LOCUS       pTK298                 11125 bp    DNA     circular UNA 31-JUL-2014
DEFINITION  SIN SP6 P1234 (nsP2 P726S) SGP(14) Kozak mKate-G8-L7Ae-PEST 4xFF4.
ACCESSION   urn.local.tasuku.1393282807282.539
VERSION     urn.local.tasuku.1393282807282.539
KEYWORDS    .
SOURCE      Sindbis
  ORGANISM  Sindbis
            .
FEATURES             Location/Qualifiers
     5'UTR           1..59
     CDS             60..1679
                     /label="nsP1"
     CDS             1680..4100
                     /label="nsP2"
     primer_bind     2517..2538
                     /label="TK-SIN-ClaI-ncp-seq-F1"
     misc_feature    2992
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    3579
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     Mutation        3855
                     /label="P726S ncp mutation"
     CDS             4101..5747
                     /label="nsP3"
     primer_bind     complement(4443..4462)
                     /label="TK-SIN-AvrII-ncp-seq-R1"
     Stop_codon      5748..5750
                     /label="Opal stop codon"
     CDS             5751..7601
                     /label="nsP4"
     primer_bind     6086..6105
                     /label="TK-SIN-ts-seq-F1"
     primer_bind     7002..7021
                     /label="TK-nsP4-SapI-UP-seq-F"
     primer_bind     7103..7126
                     /label="TK-nsP4-SapI-F"
     promoter        7500..7611
                     /label="Subgenomic promoter (98/14)"
     promoter        7579..7602
                     /label="Subgenomic promoter (19/5 minimal)"
     primer_bind     7596..7639
                     /label="TK-SIN-SGP14-Kozak-mKate-F"
     misc_feature    7598..7611
                     /label="Subgenomic RNA 5' UTR"
     misc_RNA        7612..7617
                     /label="Kozak"
     CDS             7618..8328
                     /label="mKate"
     CDS             8329..8352
                     /label="G8"
     CDS             8353..8709
                     /label="L7Ae"
     CDS             8710..8835
                     /label="PEST"
     misc_RNA        8850..8871
                     /label="FF4"
     misc_RNA        8872..8893
                     /label="FF4"
     misc_RNA        8894..8915
                     /label="FF4"
     misc_RNA        8916..8937
                     /label="FF4"
     primer_bind     complement(8940..8963)
                     /label="TK-3'UTR-XhoI-FF4-R"
     3'UTR           8948..9169
     misc_feature    8978
                     /label
                     /note="Geneious type: Editing History Insertion"
     primer_bind     complement(9015..9037)
                     /label="TK-SIN-3'UTR-seq-R"
     misc_feature    9169
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     complement(9169..9192)
                     /label="TK-SIN-3'UTR-R"
     CDS             9401..10258
                     /label="AmpR"
     promoter        11108..11125
                     /label="SP6"
ORIGIN      
        1 attgacggcg tagtacacac tattgaatca aacagccgac caattgcact accatcacaa
       61 tggagaagcc agtagtaaac gtagacgtag acccccagag tccgtttgtc gtgcaactgc
      121 aaaaaagctt cccgcaattt gaggtagtag cacagcaggt cactccaaat gaccatgcta
      181 atgccagagc attttcgcat ctggccagta aactaatcga gctggaggtt cctaccacag
      241 cgacgatctt ggacataggc agcgcaccgg ctcgtagaat gttttccgag caccagtatc
