LOCUS       sequence-459009-       10511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Validates the expression and detection of epitope-tagged proteins of
            interest in Drosophila cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10511)
  AUTHORS   Robinson E, Alonso EB, A Waters J, Bileckyj C, D House C, A Johnston
            C, M Cripps R
  TITLE     pJoseph2: a family of plasmids as positive controls for bacterial 
            protein expression, transfections, and western blots.
  JOURNAL   Biotechniques. 2024;76(7):299-309. doi: 
            10.1080/07366205.2024.2343609. Epub 2024 May 12.
  PUBMED    39185782
REFERENCE   2  (bases 1 to 10511)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Biotechniques."; date: "2024"; volume: "76(7):299-309. doi"; pages:
            " 10.1080/07366205.2024.2343609. Epub 2024 May 12"
COMMENT     Sequence Label: pAHW/Joseph2-Multitagged Plasmid
FEATURES             Location/Qualifiers
     source          1..10511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             600..671
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             702..743
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             744..785
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             786..827
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             858..875
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(880..904)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          996..1061
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1191..1211
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1636..1770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1673..1692)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1727..1746
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             join(1867..1935,1936..2727)
                     /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     complement(2085..2104)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2898..3486
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3808..6317
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     regulatory      6356..6365
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6395..6484
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     protein_bind    6506..6530
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6545..6634
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             6665..7378
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
                     NFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILLE
                     FVTAAGITHGMDELYK"
     CDS             6665..7378
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
                     NFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILLE
                     FVTAAGITHGMDELYK"
     CDS             7409..7480
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             7511..7552
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7553..7594
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7595..7636
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7667..7684
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(7689..7788)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7805..7824)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7806..7824)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7829..7845)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7829..7845)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7947..7966)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7958..8767
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(8025..8044)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      8914..9502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(9832..9859)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(9951..10037)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     10086..10108
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10100..10117
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10101..10117
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    10133..10232
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             10247..10336
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             join(10367..10511,1..569)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
                     NFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILLE
                     FVTAAGITHGMDELYK"
     CDS             join(10367..10511,1..569)
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
                     NFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILLE
                     FVTAAGITHGMDELYK"
ORIGIN
        1 ctactgggaa gctacctgtt ccatggccaa cacttgtcac tactttgact tatggtgttc
       61 aatgcttttc aagataccca gatcatatga aacagcatga ctttttcaag agtgccatgc
      121 ccgaaggtta tgtacaggaa agaactatat tttacaaaga tgacgggaac tacaagacac
      181 gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg
      241 attttaaaga agatggaaac attcttggac acaaaatgga atacaactat aactcacata
      301 atgtatacat catggcagac aaaccaaaga atggaatcaa agttaacttc aaaattagac
      361 acaacattaa agatggaagc gttcaattag cagaccatta tcaacaaaat actccaattg
      421 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcca
      481 aagatcccaa cgaaaagaga gatcacatga tccttcttga gtttgtaaca gctgctggga
      541 ttacacatgg catggatgaa ctatacaaag gtagtggttc cggtagcggc agtgggagtg
      601 attataaaga cgatgatgac aaggactata aggacgacga cgacaaggat tacaaggatg
      661 atgacgacaa aggctccggt tcggggtcgg gaagcggtag tggtaaaccg attcccaatc
