LOCUS       sequence-375199-       10745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector with altered Kozak sequence to direct 
            EGFP expression. mCherry expression is regulated by an IRES..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10745)
  AUTHORS   Ambrosini C, Destefanis E, Kheir E, Broso F, Alessandrini F, Longhi 
            S, Battisti N, Pesce I, Dassi E, Petris G, Cereseto A, Quattrone A
  TITLE     Translational enhancement by base editing of the Kozak sequence 
            rescues haploinsufficiency.
  JOURNAL   Nucleic Acids Res. 2022 Oct 14;50(18):10756-10771. doi: 
            10.1093/nar/gkac799.
  PUBMED    36165847
REFERENCE   2  (bases 1 to 10745)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2022-10-14- 14"; volume: "50"; issue: 
            "18"; pages: "10756-10771"
COMMENT     Sequence Label: pWPT-/mEGFP-1T-IRES-mCherry
FEATURES             Location/Qualifiers
     source          1..10745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             50..760
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(89..110)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(350..369)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     697..718
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(811..1385,1386..1388,1389..1397)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(978..995)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1205..1224
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      1406..1415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1412..2122
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1557..1575)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1825..1844
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2135..2723
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2188..2208)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2606..2617)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2868..2901)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             2921..3101
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(3126..3144)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3127..3144)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3434..3453)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3667..3686)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3786..3808
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3846..3864)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3932..4036
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4037..4105,4106..4897)
                     /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(4255..4274)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5068..5656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5557..5576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5810..5827
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5897..5917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5902..6231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6082..6217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          7365..7430
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7560..7580
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    8005..8139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8042..8061)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8096..8115
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             8307..8940
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    8987..9112
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9609..9842
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10027..10071
                     /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    10338..10455
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        10512..10723
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 ctaagcttac gctcgagtcc catctaacta agccactatg ggcgaattca gcaagggcga
       61 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      121 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      181 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
      241 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
      301 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
      361 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      421 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
      481 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
      541 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
      601 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
      661 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
      721 cgccgccggg atcactctcg gcatggacga gctgtacaag ttcgaataag catgcatcta
      781 gggcggccaa ttccgcccct ctcccccccc cccctctccc tccccccccc ctaacgttac
      841 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
      901 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
      961 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     1021 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     1081 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     1141 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     1201 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     1261 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     1321 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     1381 ataagcttgc cacaacccgg gatccgccac catggtgagc aagggcgagg aggataacat
     1441 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     1501 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     1561 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     1621 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     1681 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     1741 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     1801 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     1861 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     1921 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     1981 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     2041 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     2101 catggacgag ctgtacaagt aagcggccgc cgataatcaa cctctggatt acaaaatttg
     2161 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2221 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2281 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2341 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2401 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2461 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2521 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     2581 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2641 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2701 ctccctttgg gccgcctccc cgcatcgggc aattcgagct cggtaccttt aagaccaatg
     2761 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     2821 ctaattcact cccaacgaag acaagatggg atcaattcac catgggaata acttcgtata
     2881 gcatacatta tacgaagtta tgctgctttt tgcttgtact gggtctctct ggttagacca
     2941 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     3001 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     3061 atccctcaga cccttttagt cagtgtggaa aatctctagc agcatctaga attaattccg
     3121 tgtattctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat gtattaattg
     3181 tagccgcgtt ctaacgacaa tatgtacaag cctaattgtg tagcatctgg cttactgaag
     3241 cagaccctat catctctctc gtaaactgcc gtcagagtcg gtttggttgg acgaaccttc
     3301 tgagtttctg gtaacgccgt cccgcacccg gaaatggtca gcgaaccaat cagcagggtc
     3361 atcgctagcc agatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
     3421 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
     3481 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
     3541 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
