LOCUS       sequence-392111-       13644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct to stably express NPC1 TurboID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13644)
  AUTHORS   Shin HR, Citron YR, Wang L, Tribouillard L, Goul CS, Stipp R, 
            Sugasawa Y, Jain A, Samson N, Lim CY, Davis OB, Castaneda-Carpio D, 
            Qian M, Nomura DK, Perera RM, Park E, Covey DF, Laplante M, Evers 
            AS, Zoncu R
  TITLE     Lysosomal GPCR-like protein LYCHOS signals cholesterol sufficiency 
            to mTORC1.
  JOURNAL   Science. 2022 Sep 16;377(6612):1290-1298. doi: 
            10.1126/science.abg6621. Epub 2022 Aug 25.
  PUBMED    36007018
REFERENCE   2  (bases 1 to 13644)
  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: 
            "10.1126/science.abg6621"; journalName: "Science"; date: 
            "2022-09-16- 16"; volume: "377"; issue: "6612"; pages: "1290-1298"
COMMENT     Sequence Label: pLVX NPC1-FLAG-TiD
FEATURES             Location/Qualifiers
     source          1..13644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    673..798
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1295..1528
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1713..1757
                     /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    2020..2135
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2329..3511
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2560..3502
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3459..3479
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             7462..8418
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             8419..8442
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        8471..8970
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8506..8529)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8903..8922
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             8991..9590
                     /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(8991..9010)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9487..9507
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9604..10192
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9657..9677)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10075..10086)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10399..11032
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11161..11177)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11161..11177)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11174..11196)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11185..11201
                     /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(11209..11215,11216..11233,11234..11239))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11254..11275
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11392..11409)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11563..12151)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11643..11662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12322..13113,13114..13182))
                     /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"
     promoter        complement(13183..13287)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13335..13469
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13372..13391)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13426..13445
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             join(13637..13644,1..626)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ctaattcact cccaaagaag acaagatatc cttgatctgt ggatctacca cacacaaggc
       61 tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt
      121 ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc caataaagga
      181 gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa
      241 gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat
      301 ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac
      361 tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata
      421 taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga
      481 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
      541 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
      601 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
      661 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
      721 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
      781 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
      841 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
      901 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
      961 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     1021 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     1081 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     1141 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     1201 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     1261 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     1321 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     1381 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     1441 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     1501 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     1561 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     1621 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     1681 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     1741 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     1801 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     1861 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     1921 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     1981 gatccattcg attagtgaac ggatctcgac ggtatcgcct ttaaaagaaa aggggggatt
     2041 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2101 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     2161 gatccagttt atcgatgagt aattcataca aaaggactcg cccctgcctt ggggaatccc
     2221 agggaccgtc gttaaactcc cactaacgta gaacccagag atcgctgcgt tcccgccccc
     2281 tcacccgccc gctctcgtca tcactgaggt ggagaagagc atgcgtgagg ctccggtgcc
     2341 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     2401 aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     2461 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     2521 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc
     2581 cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacgccc
     2641 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
     2701 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcttgggcg
     2761 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
     2821 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     2881 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     2941 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     3001 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     3061 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     3121 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     3181 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     3241 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     3301 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     3361 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     3421 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     3481 aagttttttt cttccatttc aggtgtcgtg aggatcgcta gcgctaccgg actcagatct
     3541 cgagctcaag cttcgaattg gatccacgtg gaattcgcca ccatgaccgc tagagggctg
     3601 gctctggggc tgctgctgct gctgctgtgc cccgctcagg tgttcagtca gtcttgcgtg
     3661 tggtatggcg agtgcggcat cgcctatggg gacaagaggt ataactgtga atactctgga
     3721 ccccctaagc cactgcccaa agacggctac gatctggtgc aggaactgtg cccaggattc
     3781 tttttcggca acgtgagcct gtgctgtgac gtgcggcagc tgcagaccct gaaggataat
     3841 ctgcagctgc ctctgcagtt tctgtccagg tgcccatctt gtttttataa cctgctgaat
     3901 ctgttctgcg agctgacctg tagtccaagg cagtcacagt tcctgaacgt gactgccacc
     3961 gaggactacg tcgatcccgt gacaaaccag acaaagacta atgtcaaaga actgcagtac
     4021 tatgtgggcc agtccttcgc caacgctatg tataatgctt gccgagacgt ggaggcacca
     4081 agctccaatg ataaggccct ggggctgctg tgcggaaaag acgcagatgc ctgtaacgct
     4141 accaattgga tcgaatacat gtttaacaag gataatggcc aggccccttt caccattaca
     4201 ccagtgttta gcgacttccc cgtccacggg atggagccta tgaacaatgc cactaaggga
     4261 tgcgacgagt ccgtcgatga agtgaccgct ccttgctcct gtcaggattg ctctatcgtg
     4321 tgcgggccta aaccacagcc accacctcca ccagcaccat ggacaattct gggactggac
     4381 gccatgtatg tgattatgtg gatcacttac atggcctttc tgctggtctt tttcggcgca
     4441 tttttcgccg tgtggtgtta tcgaaagcgg tactttgtga gcgagtacac ccccattgac
     4501 tccaacatcg ctttcagcgt gaatgcatcc gacaaaggcg aggcatcctg ctgtgatccc
     4561 gtgagcgccg ctttcgaagg gtgcctgcgg agactgttta ctcgatgggg gtctttctgc
     4621 gtgcggaacc ccggatgcgt gatcttcttc agcctggtgt ttatcaccgc ctgttctagt
     4681 gggctggtgt tcgtccgagt gaccacaaat ccagtggatc tgtggagcgc cccatcaagc
     4741 caggctcgac tggagaagga atactttgac cagcatttcg gaccattttt cagagcagag
     4801 cagctgatca ttagggcacc cctgactgat aaacacatct accagcccta tccttctgga
     4861 gccgacgtgc ctttcggccc tccactggat atccagattc tgcatcaggt gctggacctg
     4921 cagatcgcca ttgagaacat taccgctagc tatgacaatg aaactgtgac cctgcaggat
     4981 atctgcctgg ctcctctgag cccatataac actaattgta ccattctgtc cgtcctgaac
     5041 tactttcaga atagccactc cgtgctggat cataagaaag gcgacgactt cttcgtgtac
     5101 gccgactatc acacccattt cctgtactgc gtgcgggctc ctgcaagtct gaacgacacc
     5161 tcactgctgc acgatccatg tctggggaca tttggcggac ccgtgttccc ttggctggtc
     5221 ctgggaggct atgacgatca gaactacaac aatgccaccg ccctggtcat caccttccct
     5281 gtcaacaatt actataatga taccgagaag ctgcagagag cacaggcctg ggagaaagag
     5341 ttcatcaact tcgtgaagaa ttacaaaaac ccaaatctga ccattagttt cacagccgag
     5401 cggagcatcg aggacgaact gaaccgagaa tctgacagtg acgtgttcac cgtggtcatt
     5461 agctatgcta tcatgttcct gtacatctca ctggcactgg gccatatgaa gagctgcagg
     5521 cgcctgctgg tggattcaaa agtcagcctg ggaattgccg gcatcctgat tgtgctgtcc
     5581 tctgtcgctt gtagcctggg ggtgttctcc tatattggac tgcctctgac cctgatcgtc
     5641 attgaagtga tcccatttct ggtcctggct gtcggagtgg acaatatctt cattctggtg
     5701 caggcatacc agcgcgacga gcgactgcag ggcgaaacac tggatcagca gctgggaagg
     5761 gtgctgggag aggtcgctcc atctatgttt ctgagttcat tcagtgaaac cgtggcattt
     5821 ttcctgggcg ccctgagtgt gatgccagca gtccacacat tttcactgtt cgctgggctg
     5881 gcagtgttta ttgacttcct gctgcagatc acctgctttg tgagcctgct gggcctggac
     5941 attaagaggc aggagaaaaa ccgcctggat atcttctgct gcgtgcgggg agcagaggac
     6001 ggcacatccg tgcaggcttc agaaagctgt ctgttcaggt ttttcaagaa cagctattct
     6061 ccactgctgc tgaaagactg gatgcgcccc atcgtgattg caatctttgt cggagtgctg
     6121 agtttctcaa ttgccgtgct gaacaaggtc gacatcggcc tggatcagag cctgtccatg
     6181 cccgacgatt cttacatggt ggattacttt aaatctatca gtcagtatct gcacgccggc
     6241 ccccctgtct acttcgtgct ggaggaaggc catgactaca ctagctccaa ggggcagaat
     6301 atggtgtgcg ggggaatggg atgtaacaat gattccctgg tgcagcagat cttcaacgca
     6361 gcccagctgg acaattatac aaggattggc tttgccccct ctagttggat cgacgattac
     6421 ttcgattggg tcaagcctca gtcaagctgc tgtcgcgtgg acaacatcac agatcagttt
     6481 tgcaatgcaa gcgtggtcga ccctgcctgc gtgagatgta ggccactgac tccagagggc
     6541 aaacagcgac cacagggagg cgacttcatg agattcctgc ctatgttcct gtccgacaac
     6601 ccaaatccca agtgtggcaa aggaggccac gctgcatact cctctgcagt caacatcctg
     6661 ctgggacatg ggacccgagt gggagccaca tacttcatga cataccacac tgtgctgcag
     6721 acctcagctg acttcattga tgcactgaag aaagcccgcc tgatcgctag caacgtgaca
     6781 gagactatgg ggatcaatgg atccgcctat cgagtgtttc cctactctgt cttttacgtg
     6841 ttctatgaac agtacctgac tatcattgac gataccatct tcaacctggg ggtgtccctg
     6901 ggagctatct tcctggtcac tatggtgctg ctgggctgcg agctgtggtc tgccgtgatt
     6961 atgtgcgcca ccatcgctat ggtcctggtg aatatgttcg gcgtgatgtg gctgtggggg
     7021 atctctctga acgccgtcag tctggtgaat ctggtcatgt cttgcgggat tagtgtggag
     7081 ttttgtagcc atatcactag agctttcacc gtctcaatga agggaagccg ggtggaaaga
     7141 gcagaggaag ctctggcaca catgggaagt tcagtgttta gtggcattac cctgacaaag
     7201 ttcgggggaa tcgtggtcct ggcctttgct aagagccaga tcttccagat cttttatttc
     7261 cggatgtacc tggctatggt gctgctgggc gcaacacacg ggctgatttt cctgcccgtc
     7321 ctgctgtcat atatcggccc tagcgtgaac aaggcaaaaa gctgcgccac tgaggaaaga
     7381 tacaagggga ctgagcgaga gagactgctg aactttgcgg ccgctggcgg tggtagcggt
     7441 ggcggaagcg gtggaggcag taaagacaat actgtgcctc tgaagctgat cgctctcctg
     7501 gctaatggcg agttccatag tggcgaacag ctgggagaaa ccctgggcat gtccagggcc
     7561 gctatcaaca agcacattca gactctgcgc gactggggcg tggacgtgtt caccgtgccc
     7621 ggaaagggct actctctgcc cgagcctatc ccgctgctga acgctaaaca gattctggga
     7681 cagctggacg gcgggagcgt ggcagtcctg cctgtggtcg actccaccaa tcagtacctg
     7741 ctggatcgaa tcggcgagct gaagagtggg gatgcttgca ttgcagaata tcagcaggca
     7801 gggagaggaa gcagagggag gaaatggttc tctccttttg gagctaacct gtacctgagt