      301 attgtgtctg ccccatgcgt agtccagaag acccggaccg catgatgaaa tacgccagta
      361 aactggcgga aaaagcgtgc aagattacaa acaagaactt gcatgagaag attaaggatc
      421 tccggaccgt acttgatacg ccggatgctg aaacaccatc gctctgcttt cacaacgatg
      481 ttacctgcaa catgcgtgcc gaatattccg tcatgcagga cgtgtatatc aacgctcccg
      541 gaactatcta tcatcaggct atgaaaggcg tgcggaccct gtactggatt ggcttcgaca
      601 ccacccagtt catgttctcg gctatggcag gttcgtaccc tgcgtacaac accaactggg
      661 ccgacgagaa agtccttgaa gcgcgtaaca tcggactttg cagcacaaag ctgagtgaag
      721 gtaggacagg aaaattgtcg ataatgagga agaaggagtt gaagcccggg tcgcgggttt
      781 atttctccgt aggatcgaca ctttatccag aacacagagc cagcttgcag agctggcatc
      841 ttccatcggt gttccacttg aatggaaagc agtcgtacac ttgccgctgt gatacagtgg
      901 tgagttgcga aggctacgta gtgaagaaaa tcaccatcag tcccgggatc acgggagaaa
      961 ccgtgggata cgcggttaca cacaatagcg agggcttctt gctatgcaaa gttactgaca
     1021 cagtaaaagg agaacgggta tcgttccctg tgtgcacgta catcccggcc accatatgcg
     1081 atcagatgac tggtataatg gccacggata tatcacctga cgatgcacaa aaacttctgg
     1141 ttgggctcaa ccagcgaatt gtcattaacg gtaggactaa caggaacacc aacaccatgc
     1201 aaaattacct tctgccgatc atagcacaag ggttcagcaa atgggctaag gagcgcaagg
     1261 atgatcttga taacgagaaa atgctgggta ctagagaacg caagcttacg tatggctgct
     1321 tgtgggcgtt tcgcactaag aaagtacatt cgttttatcg cccacctgga acgcagacct
     1381 gcgtaaaagt cccagcctct tttagcgctt ttcccatgtc gtccgtatgg acgacctctt
     1441 tgcccatgtc gctgaggcag aaattgaaac tggcattgca accaaagaag gaggaaaaac
     1501 tgctgcaggt ctcggaggaa ttagtcatgg aggccaaggc tgcttttgag gatgctcagg
     1561 aggaagccag agcggagaag ctccgagaag cacttccacc attagtggca gacaaaggca
     1621 tcgaggcagc cgcagaagtt gtctgcgaag tggaggggct ccaggcggac atcggagcag
     1681 cattagttga aaccccgcgc ggtcacgtaa ggataatacc tcaagcaaat gaccgtatga
     1741 tcggacagta tatcgttgtc tcgccaaact ctgtgctgaa gaatgccaaa ctcgcaccag
     1801 cgcacccgct agcagatcag gttaagatca taacacactc cggaagatca ggaaggtacg
     1861 cggtcgaacc atacgacgct aaagtactga tgccagcagg aggtgccgta ccatggccag
     1921 aattcctagc actgagtgag agcgccacgt tagtgtacaa cgaaagagag tttgtgaacc
     1981 gcaaactata ccacattgcc atgcatggcc ccgccaagaa tacagaagag gagcagtaca
     2041 aggttacaaa ggcagagctt gcagaaacag agtacgtgtt tgacgtggac aagaagcgtt
     2101 gcgttaagaa ggaagaagcc tcaggtctgg tcctctcggg agaactgacc aaccctccct
     2161 atcatgagct agctctggag ggactgaaga cccgacctgc ggtcccgtac aaggtcgaaa
     2221 caataggagt gataggcaca ccggggtcgg gcaagtcagc tattatcaag tcaactgtca
     2281 cggcacgaga tcttgttacc agcggaaaga aagaaaattg tcgcgaaatt gaggccgacg
     2341 tgctaagact gaggggtatg cagattacgt cgaagacagt agattcggtt atgctcaacg
     2401 gatgccacaa agccgtagaa gtgctgtacg ttgacgaagc gttcgcgtgc cacgcaggag
     2461 cactacttgc cttgattgct atcgtcaggc cccgcaagaa ggtagtacta tgcggagacc
     2521 ccatgcaatg cggattcttc aacatgatgc aactaaaggt acatttcaat caccctgaaa
     2581 aagacatatg caccaagaca ttctacaagt atatctcccg gcgttgcaca cagccagtta
     2641 cagctattgt atcgacactg cattacgatg gaaagatgaa aaccacgaac ccgtgcaaga
     2701 agaacattga aatcgatatt acaggggcca caaagccgaa gccaggggat atcatcctga
     2761 catgtttccg cgggtgggtt aagcaattgc aaatcgacta tcccggacat gaagtaatga
     2821 cagccgcggc ctcacaaggg ctaaccagaa aaggagtgta tgccgtccgg caaaaagtca
     2881 atgaaaaccc actgtacgcg atcacatcag agcatgtgaa cgtgttgctc acccgcactg
