LOCUS       sequence-321913-       11590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector expressing Cas9 and an sgRNA targeting GPT2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11590)
  AUTHORS   Rossiter NJ, Huggler KS, Adelmann CH, Keys HR, Soens RW, Sabatini 
            DM, Cantor JR
  TITLE     CRISPR screens in physiologic medium reveal conditionally essential 
            genes in human cells.
  JOURNAL   Cell Metab. 2021 Feb 23. pii: S1550-4131(21)00061-9. doi: 
            10.1016/j.cmet.2021.02.005.
  PUBMED    33651980
REFERENCE   2  (bases 1 to 11590)
  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"; journalName: "Cell Metab.
            2021 Feb 23. pii: S1550-4131(21)00061-9. doi: 
            10.1016/j.cmet.2021.02.005."
COMMENT     Sequence Label: pLentiCRISPR-v1-sgGPT2_5
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     36..56
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        38..56
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        84..310
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             311..491
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    538..663
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1156..1389
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1574..1618
                     /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        1796..2044
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1796..1816
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1967..1986
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2066..2141
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2189..2306
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2375..2586
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2606..2615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2615..2638
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2645..2665
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2690..6790
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2690..6790
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6791..6838
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6854..6910
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6911..7510
                     /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(6911..6930)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7407..7427
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7526..8114
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7579..7599)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7997..8008)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8186..8419
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8497..8618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8534..8553)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8588..8607
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8658..8793
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8813..8832)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8814..8832)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8842..8859)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8842..8858)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8851..8873)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9000..9455
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9087..9106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9297..9318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9481..9585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9586..9654,9655..10446)
                     /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(9804..9823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10617..11205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11106..11125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11359..11376
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11493..11514
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11529..11534,11535..11552,11553..11559)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11567..11583
                     /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     join(11572..11590,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
       61 aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat
      121 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt
      181 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg
      241 gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg
      301 atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
      361 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
      421 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
      481 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
      541 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
      601 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
      661 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
      721 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
      781 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
      841 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
      901 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
      961 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
     1021 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     1081 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     1141 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     1201 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     1261 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     1321 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     1381 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     1441 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     1501 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     1561 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     1621 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     1681 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     1741 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccagagaggg
     1801 cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta gagagataat
     1861 tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta
     1921 ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct
     1981 taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa
     2041 caccgtgaat gtgaatgagc tccgggtttt agagctagaa atagcaagtt aaaataaggc
     2101 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa ttctagatct
     2161 tgagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca
     2221 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
     2281 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccacttt
     2341 ggcgccggct cgagtggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     2401 cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg
     2461 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     2521 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     2581 acacaggtgt cgtgacgcgg gatccgccac catggattac aaagacgatg acgataagat
     2641 ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg acaagaagta
     2701 cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     2761 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     2821 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
     2881 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     2941 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     3001 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     3061 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     3121 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     3181 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     3241 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     3301 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     3361 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag
     3421 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     3481 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     3541 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     3601 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     3661 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     3721 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     3781 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     3841 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     3901 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     3961 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     4021 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     4081 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     4141 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     4201 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     4261 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     4321 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     4381 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     4441 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     4501 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     4561 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     4621 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     4681 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     4741 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     4801 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     4861 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     4921 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     4981 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     5041 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     5101 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     5161 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag
     5221 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg
     5281 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
     5341 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
     5401 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     5461 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
     5521 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     5581 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
     5641 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
     5701 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
     5761 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
     5821 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
     5881 aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa
     5941 agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
     6001 cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga
     6061 ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt
     6121 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
     6181 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
     6241 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
     6301 gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga
     6361 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
     6421 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta
     6481 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
     6541 taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca
     6601 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa
     6661 gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc
     6721 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
     6781 gggaggcgac aagcgtcctg ctgctactaa gaaagctggt caagctaaga aaaagaaagc
     6841 tagcggcagc ggcgccacca acttcagcct gctgaagcag gccggcgacg tggaggagaa
     6901 ccccggcccc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     6961 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     7021 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     7081 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     7141 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     7201 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     7261 gcccaaggag cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc accagggcaa
     7321 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     7381 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     7441 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     7501 cggtgcctga acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     7561 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     7621 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     7681 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     7741 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     7801 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     7861 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     7921 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     7981 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     8041 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     8101 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
     8161 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     8221 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     8281 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     8341 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     8401 gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta
     8461 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     8521 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     8581 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     8641 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     8701 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     8761 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     8821 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     8881 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     8941 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     9001 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     9061 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     9121 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     9181 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     9241 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     9301 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     9361 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     9421 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     9481 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     9541 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     9601 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     9661 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     9721 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     9781 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     9841 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     9901 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     9961 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
    10021 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
    10081 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
    10141 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
    10201 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
    10261 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
    10321 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
    10381 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
    10441 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
    10501 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
    10561 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
    10621 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
    10681 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
    10741 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
    10801 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
    10861 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
    10921 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
    10981 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
    11041 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
    11101 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
    11161 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
    11221 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
    11281 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
    11341 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
    11401 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
    11461 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
    11521 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
    11581 caatttcaca
//
