pLVX-puro-cOVA-IRES-BFP

LOCUS        pLVX-puro-cOVA-IRES-BFP       10692 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses chicken cytoplasmic ovalbumin in lentiviral vector with 
            puromycin resistance. cOVA is coupled with BFP fluorescent reporter 
            using IRES..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10692)
  TITLE     pLVX-OVA
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10692)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pLVX-puro-cOVA-IRES-BFP
FEATURES             Location/Qualifiers
     source          1..10692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    47..596
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(191..208)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     418..437
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(599..601,602..604,605..1318)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(644..665)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    1440..1561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1614..2113
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1649..1672)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2046..2065
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2134..2733
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2134..2153)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2630..2650
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2747..3335
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2800..2820)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3218..3229)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3542..4175
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4304..4320)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4304..4320)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4317..4339)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4328..4344
                     /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)."
     promoter        complement(join(4352..4358,4359..4376,4377..4382))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4397..4418
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4535..4552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4706..5294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4786..4805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5465..6256,6257..6325))
                     /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     6088..6107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6326..6430)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6478..6612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             6780..7413
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7460..7585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8082..8315
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8500..8544
                     /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    8807..8922
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     enhancer        8980..9283
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9284..9495
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9446..9466
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9492..9516
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          9656..9788
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     9771..9790
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        9833..9851
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             10267..10290
                     /codon_start=1
                     /product="antigenic CD8+ T cell epitope from chicken 
                     ovalbumin"
                     /label=OVA peptide
                     /translation="SIINFEKL"
ORIGIN
        1 ccgggatccc aattgctcga gcgggatcaa ttccgccccc cccctaacgt tactggccga
       61 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
      121 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
      181 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
      241 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
      301 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
      361 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
      421 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
      481 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
      541 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg ataataccat
      601 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      661 cgacgtaaac ggccacaagt tcagcgtgag gggcgagggc gagggcgatg ccaccaacgg
      721 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      781 cgtgaccacc ctgagccacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
      841 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      901 caaggacgac ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      961 gaaccgcatc gagctgaagg gcgtcgactt caaggaggac ggcaacatcc tggggcacaa
     1021 gctggagtac aacttcaaca gccacaacat ctatatcatg gccgtcaagc agaagaacgg
     1081 catcaaggtg aacttcaaga tccgccacaa cgtggaggac ggcagcgtgc agctcgccga
     1141 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acagccacta
     1201 cctgagcacc cagtccgtgc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1261 gctggagttc cgcaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagc
     1321 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     1381 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     1441 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1501 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1561 aaggcgtaaa ttgtaagcgt taatgggatc ccgcgactct agataattct accgggtagg
     1621 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg
     1681 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc
     1741 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt
     1801 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac
     1861 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag
     1921 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc
     1981 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg
     2041 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc
     2101 tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc
     2161 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     2221 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     2281 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     2341 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     2401 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     2461 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     2521 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     2581 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     2641 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     2701 acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc tggaacaatc aacctctgga
     2761 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2821 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2881 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2941 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3001 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3061 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3121 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3181 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3241 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3301 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac
     3361 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
     3421 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
     3481 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga
     3541 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac
     3601 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca
     3661 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc
     3721 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg
     3781 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga
     3841 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc
     3901 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga
     3961 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
     4021 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     4081 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     4141 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     4201 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta
     4261 gcgttttacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4321 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4381 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4441 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4501 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4561 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4621 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5581 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5761 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6061 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6361 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6421 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac
     6481 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6541 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6601 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
     6661 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
     6721 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt
     6781 ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga tctaccacac
     6841 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
     6901 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
     6961 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
     7021 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
     7081 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
     7141 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
     7201 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
     7261 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     7321 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     7381 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     7441 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     7501 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     7561 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     7621 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     7681 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     7741 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     7801 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     7861 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     7921 agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     7981 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     8041 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     8101 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     8161 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     8221 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     8281 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     8341 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     8401 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     8461 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     8521 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     8581 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     8641 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     8701 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     8761 agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccttta aaagaaaagg
     8821 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     8881 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     8941 cagcagagat ccagtttatc gataagcttg ggagttccgc gttacataac ttacggtaaa
     9001 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     9061 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     9121 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt
     9181 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc
     9241 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     9301 gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     9361 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     9421 caactgcgat cgcccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     9481 tatataagca gagctcgttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt
     9541 tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc
     9601 tgcagtgact ctcttaaggt agccttgcag aagttggtcg tgaggcactg ggcaggtaag
     9661 tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag
     9721 aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct
     9781 ctccacaggt gtccactccc agttcaatta cagctcttaa ggctagagta cttaatacga
     9841 ctcactatag gctagcctcg agaattcgtc gacatgggct ccatcggcgc agcaagcatg
     9901 gaattttgtt ttgatgtatt caaggagctc atcaattcct gggtagaaag tcagacaaat
     9961 ggaattatca gaaatgtcct tcagccaagc tccgtggatt ctcaaactgc aatggttctg
    10021 gttaatgcca ttgtcttcaa aggactgtgg gagaaaacat ttaaggatga agacacacaa
    10081 gcaatgcctt tcagagtgac tgagcaagaa agcaaacctg tgcagatgat gtaccagatt
    10141 ggtttattta gagtggcatc aatggcttct gagaaaatga agatcctgga gcttccattt
    10201 gccagtggga caatgagcat gttggtgctg ttgcctgatg aagtctcagg ccttgagcag
    10261 cttgagagta taatcaactt tgaaaaactg actgaatgga ccagttctaa tgttatggaa
    10321 gagaggaaga tcaaagtgta cttacctcgc atgaagatgg aggaaaaata caacctcaca
    10381 tctgtcttaa tggctatggg cattactgac gtgtttagct cttcagccaa tctgtctggc
    10441 atctcctcag cagagagcct gaagatatct caagctgtcc atgcagcaca tgcagaaatc
    10501 aatgaagcag gcagagaggt ggtagggtca gcagaggctg gagtggatgc tgcaagcgtc
    10561 tctgaagaat ttagggctga ccatccattc ctcttctgta tcaagcacat cgcaaccaac
    10621 gccgttctct tctttggcag atgtgtttcc ccttaagtcg acgaattcac gcgtggtacc
    10681 tctagagtcg ac
//