LOCUS       sequence-203997-       11590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  genetic depletion of AMDHD1.
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 sgAMDHD1
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    93..114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(129..134,135..152,153..159)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    167..183
                     /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     172..194
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     191..207
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     191..207
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     226..246
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        228..246
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        274..500
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             501..681
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    728..853
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1346..1579
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1764..1808
                     /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        1986..2234
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1986..2006
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2157..2176
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2256..2331
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2379..2496
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2565..2776
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2796..2805
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2805..2828
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2835..2855
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2880..6980
                     /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             2880..6980
                     /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             6981..7028
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7044..7100
                     /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             7101..7700
                     /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(7101..7120)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7597..7617
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7716..8304
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7769..7789)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8187..8198)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8376..8609
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8687..8808
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8724..8743)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8778..8797
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8848..8983
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9003..9022)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9004..9022)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9032..9049)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9032..9048)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9041..9063)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9190..9645
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9277..9296)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9487..9508
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9671..9775
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9776..9844,9845..10636)
                     /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(9994..10013)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10807..11395
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11296..11315
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11549..11566
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
       61 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
      121 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      181 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
      241 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
      301 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
      361 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
      421 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
      481 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct
      541 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
      601 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
      661 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
      721 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
      781 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
      841 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
      901 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
      961 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     1021 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     1081 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     1141 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     1201 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     1261 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     1321 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     1381 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     1441 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     1501 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     1561 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     1621 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     1681 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     1741 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     1801 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     1861 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     1921 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     1981 ccagagaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     2041 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     2101 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     2161 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     2221 aaggacgaaa caccgatgga aattcaccag gccgggtttt agagctagaa atagcaagtt
     2281 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa
     2341 ttctagatct tgagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
     2401 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2461 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     2521 gatccacttt ggcgccggct cgagtggctc cggtgcccgt cagtgggcag agcgcacatc
     2581 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     2641 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     2701 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     2761 tgccgccaga acacaggtgt cgtgacgcgg gatccgccac catggattac aaagacgatg
     2821 acgataagat ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg
     2881 acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca
     2941 ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca
     3001 gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca
     3061 cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc
     3121 tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg
     3181 aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca
     3241 tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac
     3301 tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga
     3361 tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg
     3421 acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca
     3481 acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc
     3541 tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga
     3601 ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg
     3661 ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga
     3721 tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc
     3781 tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga
     3841 tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc
     3901 agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct
     3961 acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa
     4021 agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc
     4081 agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca
     4141 ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga
     4201 agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat
     4261 tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg
     4321 tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc
     4381 tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata
     4441 acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg
     4501 gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga
     4561 agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg
     4621 gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca
     4681 aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga
     4741 ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc
     4801 tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc
     4861 tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt
     4921 tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc
     4981 tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg
     5041 agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga
     5101 aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg
     5161 aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga
     5221 agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg
     5281 aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata
     5341 tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg
     5401 tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca
     5461 agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
     5521 actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca
     5581 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
     5641 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
     5701 agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
     5761 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
     5821 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
     5881 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
     5941 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
     6001 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
     6061 tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca
     6121 ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
     6181 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca
     6241 gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt
     6301 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
     6361 agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
     6421 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
     6481 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga
     6541 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
     6601 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
     6661 acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
     6721 tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca
     6781 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
     6841 ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg
     6901 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
     6961 tgtctcagct gggaggcgac aagcgtcctg ctgctactaa gaaagctggt caagctaaga
     7021 aaaagaaagc tagcggcagc ggcgccacca acttcagcct gctgaagcag gccggcgacg
     7081 tggaggagaa ccccggcccc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     7141 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     7201 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     7261 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     7321 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     7381 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     7441 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc
     7501 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     7561 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     7621 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     7681 cccgcaagcc cggtgcctga acgcgttaag tcgacaatca acctctggat tacaaaattt
     7741 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     7801 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     7861 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     7921 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     7981 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     8041 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     8101 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     8161 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     8221 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     8281 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
     8341 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     8401 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     8461 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     8521 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     8581 cttttagtca gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat
     8641 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     8701 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     8761 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     8821 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     8881 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     8941 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     9001 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     9061 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     9121 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     9181 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     9241 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     9301 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     9361 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     9421 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     9481 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     9541 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     9601 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     9661 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     9721 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     9781 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     9841 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     9901 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     9961 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
    10021 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
    10081 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
    10141 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
    10201 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
    10261 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
    10321 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
    10381 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
    10441 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
    10501 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
    10561 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
    10621 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
    10681 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
    10741 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
    10801 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
    10861 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
    10921 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
    10981 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
    11041 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
    11101 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
    11161 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
    11221 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
    11281 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
    11341 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
    11401 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
    11461 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
    11521 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
    11581 cccaatacgc
//