     2941 aggacaggct agtgtggaaa accttgcagg gcgacccatg gattaagcag ctcactaaca
     3001 tacctaaagg aaactttcag gctactatag aggactggga agctgaacac aagggaataa
     3061 ttgctgcaat aaacagcccc actccccgtg ccaatccgtt cagctgcaag accaacgttt
     3121 gctgggcgaa agcattggaa ccgatactag ccacggccgg tatcgtactt accggttgcc
     3181 agtggagcga actgttccca cagtttgcgg atgacaaacc acattcggcc atttacgcct
     3241 tagacgtaat ttgcattaag tttttcggca tggacttgac aagcggactg ttttctaaac
     3301 agagcatccc actaacgtac catcccgccg attcagcgag gccggtagct cattgggaca
     3361 acagcccagg aacccgcaag tatgggtacg atcacgccat tgccgccgaa ctctcccgta
     3421 gatttccggt gttccagcta gctgggaagg gcacacaact tgatttgcag acggggagaa
     3481 ccagagttat ctctgcacag cataacctgg tcccggtgaa ccgcaatctt cctcacgcct
     3541 tagtccccga gtacaaggag aagcaacccg gcccggtcga aaaattcttg aaccagttca
     3601 aacaccactc agtacttgtg gtatcagagg aaaaaattga agctccccgt aagagaatcg
     3661 aatggatcgc cccgattggc atagccggtg cagataagaa ctacaacctg gctttcgggt
     3721 ttccgccgca ggcacggtac gacctggtgt tcatcaacat tggaactaaa tacagaaacc
     3781 accactttca gcagtgcgaa gaccatgcgg cgaccttaaa aaccctttcg cgttcggccc
     3841 tgaattgcct taactcagga ggcaccctcg tggtgaagtc ctatggctac gccgaccgca
     3901 acagtgagga cgtagtcacc gctcttgcca gaaagtttgt cagggtgtct gcagcgagac
     3961 cagattgtgt ctcaagcaat acagaaatgt acctgatttt ccgacaacta gacaacagcc
     4021 gtacacggca attcaccccg caccatctga attgcgtgat ttcgtccgtg tatgagggta
     4081 caagagatgg agttggagcc gcgccgtcat accgcaccaa aagggagaat attgctgact
     4141 gtcaagagga agcagttgtc aacgcagcca atccgctggg tagaccaggc gaaggagtct
     4201 gccgtgccat ctataaacgt tggccgacca gttttaccga ttcagccacg gagacaggca
     4261 ccgcaagaat gactgtgtgc ctaggaaaga aagtgatcca cgcggtcggc cctgatttcc
     4321 ggaagcaccc agaagcagaa gccttgaaat tgctacaaaa cgcctaccat gcagtggcag
     4381 acttagtaaa tgaacataac atcaagtctg tcgccattcc actgctatct acaggcattt
     4441 acgcagccgg aaaagaccgc cttgaagtat cacttaactg cttgacaacc gcgctagaca
     4501 gaactgacgc ggacgtaacc atctattgcc tggataagaa gtggaaggaa agaatcgacg
     4561 cggcactcca acttaaggag tctgtaacag agctgaagga tgaagatatg gagatcgacg
     4621 atgagttagt atggatccat ccagacagtt gcttgaaggg aagaaaggga ttcagtacta
     4681 caaaaggaaa attgtattcg tacttcgaag gcaccaaatt ccatcaagca gcaaaagaca
     4741 tggcggagat aaaggtcctg ttccctaatg accaggaaag taatgaacaa ctgtgtgcct
     4801 acatattggg tgagaccatg gaagcaatcc gcgaaaagtg cccggtcgac cataacccgt
     4861 cgtctagccc gcccaaaacg ttgccgtgcc tttgcatgta tgccatgacg ccagaaaggg
     4921 tccacagact tagaagcaat aacgtcaaag aagttacagt atgctcctcc accccccttc
     4981 ctaagcacaa aattaagaat gttcagaagg ttcagtgcac gaaagtagtc ctgtttaatc
     5041 cgcacactcc cgcattcgtt cccgcccgta agtacataga agtgccagaa cagcctaccg
     5101 ctcctcctgc acaggccgag gaggcccccg aagttgtagc gacaccgtca ccatctacag
     5161 ctgataacac ctcgcttgat gtcacagaca tctcactgga tatggatgac agtagcgaag
     5221 gctcactttt ttcgagcttt agcggatcgg acaactctat tactagtatg gacagttggt
     5281 cgtcaggacc tagttcacta gagatagtag accgaaggca ggtggtggtg gctgacgttc
     5341 atgccgtcca agagcctgcc cctattccac cgccaaggct aaagaagatg gcccgcctgg
     5401 cagcggcaag aaaagagccc actccaccgg caagcaatag ctctgagtcc ctccacctct
     5461 cttttggtgg ggtatccatg tccctcggat caattttcga cggagagacg gcccgccagg
     5521 cagcggtaca acccctggca acaggcccca cggatgtgcc tatgtctttc ggatcgtttt
