LOCUS       sequence-416903-        9248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct expressing FLAG Metap2 and shRNA against 
            LYCHOS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9248)
  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 9248)
  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: pLKO shLYCHOS/FLAG Metap2
FEATURES             Location/Qualifiers
     source          1..9248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     493..512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     746..763
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    880..901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(916..921,922..939,940..946)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    954..970
                     /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     959..981
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     978..994
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     978..994
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1013..1033
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1015..1033
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1061..1287
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1288..1468
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1515..1640
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2133..2366
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2551..2595
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        2893..3141
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2893..2913
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3064..3083
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    3238..3355
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3404..3914
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3833..3851
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      3923..3932
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3932..3955
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    5507..6080
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5674..5691)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5901..5920
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6100..6699
                     /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(6100..6119)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6596..6616
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6827..7060
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7132..7253
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7169..7188)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7223..7242
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7293..7428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7448..7467)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7449..7467)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7477..7494)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7477..7493)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7486..7508)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7635..8090
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7722..7741)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7932..7953
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8116..8220
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8221..8289,8290..9081)
                     /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(8439..8458)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
       61 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
      121 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
      181 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
      241 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
      301 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
      361 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
      421 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      481 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      541 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      601 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      661 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      721 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      781 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      841 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      901 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      961 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     1021 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
     1081 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
     1141 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
     1201 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
     1261 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
     1321 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     1381 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     1441 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     1501 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     1561 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     1621 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     1681 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     1741 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     1801 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     1861 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     1921 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     1981 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     2041 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     2101 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     2161 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     2221 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     2281 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     2341 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     2401 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     2461 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     2521 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     2581 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     2641 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     2701 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     2761 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     2821 tccattcgat tagtgaacgg atctcgacgg tatcgatcac gagactagcc tcgagcggcc
     2881 gcccccttca ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag
     2941 gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat
     3001 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     3061 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
     3121 ttgtggaaag gacgaaacac cggcttaacc attgcagtca atatctcgag atattgactg
     3181 caatggttaa gtttttgaat tctcgacctc gagacaaatg gcagtattca tccacaattt
     3241 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     3301 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     3361 ttattacagg gacagcagag atccactttg gccgcggctc gagggggttg gggttgcgcc
     3421 ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga
     3481 aacgcagcgg cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc
     3541 ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc
     3601 gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca
     3661 ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg
     3721 ctgtggccaa tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg
     3781 tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg
     3841 cattctgcaa gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg
     3901 acctctctcc ccagggggat ccgccaccat ggactacaag gatgacgatg acaagggtgg
     3961 tggtggtggt gcgtcgacga tggcgggtgt ggaggaggta gcggcctccg ggagccacct
     4021 gaatggcgac ctggatccag acgacaggga agaaggagct gcctctacgg ctgaggaagc
     4081 agccaagaaa aaaagacgaa agaagaagaa gagcaaaggg ccttctgcag gggaacagga
     4141 acctgataaa gaatcaggag cctcagtgga tgaagtagca agacagttgg aaagatcagc
     4201 attggaagat aaagaaagag atgaagatga tgaagatgga gatggcgatg gagatggagc
     4261 aactggaaag aagaagaaaa agaagaagaa gaagagagga ccaaaagttc aaacagaccc
     4321 tccctcagtt ccaatatgtg acctgtatcc taatggtgta tttcccaaag gacaagaatg
     4381 cgaataccca cccacacaag atgggcgaac agctgcttgg agaactacaa gtgaagaaaa
     4441 gaaagcatta gatcaggcaa gtgaagagat ttggaatgat tttcgagaag ctgcagaagc
     4501 acatcgacaa gttagaaaat acgtaatgag ctggatcaag cctgggatga caatgataga
     4561 aatctgtgaa aagttggaag actgttcacg caagttaata aaagagaatg gattaaatgc
     4621 aggcctggca tttcctactg gatgttctct caataattgt gctgcccatt atactcccaa
     4681 tgccggtgac acaacagtat tacagtatga tgacatctgt aaaatagact ttggaacaca
     4741 tataagtggt aggattattg actgtgcttt tactgtcact tttaatccca aatatgatac
     4801 gttattaaaa gctgtaaaag atgctactaa cactggaata aagtgtgctg gaattgatgt
     4861 tcgtctgtgt gatgttggtg aggccatcca agaagttatg gagtcctatg aagttgaaat
     4921 agatgggaag acatatcaag tgaaaccaat ccgtaatcta aatggacatt caattgggca
     4981 atatagaata catgctggaa aaacagtgcc gattgtgaaa ggaggggagg caacaagaat
     5041 ggaggaagga gaagtatatg caattgaaac ctttggtagt acaggaaaag gtgttgttca
     5101 tgatgatatg gaatgttcac attacatgaa aaattttgat gttggacatg tgccaataag
     5161 gcttccaaga acaaaacact tgttaaatgt catcaatgaa aactttggaa cccttgcctt
     5221 ctgccgcaga tggctggatc gcttgggaga aagtaaatac ttgatggctc tgaagaatct
     5281 gtgtgacttg ggcattgtag atccatatcc accattatgt gacattaaag gatcatatac
     5341 agcgcaattt gaacatacca tcctgttgcg tccaacatgt aaagaagttg tcagcagagg
     5401 agatgactat taatctagaa tcagataatg ggaacattat gatccagaaa cacgatggca
     5461 tcacggtggc agtgcacaaa atggcctctt gatctagacc cgggcgcccc tctccctccc
     5521 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     5581 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     5641 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     5701 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     5761 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     5821 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     5881 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     5941 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     6001 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     6061 tttgaaaaac acgatgataa agatccaccg gagcttacca tgaccgagta caagcccacg
     6121 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     6181 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     6241 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     6301 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     6361 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     6421 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     6481 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     6541 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     6601 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     6661 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag
     6721 cgcccgaccg aaaggagcgc acgaccccat gcatcggtac ctttaagacc aatgacttac
     6781 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     6841 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     6901 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     6961 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     7021 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
     7081 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
     7141 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     7201 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     7261 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     7321 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     7381 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc
     7441 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     7501 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     7561 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     7621 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     7681 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     7741 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     7801 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     7861 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     7921 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     7981 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     8041 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     8101 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     8161 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     8221 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     8281 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     8341 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     8401 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     8461 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     8521 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     8581 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     8641 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     8701 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     8761 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     8821 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     8881 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     8941 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     9001 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     9061 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     9121 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     9181 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     9241 aaaggatc
//
