LOCUS       sequence-472682-       10562 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Expression of structurally predicted WWE domain of HUWE1 with EGFP 
            on C terminal .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10562)
  TITLE     Sobol Lab WWE Plasmids 2025
REFERENCE   2  (bases 1 to 10562)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pLV-EF1A-HUWE1-1503-1780_EGFP-Hygro
FEATURES             Location/Qualifiers
     source          1..10562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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"
     misc_feature    1803..1920
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    1938..1958
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1959..3137
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2190..3128
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3085..3105
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    3137..3161
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3162..3171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4089..4091,4092..4094,4095..4808)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4089..4091,4092..4094,4095..4805)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4134..4155)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4395..4414)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4742..4763
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4809..4833)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4851..5439
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4904..4924)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5322..5333)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        5470..5969
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5505..5528)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5902..5921
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5986..7011
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             5986..7011
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             7083..7316
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7388..7509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7425..7444)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7479..7498
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7549..7684
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7704..7723)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7705..7723)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7733..7750)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7733..7749)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7742..7764)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7891..8346
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7978..7997)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8188..8209
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8372..8476
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8477..8545,8546..9337)
                     /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(8695..8714)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9508..10096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9997..10016
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10250..10267
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10384..10405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10420..10425,10426..10443,10444..10450)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10458..10474
                     /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     10463..10485
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10482..10498
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10482..10498
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10517..10537
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10519..10537
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcta
     1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     1921 actagtgatt atcggatcaa ctttgtatag aaaagttggg ctccggtgcc cgtcagtggg
     1981 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg
     2041 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     2101 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     2161 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg
     2221 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg
     2281 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
     2341 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
     2401 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
     2461 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
     2521 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
     2581 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
     2641 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg tctcgcgccg ccgtgtatcg
     2701 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
     2761 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
     2821 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
     2881 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     2941 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     3001 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     3061 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     3121 catttcaggt gtcgtgacaa gtttgtacaa aaaagcaggc tgccaccatg gccgccgacg
     3181 tgctgatcaa ggccgccctg cccctgacta ccagcgacac taagacagtg tccgaatgga
     3241 tttcccagat ggctactctg ccacaggcca gcaatctggc cacccgcatc ctgctgctga
     3301 ccctgctgtt cgaagagctg aagctgccct gcgcctgggt ggtggagtcc agcgggatcc
     3361 tgaacgtgct gattaagctg cttgaggtgg tgcagccatg cctgcaggct gccaaggaac
     3421 agaaagaggt gcagacccca aagtggatta cccccgttct gctgctgatc gacttttatg
     3481 aaaagacagc tatctcttcc aagagacggg cacagatgac caagtacctg cagtcaaact
     3541 ccaacaattg gcgctggttc gatgaccgat ctgggaggtg gtgctcttat agcgcctcta
     3601 ataattccac tatcgactcc gcctggaaga gcggcgagac ctctgtgcgc ttcaccgccg
     3661 gcaggagaag gtacacagtg cagtttacta ccatggtgca ggtgaacgag gaaacaggca
     3721 atcggcggcc cgtgatgctg accctcctgc gcgtgccccg cctgaacaag aatagcaaga
     3781 acagcaacgg gcaggagctg gagaagaccc tggaggagtc caaagagatg gatatcaaga
     3841 gaaaggaaaa caagggcaat gacacccccc tggccctgga gtctaccaac accgagaagg
     3901 agacctctct ggaggagacc aaaatcggcg agatcctgat ccagggcctg accgaggaca
     3961 tggtgactgt cctgatccgg gcttgcgtgt ctatgctggg cgtgcctgtg gaccccgaca
     4021 ccctgcacgc caccctgcgg tccggcggcg gcggctccgg aggcggcggc tccgggggcg
     4081 gcggctccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     4141 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     4201 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     4261 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     4321 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     4381 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     4441 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     4501 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     4561 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     4621 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     4681 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     4741 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     4801 acaagtaaac ccagctttct tgtacaaagt ggtgataatc gaattccgat aatcaacctc
     4861 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     4921 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     4981 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     5041 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     5101 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     5161 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     5221 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     5281 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     5341 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     5401 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgggaattc ccgcggttcg
     5461 aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
     5521 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
     5581 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
     5641 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
     5701 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
     5761 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
     5821 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc
     5881 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
     5941 tctcctcttc ctcatctccg ggcctttcga cctcacgtgg ccaccatgaa aaagcctgaa
     6001 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg
     6061 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga
     6121 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg
     6181 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga atttagcgag
     6241 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa
     6301 accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc
     6361 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
     6421 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
     6481 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
     6541 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg
     6601 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
     6661 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
     6721 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat
     6781 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
     6841 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg
     6901 cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc
     6961 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata gggtaccttt
     7021 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     7081 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
     7141 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     7201 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     7261 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     7321 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     7381 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     7441 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     7501 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta
     7561 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     7621 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     7681 ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     7741 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     7801 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     7861 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     7921 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     7981 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     8041 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     8101 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     8161 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     8221 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     8281 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     8341 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     8401 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     8461 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     8521 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     8581 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     8641 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     8701 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     8761 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     8821 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     8881 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     8941 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     9001 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     9061 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     9121 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     9181 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     9241 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     9301 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     9361 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     9421 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     9481 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     9541 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     9601 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     9661 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     9721 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     9781 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     9841 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     9901 atgagaaagc gccacgcttc ccgaagagag aaaggcggac aggtatccgg taagcggcag
     9961 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
    10021 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
    10081 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
    10141 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
    10201 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
    10261 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
    10321 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
    10381 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
    10441 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
    10501 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
    10561 tt
//
