccttgctggc tgaaccccca gccggaactg
     6601 accccacaag gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca
     6661 ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca
     6721 cgcgggtgga atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct
     6781 cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact
     6841 tctcccatat gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga
     6901 tcaggacctg gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca
     6961 gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta
     7021 ggcgcgacag gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt
     7081 cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt
     7141 actccaacac ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt
     7201 aggtgatctt cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg
     7261 ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc
     7321 gcatcgtgtc cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct
     7381 gcttcgtgtg tttcagcaac gcggcctgct tggcctcgct gacctgtttt gccaggtcct
     7441 cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact
     7501 tcgccaaacc tgccgcctcc tgttcgagac gacgcgaacg ctccacggcg gccgatggcg
     7561 cgggcagggc agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct
     7621 ggaccatcga gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg
     7681 cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct
     7741 tccggtcaaa cgtccgattc attcaccctc cttgcgggat tgccccgact cacgccgggg
     7801 caatgtgccc ttattcctga tttgacccgc ctggtgcctt ggtgtccaga taatccacct
     7861 tatcggcaat gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac ttggtattcc
     7921 gaatcttgcc ctgcacgaat accagcgacc ccttgcccaa atacttgccg tgggcctcgg
     7981 cctgagagcc aaaacacttg atgcggaaga agtcggtgcg ctcctgcttg tcgccggcat
     8041 cgttgcgcca catctaggta ctaaaacaat tcatccagta aaatataata ttttattttc
     8101 tcccaatcag gcttgatccc cagtaagtca aaaaatagct cgacatactg ttcttccccg
     8161 atatcctccc tgatcgaccg gacgcagaag gcaatgtcat accacttgtc cgccctgccg
     8221 cttctcccaa gatcaataaa gccacttact ttgccatctt tcacaaagat gttgctgtct
     8281 cccaggtcgc cgtgggaaaa gacaagttcc tcttcgggct tttccgtctt taaaaaatca
     8341 tacagctcgc gcggatcttt aaatggagtg tcttcttccc agttttcgca atccacatcg
     8401 gccagatcgt tattcagtaa gtaatccaat tcggctaagc ggctgtctaa gctattcgta
     8461 tagggacaat ccgatatgtc gatggagtga aagagcctga tgcactccgc atacagctcg
     8521 ataatctttt cagggctttg ttcatcttca tactcttccg agcaaaggac gccatcggcc
     8581 tcactcatga gcagattgct ccagccatca tgccgttcaa agtgcaggac ctttggaaca
     8641 ggcagctttc cttccagcca tagcatcatg tccttttccc gttccacatc ataggtggtc
     8701 cctttatacc ggctgtccgt catttttaaa tataggtttt cattttctcc caccagctta
     8761 tataccttag caggagacat tccttccgta tcttttacgc agcggtattt ttcgatcagt
     8821 tttttcaatt ccggtgatat tctcatttta gccatttatt atttccttcc tcttttctac
     8881 agtatttaaa gataccccaa gaagctaatt ataacaagac gaactccaat tcactgttcc
     8941 ttgcattcta aaaccttaaa taccagaaaa cagctttttc aaagttgttt tcaaagttgg
     9001 cgtataacat agtatcgacg gagccgattt tgaaaccaca attatgggtg atgctgccaa
     9061 ctcgagagcg ggccgggagg gttcgagaag ggggggcacc ccccttcggc gtgcgcggtc
     9121 acgcgcacag ggcgcagccc tggttaaaaa caaggtttat aaatattggt ttaaaagcag
     9181 gttaaaagac aggttagcgg tggccgaaaa acgggcggaa acccttgcaa atgctggatt
     9241 ttctgcctgt ggacagcccc tcaaatgtca ataggtgcgc ccctcatctg tcagcactct
     9301 gcccctcaag tgtcaaggat cgcgcccctc atctgtcagt agtcgcgccc ctcaagtgtc
     9361 aataccgcag ggcacttatc cccaggcttg tccacatcat ctgtgggaaa ctcgcgtaaa
     9421 atcaggcgtt ttcgccgatt tgcgaggctg gccagctcca cgtcgccggc cgaaatcgag
     9481 cctgcccctc atctgtcaac gccgcgccgg gtgagtcggc ccctcaagtg tcaacgtccg
     9541 cccctcatct gtcagtgagg gccaagtttt ccgcgaggta tccacaacgc cggcggccgc
     9601 ggtgtctcgc acacggcttc gacggcgttt ctggcgcgtt tgcagggcca tagacggccg
     9661 ccagcccagc ggcgagggca accagcccgg tgagcgtcta gtggactgat gggctgcctg
     9721 tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag ctgttggctg gctggtggca
     9781 ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca ttgcggacgt
     9841 ttttaatgta ctggggtggt ttt
//
