LOCUS       sequence-203996-       11590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  genetic depletion of HAL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11590)
  AUTHORS   Kanarek N, Keys HR, Cantor JR, Lewis CA, Chan SH, Kunchok T, 
            Abu-Remaileh M, Freinkman E, Schweitzer LD, Sabatini DM
  TITLE     Histidine catabolism is a major determinant of methotrexate 
            sensitivity.
  JOURNAL   Nature. 2018 Jul 11. pii: 10.1038/s41586-018-0316-7. doi: 
            10.1038/s41586-018-0316-7.
  PUBMED    29995852
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: "Nature. 
            2018 Jul 11. pii: 10.1038/s41586-018-0316-7. doi: 
            10.1038/s41586-018-0316-7."
COMMENT     Sequence Label: pLentiCRISPR sgHAL
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..35
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    152..173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(188..193,194..211,212..218)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    226..242
                     /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     231..253
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     250..266
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     250..266
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     285..305
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        287..305
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        333..559
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             560..740
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    787..912
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1405..1638
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1823..1867
                     /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        2045..2293
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2045..2065
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2216..2235
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2315..2390
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2438..2555
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2624..2835
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2855..2864
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2864..2887
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2894..2914
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2939..7039
                     /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             2939..7039
                     /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             7040..7087
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7103..7159
                     /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             7160..7759
                     /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(7160..7179)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7656..7676
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7775..8363
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7828..7848)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8246..8257)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8435..8668
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8746..8867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8783..8802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8837..8856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8907..9042
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9062..9081)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9063..9081)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9091..9108)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9091..9107)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9100..9122)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9249..9704
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9336..9355)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9546..9567
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9730..9834
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9835..9903,9904..10695)
                     /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(10053..10072)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10866..11454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11355..11374
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
       61 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
      121 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
      181 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      241 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
      301 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
      361 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
      421 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
      481 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
      541 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
      601 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
      661 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
      721 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
      781 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
      841 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
      901 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
      961 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     1021 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     1081 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     1141 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     1201 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     1261 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     1321 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     1381 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     1441 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     1501 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     1561 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     1621 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     1681 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     1741 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     1801 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     1861 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     1921 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     1981 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     2041 cagagagggc ctatttccca tgattccttc atatttgcat atacgataca aggctgttag
     2101 agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
     2161 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
     2221 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
     2281 aggacgaaac accggcaggc agctcactga gggagtttta gagctagaaa tagcaagtta
     2341 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgaat
     2401 tctagatctt gagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     2461 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     2521 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     2581 atccactttg gcgccggctc gagtggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     2641 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     2701 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     2761 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     2821 gccgccagaa cacaggtgtc gtgacgcggg atccgccacc atggattaca aagacgatga
     2881 cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga
     2941 caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac
     3001 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag
     3061 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac
     3121 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct
     3181 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga
     3241 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat
     3301 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact
     3361 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat
     3421 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga
     3481 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa
     3541 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct
     3601 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat
     3661 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc
     3721 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat
     3781 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct
     3841 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat
     3901 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca
     3961 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta
     4021 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa
     4081 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca
     4141 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat
     4201 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa
     4261 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt
     4321 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt
     4381 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct
     4441 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa
     4501 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg
     4561 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa
     4621 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg
     4681 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa
     4741 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac
     4801 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct
     4861 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct
     4921 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt
     4981 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct
     5041 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga
     5101 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa
     5161 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga
     5221 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa
     5281 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga
     5341 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat
     5401 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt
     5461 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa
     5521 gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta
     5581 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa
     5641 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt
     5701 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa
     5761 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct
     5821 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca
     5881 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc
     5941 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat
     6001 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat
     6061 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct
     6121 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac
     6181 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac
     6241 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag
     6301 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc
     6361 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga
     6421 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct
     6481 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc
     6541 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa
     6601 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta
     6661 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca
     6721 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct
     6781 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat
     6841 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc
     6901 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt
     6961 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct
     7021 gtctcagctg ggaggcgaca agcgtcctgc tgctactaag aaagctggtc aagctaagaa
     7081 aaagaaagct agcggcagcg gcgccaccaa cttcagcctg ctgaagcagg ccggcgacgt
     7141 ggaggagaac cccggcccca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     7201 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     7261 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     7321 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     7381 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     7441 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     7501 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca
     7561 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     7621 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     7681 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     7741 ccgcaagccc ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
     7801 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     7861 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     7921 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     7981 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     8041 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     8101 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     8161 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     8221 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     8281 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     8341 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
     8401 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     8461 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     8521 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     8581 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     8641 ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt
     8701 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     8761 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     8821 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     8881 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     8941 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     9001 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
     9061 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     9121 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     9181 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     9241 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     9301 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     9361 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     9421 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     9481 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     9541 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     9601 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     9661 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     9721 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     9781 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     9841 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     9901 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     9961 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
    10021 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
    10081 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
    10141 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
    10201 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
    10261 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
    10321 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
    10381 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
    10441 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
    10501 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
    10561 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
    10621 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
    10681 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
    10741 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
    10801 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
    10861 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
    10921 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
    10981 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
    11041 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
    11101 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
    11161 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
    11221 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
    11281 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
    11341 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
    11401 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
    11461 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
    11521 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
    11581 gctcgccgca
//