      721 cccttcttgg tcttgattct acaggcaaac ccatcccgaa cccactcctc ggccttgact
      781 ccacaggcaa gcctattcca aatcctctac taggcctcga ttctacgggt agcggttcgg
      841 gatccggaag tggaagccac catcatcatc accattagga cccagctttc ttgtacaaag
      901 tggtgacgta agctagcagg atctttgtga aggaacctta cttctgtggt gtgacataat
      961 tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat
     1021 aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc tatggaactg
     1081 atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc tcagaagaaa
     1141 tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct ccaaaaaaga
     1201 agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt ttgagtcatg
     1261 ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag gaaaaagctg
     1321 cactgctata caagaaaatt atggaaaaat attctgtaac ctttataagt aggcataaca
     1381 gttataatca taacatactg ttttttctta ctccacacag gcatagagtg tctgctatta
     1441 ataactatgc tcaaaaattg tgtaccttta gctttttaat ttgtaaaggg gttaataagg
     1501 aatatttgat gtatagtgcc ttgactagag atcataatca gccataccac atttgtagag
     1561 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     1621 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1681 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1741 ctcatcaatg tatcttatca tgtctggatc ccgtttaaac tacgcgtaat tcaaacaggg
     1801 ttctggcgtc gttctcgtac tgttttcccc aggccagtgc tttagcgtta ttgaaaaagg
     1861 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1921 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1981 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     2041 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     2101 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     2161 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     2221 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     2281 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2341 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2401 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2461 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2521 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2581 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2641 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2701 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2761 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2821 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2881 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2941 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3001 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3061 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3121 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3181 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3241 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3301 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3361 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3421 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3481 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3541 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3601 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3661 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3721 aggcaactcg tgaaaggtag gcggatcagc ggccgcagca tgcaattcta tattctaaaa
     3781 acacaaatga tacttctaaa aaaaaatcat gaatggcatc aactctgaat caaatctttg
     3841 cagatgcacc tacttctcat ttccactgtc acatcatttt tccagatctc gctgcctgtt
     3901 atgtggccca caaaccaaga cacgttttat ggccattaaa gctggctgat cgtcgccaaa
     3961 caccaaatac ataatgaata tgtacacatt cgagaaagaa gcgatcaaag aagcgtcttc
     4021 gggcggagta ggagaatgcg gaggagaagg agaacgagct gatctagtat ctctccacaa
     4081 tccaatgcca actgaccaac tggccatatt cggagcaatt tgaagccaat ttccatcgcc
     4141 tggcgatcgc tccattcttg gctatatgtt tttcaccgtt acccggggcc attttcaaag
     4201 actcgtcggc aagataagat tgtgtcactc gctgtctctc ttcatttgtc gaagaatgct
     4261 gaggaatttc gcgatgacgt cggcgagtat tttgaagaat gagaataatt tgtatttata
     4321 cgaaaatcag ttagtggaat tttctacaaa aacatgttat ctatagataa ttttgttgca
     4381 aaatatgttg actatgacaa agattgtatg tatatacctt taatgtattc tcattttctt
     4441 atgtatttat aatggcaatg atgatactga tgatatttta agatgatgcc agaccaaaag
     4501 gcttgaattt ctgcgtcttt tgccgaacgc agtgcatgtg caattgttgt tttttggaat
     4561 attcaatttt cggactgtcc gctttgattt cagtttcttg gcttattcaa aaagcaaagt
     4621 aaagccaaaa aagcgagatg gcaataccaa atgcggcaaa acggtagtgg aaggaaaggg
     4681 gtgcggggca gcggaaggaa gggtggggcg gggcgtggcg gggtctgtgg ctgggcgcga
     4741 cgtcaccgac gttggagcca ctcctttgac catgtgtgcg tgtgtgtatt attcgtgtct
     4801 cgccactcgc cggttgtttt tttcttttta tgctgcgctc tctctagcgc catctcgctt
     4861 acgcatgctc aacgcaccgc atgttgccgt ttccttttat gcgtcatttt ggctcgaaat
     4921 aggcaattat ttaaacaaag attagtcaac gaaaacgcta aaataaataa gtctacaata
     4981 tggttactta ttgccatgtg tgtgcagcca acgatagcaa caaaagcaac aacacaggtg
     5041 gctttccctc tttcactttt tgtttgcaag ccgcgtgcga gcaagacggc acgaccggca
     5101 aacgcaatta cgctgacaaa gagcagacga agttttggcg aaaaacatca aggcgcctga
     5161 tacgaatgca tttgcaataa caattgcgat atttaatatt gtttatgaag ctgtttgact
     5221 tcaaaacaca caaaaaaaaa aataaaacaa attatttgaa agagaattag gaatcggacg
     5281 cttatcgtta gggtaacaac aagaaatgct tactgagtca cagcctctgg aaaactgccg
     5341 caagccagag agagagagaa aaagagggag agcagcttag accgcatgtg cttgtgtgtg
     5401 aggcgtctct ctcttcgtct ctgttgcgca aacgcataga ctgcactgaa aaaatcgatt
     5461 acctattttt tatgaatgaa tatttgcact attactattc aaaactatta agatagcaat
     5521 cacattcaat agccaaatac tataccacct gagcgatgca acgaaatgat caatttgagc
     5581 aaaaatgctg catatttagg acggcatcat tatagaaatg cttcttgctg tgtacttttc