     5581 ccgacggaga gattgatgag ctgagccgca gagtaactga gtccgaaccc gtcctgtttg
     5641 gatcatttga accgggcgaa gtgaactcaa ttatatcgtc ccgatcagcc gtatcttttc
     5701 cactacgcaa gcagagacgt agacgcagga gcaggaggac tgaatactga ctaaccgggg
     5761 taggtgggta catattttcg acggacacag gccctgggca cttgcaaaag aagtccgttc
     5821 tgcagaacca gcttacagaa ccgaccttgg agcgcaatgt cctggaaaga attcatgccc
     5881 cggtgctcga cacgtcgaaa gaggaacaac tcaaactcag gtaccagatg atgcccaccg
     5941 aagccaacaa aagtaggtac cagtctcgta aagtagaaaa tcagaaagcc ataaccactg
     6001 agcgactact gtcaggacta cgactgtata actctgccac agatcagcca gaatgctata
     6061 agatcaccta tccgaaacca ttgtactcca gtagcgtacc ggcgaactac tccgatccac
     6121 agttcgctgt agctgtctgt aacaactatc tgcatgagaa ctatccgaca gtagcatctt
     6181 atcagattac tgacgagtac gatgcttact tggatatggt agacgggaca gtcgcctgcc
     6241 tggatactgc aaccttctgc cccgctaagc ttagaagtta cccgaaaaaa catgagtata
     6301 gagccccgaa tatccgcagt gcggttccat cagcgatgca gaacacgcta caaaatgtgc
     6361 tcattgccgc aactaaaaga aattgcaacg tcacgcagat gcgtgaactg ccaacactgg
     6421 actcagcgac attcaatgtc gaatgctttc gaaaatatgc atgtaatgac gagtattggg
     6481 aggagttcgc tcggaagcca attaggatta ccactgagtt tgtcaccgca tatgtagcta
     6541 gactgaaagg ccctaaggcc gccgcactat ttgcaaagac gtataatttg gtcccattgc
     6601 aagaagtgcc tatggataga ttcgtcatgg acatgaaaag agacgtgaaa gttacaccag
     6661 gcacgaaaca cacagaagaa agaccgaaag tacaagtgat acaagccgca gaacccctgg
     6721 cgactgctta cttatgcggg attcaccggg aattagtgcg taggcttacg gccgtcttgc
     6781 ttccaaacat tcacacgctt tttgacatgt cggcggagga ttttgatgca atcatagcag
     6841 aacacttcaa gcaaggcgac ccggtactgg agacggatat cgcatcattc gacaaaagcc
     6901 aagacgacgc tatggcgtta accggtctga tgatcttgga ggacctgggt gtggatcaac
     6961 cactactcga cttgatcgag tgcgcctttg gagaaatatc atccacccat ctacctacgg
     7021 gtactcgttt taaattcggg gcgatgatga aatccggaat gttcctcaca ctttttgtca
     7081 acacagtttt gaatgtcgtt atcgccagca gagtactaga agagcggctt aaaacgtcca
     7141 gatgtgcagc gttcattggc gacgacaaca tcatacatgg agtagtatct gacaaagaaa
     7201 tggctgagag gtgcgccacc tggctcaaca tggaggttaa gatcatcgac gcagtcatcg
     7261 gtgagagacc accttacttc tgcggcggat ttatcttgca agattcggtt acttccacag
     7321 cgtgccgcgt ggcggatccc ctgaaaaggc tgtttaagtt gggtaaaccg ctcccagccg
     7381 acgacgagca agacgaagac agaagacgcg ctctgctaga tgaaacaaag gcgtggttta
     7441 gagtaggtat aacaggcact ttagcagtgg ccgtgacgac ccggtatgag gtagacaata
     7501 ttacacctgt cctactggca ttgagaactt ttgcccagag caaaagagca ttccaagcca
     7561 tcagagggga aataaagcat ctctacggtg gtcctaaata gtcagcatag tgccaccatg
     7621 gtgtctaagg gcgaagagct gattaaggag aacatgcaca tgaagctgta catggagggc
     7681 accgtgaaca accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
     7741 acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc
     7801 ctggctacca gcttcatgta cggcagcaaa accttcatca accacaccca gggcatcccc
     7861 gacttcttta agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa
     7921 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac
     7981 aacgtcaaga tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca
     8041 ctcggctggg aggcctccac cgagatgctg taccccgctg acggcggcct ggaaggcaga
     8101 agcgacatgg ccctgaagct cgtgggcggg ggccacctga tctgcaactt gaagaccaca
     8161 tacagatcca agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggacaga
