LOCUS       sequence-434956-       10099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses VHL-BirA for VHL substrate identification by E-STUB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10099)
  AUTHORS   Huang HT, Lumpkin RJ, Tsai RW, Su S, Zhao X, Xiong Y, Chen J, Mageed
            N, Donovan KA, Fischer ES, Sellers WR
  TITLE     Ubiquitin-specific proximity labeling for the identification of E3 
            ligase substrates.
  JOURNAL   Nat Chem Biol. 2024 Mar 21. doi: 10.1038/s41589-024-01590-9.
  PUBMED    38514884
REFERENCE   2  (bases 1 to 10099)
  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: "Nat Chem 
            Biol. 2024 Mar 21. doi: 10.1038/s41589-024-01590-9."
COMMENT     Sequence Label: pLX307-VHL-3F-BirA
FEATURES             Location/Qualifiers
     source          1..10099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..1274
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          327..1265
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1222..1242
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1294..1318
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1972..2037
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2041..3006
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             2041..3003
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     protein_bind    complement(3008..3032)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3040..3081
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3132..3720
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3185..3205)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3603..3614)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3792..4025
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4103..4224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4140..4159)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4194..4213
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4264..4399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4419..4438)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4420..4438)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4448..4465)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4448..4464)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4457..4479)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4606..5061
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4693..4712)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4903..4924
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5087..5191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5192..5260,5261..6052)
                     /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(5410..5429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6223..6811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6712..6731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6965..6982
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7099..7120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7135..7140,7141..7158,7159..7165)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7173..7189
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7178..7200
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7197..7213
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7197..7213
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7232..7252
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7234..7252
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7280..7506
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7507..7687
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7734..7859
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8352..8585
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8770..8814
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        9045..9255
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        9067..9263
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      9114..9249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             9295..9897
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
     primer_bind     complement(9295..9314)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9791..9811
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9940..10057
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ctcgaggcct gcaggtgcaa agatggataa agttttaaac agagaggaat ctttgcagct
       61 aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
      121 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
      181 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      241 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      301 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
      361 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga
      421 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
      481 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
      541 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
      601 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
      661 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
      721 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
      781 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
      841 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
      901 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
      961 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     1021 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt
     1081 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     1141 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     1201 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     1261 ttcaggtgtc gtgaggctag catcgattga tcaacaagtt tgtacaaaaa agcaggctta
     1321 gccaccatgc cacggagagc cgagaactgg gatgaggccg aggtgggcgc tgaagaggcc
     1381 ggcgtggaag agtacggccc cgaggaagat ggcggagaag aaagcggcgc cgaggagagc
     1441 ggacctgagg agagcggccc tgaggaactg ggagccgagg aagaaatgga agccggaaga
     1501 cctagacccg tgcttagaag cgtgaacagc agagagccat ctcaggtgat tttctgcaac
     1561 agatccccta gagtggtgct gcctgtgtgg ctgaacttcg acggcgagcc tcagccctac
     1621 cccaccctgc cccccggcac cggcagacgg atccattcct atagaggcca cctgtggctg
     1681 tttcgggacg ccggcaccca cgacggcctg ctggttaacc agaccgagct gttcgtgcct
     1741 tctctgaatg tggacggcca gcctatcttc gccaacatca ccctgcctgt gtacacactg
     1801 aaggaaaggt gtctgcaggt cgtgcggagc ctggtcaagc ctgaaaatta ccggaggctg
     1861 gacatcgtgc ggagcctgta cgaggatctg gaagatcacc ctaacgtgca gaaagacctc
     1921 gagcgcctga cacaggagag aatcgctcac caaagaatgg gcgacggttc tgactacaaa
     1981 gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga tgacaagctt
     2041 atgaaggata acaccgtgcc actgaaattg attgccctgt tagcgaacgg tgaatttcac
     2101 tctggcgagc agttgggtga aacgctggga atgagccggg cggctattaa taaacacatt
     2161 cagacactgc gtgactgggg cgttgatgtc tttaccgttc cgggtaaagg atacagcctg
     2221 cctgagccta tccagttact taatgctaaa cagatattgg gtcagctgga tggcggtagt
     2281 gtagccgtgc tgccagtgat tgactccacg aatcagtacc ttcttgatcg tatcggagag
     2341 cttaaatcgg gcgatgcttg cattgcagaa taccagcagg ctggccgtgg tcgccggggt
     2401 cggaaatggt tttcgccttt tggcgcaaac ttatatttgt cgatgttctg gcgtctggaa
     2461 caaggcccgg cggcggcgat tggtttaagt ctggttatcg gtatcgtgat ggcggaagta
     2521 ttacgcaagc tgggtgcaga taaagttcgt gttaaatggc ctaatgacct ctatctgcag
     2581 gatcgcaagc tggcaggcat tctggtggag ctgactggca aaactggcga tgcggcgcaa
     2641 atagtcattg gagccgggat caacatggca atgcgccgtg ttgaagagag tgtcgttaat
     2701 caggggtgga tcacgctgca ggaagcgggg atcaatctcg atcgtaatac gttggcggcc
     2761 atgctaatac gtgaattacg tgctgcgttg gaactcttcg aacaagaagg attggcacct
     2821 tatctgtcgc gctgggaaaa gctggataat tttattaatc gcccagtgaa acttatcatt
     2881 ggtgataaag aaatatttgg catttcacgc ggaatagaca aacagggggc tttattactt
     2941 gagcaggatg gaataataaa accctggatg ggcggtgaaa tatccctgcg tagtgcagaa
     3001 aaatgaaacc cagctttctt gtacaaagtg gttgatatcg gtaagcctat ccctaaccct
     3061 ctcctcggtc tcgattctac gtagtaatga actagtaccg gttaagtcga caatcaacgc
     3121 gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3181 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3241 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3301 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3361 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3421 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3481 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     3541 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3601 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3661 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3721 gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     3781 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     3841 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     3901 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     3961 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     4021 tagcagtacg tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     4081 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     4141 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4201 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     4261 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4321 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4381 ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg
     4441 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     4501 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     4561 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     4621 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4681 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4741 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4801 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4861 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4921 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4981 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5041 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5101 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5161 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5221 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5281 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5341 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5401 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5461 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5521 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5581 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5641 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5701 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5761 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5821 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5881 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5941 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6001 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6061 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6121 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6181 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6241 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6301 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6361 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6841 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6901 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6961 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     7021 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     7081 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     7141 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     7201 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
     7261 gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta
     7321 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
     7381 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
     7441 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
     7501 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     7561 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     7621 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     7681 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     7741 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     7801 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     7861 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     7921 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     7981 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     8041 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     8101 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     8161 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     8221 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     8281 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     8341 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     8401 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     8461 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     8521 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     8581 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     8641 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     8701 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     8761 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     8821 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     8881 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     8941 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccat gcattgcatc
     9001 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     9061 aaagcatgcg tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     9121 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     9181 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     9241 ttggaggcct aggcttttgc aaaaagcttt ctagaggtac caccggagct taccatgacc
     9301 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     9361 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     9421 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     9481 aaggtgtggg tcgcggacga cggcgccgcc gtggcggtct ggaccacgcc ggagagcgtc
     9541 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     9601 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     9661 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     9721 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     9781 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     9841 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc cagttaacga
     9901 attctagatc ttgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     9961 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
    10021 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
    10081 agatccactt tggcgccgg
//