     5641 tctcgtctgg cagctgtttc gccgttattg ttaaaaccgg cttaagttag gtgtgttttc
     5701 tacgactagt gaatgcccta ctagaagatg tgtgttgcac aaaatgtccc tggaataacc
     5761 aatttgaagt gcagatagca gtaaacgtaa gctaatatga atattattta actgtaatgt
     5821 tttaatatcg ctggacatta ctaataaacc cactataaac acatgtacat atgtatgttt
     5881 tggcatacaa tgagtagttg gggaaaaaat gtgtaaaagc accgtgacca tcacagcata
     5941 aagataacca gctgaagtat cgaatatgag taacccccaa attgaatcac atgccgcaac
     6001 tgataggacc catggaagta cactcttcat ggcgatatac aagacacaca caagcacgaa
     6061 cacccagttg cggaggaaat tctccgtaaa tgaaaaccca atcggcgaac aattcatacc
     6121 catatatggt aaaagttttg aacgcgactt gagagcggag agcattgcgg ctgataaggt
     6181 tttagcgcta agcgggcttt ataaaacggg ctgcgggacc agttttcata tcactaccgt
     6241 ttgagttctt gtgctgtgtg gatactcctc ccgacacaaa gccgctccat cagccagcag
     6301 tcgtctaatc cagagacccc ggatccagat atcgaggcct gtctagagaa gctccgccac
     6361 catggatctc caccgcggtg gaggccgcat cttttaccca tacgatgttc ctgactatgc
     6421 gggctatccc tatgacgtcc cggactatgc aggatcctat ccatatgacg ttccagatta
     6481 cgctgctcat ggcggacacc ggtatacaag tttgtacaaa aaagcaggct tcggcagcgg
     6541 aagcgagcaa aagctcatct cggaggaaga tctagaacag aagctaatat cagaggagga
     6601 cctagagcaa aaacttattt cggaagaaga tctgggcagc ggatccggtt cgggaagcgg
     6661 tagtatgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
     6721 tggcgatgtt aatgggcaaa aattctctgt cagtggagag ggtgaaggtg atgcaacata
     6781 cggaaaactt acccttaaat ttatttgcac tactgggaag ctacctgttc catggccaac
     6841 acttgtcact actttgactt atggtgttca atgcttttca agatacccag atcatatgaa
     6901 acagcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt
     6961 ttacaaagat gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct
     7021 tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca
     7081 caaaatggaa tacaactata actcacataa tgtatacatc atggcagaca aaccaaagaa
     7141 tggaatcaaa gttaacttca aaattagaca caacattaaa gatggaagcg ttcaattagc
     7201 agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca
     7261 ttacctgtcc acacaatctg ccctttccaa agatcccaac gaaaagagag atcacatgat
     7321 ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaagg
     7381 tagtggttcc ggtagcggca gtgggagtga ttataaagac gatgatgaca aggactataa
     7441 ggacgacgac gacaaggatt acaaggatga tgacgacaaa ggctccggtt cggggtcggg
     7501 aagcggtagt ggtaaaccga ttcccaatcc ccttcttggt cttgattcta caggcaaacc
     7561 catcccgaac ccactcctcg gccttgactc cacaggcaag cctattccaa atcctctact
     7621 aggcctcgat tctacgggta gcggttcggg atccggaagt ggaagccacc atcatcatca
     7681 ccattaggac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     7741 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     7801 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     7861 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     7921 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     7981 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     8041 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     8101 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     8161 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     8221 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     8281 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     8341 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     8401 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     8461 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     8521 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     8581 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     8641 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     8701 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     8761 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     8821 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     8881 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     8941 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     9001 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     9061 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     9121 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     9181 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     9241 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     9301 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     9361 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     9421 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     9481 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     9541 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     9601 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     9661 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     9721 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     9781 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
     9841 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
     9901 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
     9961 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
    10021 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
    10081 gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat
    10141 gattttattt tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt
    10201 tataatgcca actttgtaca aaaaagcagg cttcggcagc ggaagcgagc aaaagctcat
    10261 ctcggaggaa gatctagaac agaagctaat atcagaggag gacctagagc aaaaacttat
    10321 ttcggaagaa gatctgggca gcggatccgg ttcgggaagc ggtagtatga gtaaaggaga
    10381 agaacttttc actggagttg tcccaattct tgttgaatta gatggcgatg ttaatgggca
    10441 aaaattctct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa
    10501 atttatttgc a
//