     3601 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg aatggtgcac
     3661 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     3721 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3781 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     3841 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3901 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3961 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4021 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4081 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4141 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4201 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4261 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4321 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4381 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4441 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4501 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4561 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4621 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4681 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4741 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4801 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4861 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4921 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4981 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5041 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5101 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5161 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5221 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5281 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5341 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5401 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5461 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5521 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5581 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5641 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5701 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5761 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5821 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     5881 tcattaatgc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5941 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     6001 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     6061 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     6121 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     6181 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgga
     6241 cacaagacag gcttgcgaga tatgtttgag aataccactt tatcccgcgt cagggagagg
     6301 cagtgcgtaa aaagacgcgg actcatgtga aatactggtt tttagtgcgc cagatctcta
     6361 taatctcgcg caacctattt tcccctcgaa cactttttaa gccgtagata aacaggctgg
     6421 gacacttcac atgagcgaaa aatacatcgt cacctgggac atgttgcaga tccatgcacg
     6481 taaactcgca agccgactga tgccttctga acaatggaaa ggcattattg ccgtaagccg
     6541 tggcggtctg taccgggtgc gttactggcg cgtgaactgg gtattcgtca tgtcgatacc
     6601 gtttgtattt ccagctacga tcacgacaac cagcgcgagc ttaaagtgct gaaacgcgca
     6661 gaaggcgatg gcgaaggctt catcgttatt gatgacctgg tggataccgg tggtactgcg
     6721 gttgcgattc gtgaaatgta tccaaaagcg cactttgtca ccatcttcgc aaaaccggct
     6781 ggtcgtccgc tggttgatga ctatgttgtt gatatcccgc aagatacctg gattgaacag
     6841 ccgtgggata tgggcgtcgt attcgtcccg ccaatctccg gtcgctaatc ttttcaacgc
     6901 ctggcactgc cgggcgttgt tctttttaac ttcaggcggg ttacaatagt ttccagtaag
     6961 tattctggag gctgcatcca tgacacaggc aaacctgagc gaaaccctgt tcaaaccccg
     7021 ctttaaacat cctgaaacct cgacgctagt ccgccgcttt aatcacggcg cacaaccgcc
     7081 tgtgcagtcg gcccttgatg gtaaaaccat ccctcactgg tatcgcatga ttaaccgtct
     7141 gatgtggatc tggcgcggca ttgacccacg cgaaatcctc gacgtccagg cacgtattgt
     7201 gatgagcgat gccgaacgta ccgacgatga tttatacgat acggtgattg gctaccgtgg
     7261 cggcaactgg atttatgagt gggccccgga tctttgtgaa ggaaccttac ttctgtggtg
     7321 tgacataatt ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt
     7381 taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag attccaacct
     7441 atggaactga tgaatgggag cagtggtgga atgcctttaa tgaggaaaac ctgttttgct
     7501 cagaagaaat gccatctagt gatgatgagg ctactgctga ctctcaacat tctactcctc
     7561 caaaaaagaa gagaaaggta gaagacccca aggactttcc ttcagaattg ctaagttttt
     7621 tgagtcatgc tgtgtttagt aatagaactc ttgcttgctt tgctatttac accacaaagg
     7681 aaaaagctgc actgctatac aagaaaatta tggaaaaata ttctgtaacc tttataagta
     7741 ggcataacag ttataatcat aacatactgt tttttcttac tccacacagg catagagtgt
     7801 ctgctattaa taactatgct caaaaattgt gtacctttag ctttttaatt tgtaaagggg
     7861 ttaataagga atatttgatg tatagtgcct tgactagaga tcataatcag ccataccaca
     7921 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     7981 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     8041 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     8101 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
     8161 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
     8221 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
     8281 taaatatgta ctacaaactt agtagttgga agggctaatt cactcccaaa gaagacaaga
     8341 tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc agaactacac
     8401 accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     8461 tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt
     8521 gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg
     8581 cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata
     8641 tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg
     8701 gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg
     8761 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     8821 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     8881 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     8941 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     9001 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     9061 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     9121 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     9181 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     9241 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     9301 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     9361 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     9421 aaacaaaagt aagaccaccg cacagcaagc ggccggccgc tgatcttcag acctggagga
     9481 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     9541 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     9601 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     9661 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     9721 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     9781 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     9841 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     9901 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     9961 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
    10021 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
    10081 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
    10141 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
    10201 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
    10261 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
    10321 cgacggtatc ggttaacttt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
    10381 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
    10441 aaaattcaaa attttatcga tcacgagact agcctcgaca agcttgatcg atggctccgg
    10501 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
    10561 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
    10621 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
    10681 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtgtcgt gacgcgcatc
    10741 caggc
//