     8221 agactggaaa gaatcaagga ggccgacaaa gagacctacg tcgagcagca cgaggtggct
     8281 gtggccagat actgcgacct ccctagcaaa ctggggcaca aacttaatgg aggcggaggc
     8341 ggaggcggag gcatgtacgt gagatttgag gttcctgagg acatgcagaa cgaagctctg
     8401 agtctgctgg agaaggttag ggagagcggt aaggtaaaga aaggtaccaa cgagacgaca
     8461 aaggctgtgg agaggggact ggcaaagctc gtttacatcg cagaggatgt tgacccgcct
     8521 gagatcgttg ctcatctgcc cctcctctgc gaggagaaga atgtgccgta catttacgtt
     8581 aaaagcaaga acgaccttgg aagggctgtg ggcattgagg tgccatgcgc ttcggcagcg
     8641 ataatcaacg agggagagct gagaaaggag cttggaagcc ttgtggagaa gattaaaggc
     8701 cttcagaaga agcttagcca tggcttcccg ccggaggtgg aggagcagga tgatggcacg
     8761 ctgcccatgt cttgtgccca ggagagcggg atggaccgtc accctgcagc ctgtgcttct
     8821 gctaggatca atgtgtaact cgacgcgccc cgcttgaagt ctttaattaa accgcttgaa
     8881 gtctttaatt aaaccgcttg aagtctttaa ttaaaccgct tgaagtcttt aattaaagct
     8941 agttaccctc gaggaagcgc agtacataat gctgcgcagt gttgccacat aaccactata
     9001 ttaaccattt atctagcgga cgccaaaaac tcaatgtatt tctgaggaag cgtggtgcat
     9061 aatgccacgc agcgtctgca taacttttat tatttctttt attaatcaac aaaattttgt
     9121 ttttaacatt tcaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaagg ggaattcgag
     9181 ctcgggaatt aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     9241 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     9301 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     9361 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     9421 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     9481 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     9541 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     9601 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     9661 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     9721 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     9781 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     9841 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     9901 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     9961 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
    10021 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
    10081 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
    10141 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
    10201 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
    10261 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
    10321 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
    10381 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
    10441 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
    10501 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
    10561 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
    10621 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
    10681 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
    10741 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
    10801 actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc
    10861 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
    10921 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
    10981 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcgagctc
    11041 gtatggacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac
    11101 acatacgatt taggtgacac tatag
//