     7861 atgttttggc gcctgaagcg gggaccagca gcaatcggcc tgggcccggt catcggaatt
     7921 gtcatggcag aagcgctgcg aaagctggga gcagacaagg tgcgagtcaa atggcccaat
     7981 gacctgtatc tgcaggatag aaagctggca ggcatcctgg tggagctggc cggaataaca
     8041 ggcgatgctg cacagatcgt cattggcgcc gggattaacg tggctatgag gcgcgtggag
     8101 gaaagcgtgg tcaatcaggg ctggatcaca ctgcaggaag cagggattaa cctggacagg
     8161 aatactctgg ccgctacgct gatccgagag ctgcgggcag ccctggaact gttcgagcag
     8221 gaaggcctgg ctccatatct gccacggtgg gagaagctgg ataacttcat caatagaccc
     8281 gtgaagctga tcattgggga caaagagatt ttcgggatta gccgggggat tgataaacag
     8341 ggagccctgc tgctggaaca ggacggagtt atcaaaccct ggatgggcgg agaaatcagt
     8401 ctgcggtctg ccgaaaagga ctacaaggat gacgatgata aataacccgg ctcatctaga
     8461 taattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca
     8521 gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc
     8581 accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct
     8641 cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg
     8701 gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg
     8761 gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg
     8821 ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc
     8881 cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg
     8941 ttctcctctt cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt
     9001 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     9061 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     9121 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     9181 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     9241 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     9301 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     9361 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     9421 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     9481 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     9541 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ccgcgtctgg
     9601 aacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     9661 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     9721 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     9781 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     9841 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     9901 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     9961 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
    10021 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
    10081 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
    10141 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt
    10201 aattctgcag tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
    10261 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
    10321 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
    10381 taaaagaaaa gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
    10441 atctgtggat ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc
    10501 caggggtcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag
    10561 ataaggtaga agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc
    10621 atgggatgga tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat
    10681 ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt
    10741 gctacaaggg actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg
    10801 agtggcgagc cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc
    10861 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
    10921 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
    10981 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
    11041 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
    11101 aggccttgac attgctagcg ttttaccgtc gacctctagc tagagcttgg cgtaatcatg
    11161 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
    11221 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
    11281 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
    11341 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
    11401 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
    11461 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
    11521 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
    11581 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
    11641 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
    11701 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
    11761 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
    11821 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
    11881 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
    11941 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
    12001 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
    12061 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
    12121 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
    12181 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
    12241 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
    12301 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
    12361 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
    12421 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
    12481 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
    12541 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
    12601 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
    12661 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
    12721 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
    12781 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
    12841 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
    12901 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
    12961 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
    13021 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
    13081 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
    13141 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
    13201 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
    13261 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
    13321 cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
    13381 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
    13441 tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc
    13501 ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt catttactct
    13561 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta
    13621 ctacaaactt agtagttgga aggg
//
