LOCUS       sequence-197005-       11590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  genetic depletion of FTCD.
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 sgFTCD_1
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    445..678
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             863..907
                     /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        1085..1333
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1085..1105
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1256..1275
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1355..1430
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    1478..1595
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1664..1875
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1895..1904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1904..1927
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1934..1954
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1979..6079
                     /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             1979..6079
                     /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             6080..6127
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6143..6199
                     /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             6200..6799
                     /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(6200..6219)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6696..6716
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6815..7403
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6868..6888)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7286..7297)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7475..7708
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7786..7907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7823..7842)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7877..7896
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7947..8082
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8102..8121)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8103..8121)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8131..8148)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8131..8147)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8140..8162)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8289..8744
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8376..8395)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8586..8607
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8770..8874
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8875..8943,8944..9735)
                     /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(9093..9112)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9906..10494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10395..10414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10648..10665
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10782..10803
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10818..10823,10824..10841,10842..10848)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10856..10872
                     /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     10861..10883
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10880..10896
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10880..10896
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10915..10935
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10917..10935
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10963..11189
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             11190..11370
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11417..11542
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
       61 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
      121 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
      181 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
      241 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
      301 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
      361 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
      421 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
      481 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
      541 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
      601 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
      661 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
      721 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
      781 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
      841 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
      901 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
      961 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     1021 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     1081 cagagagggc ctatttccca tgattccttc atatttgcat atacgataca aggctgttag
     1141 agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
     1201 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
     1261 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
     1321 aggacgaaac accggcaggt tgagcgcgat gcgggtttta gagctagaaa tagcaagtta
     1381 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgaat
     1441 tctagatctt gagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     1501 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     1561 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     1621 atccactttg gcgccggctc gagtggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     1681 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     1741 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     1801 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     1861 gccgccagaa cacaggtgtc gtgacgcggg atccgccacc atggattaca aagacgatga
     1921 cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga
     1981 caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac
     2041 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag
     2101 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac
     2161 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct
     2221 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga
     2281 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat
     2341 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact
     2401 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat
     2461 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga
     2521 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa
     2581 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct
     2641 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat
     2701 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc
     2761 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat
     2821 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct
     2881 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat
     2941 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca
     3001 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta
     3061 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa
     3121 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca
     3181 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat
     3241 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa
     3301 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt
     3361 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt
     3421 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct
     3481 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa
     3541 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg
     3601 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa
     3661 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg
     3721 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa
     3781 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac
     3841 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct
     3901 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct
     3961 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt
     4021 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct
     4081 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga
     4141 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa
     4201 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga
     4261 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa
     4321 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga
     4381 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat
     4441 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt
     4501 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa
     4561 gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta
     4621 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa
     4681 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt
     4741 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa
     4801 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct
     4861 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca
     4921 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc
     4981 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat
     5041 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat
     5101 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct
     5161 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac
     5221 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac
     5281 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag
     5341 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc
     5401 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga
     5461 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct
     5521 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc
     5581 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa
     5641 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta
     5701 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca
     5761 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct
     5821 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat
     5881 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc
     5941 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt
     6001 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct
     6061 gtctcagctg ggaggcgaca agcgtcctgc tgctactaag aaagctggtc aagctaagaa
     6121 aaagaaagct agcggcagcg gcgccaccaa cttcagcctg ctgaagcagg ccggcgacgt
     6181 ggaggagaac cccggcccca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     6241 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     6301 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     6361 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     6421 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     6481 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     6541 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca
     6601 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     6661 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     6721 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     6781 ccgcaagccc ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
     6841 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     6901 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     6961 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     7021 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     7081 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     7141 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     7201 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     7261 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     7321 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     7381 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
     7441 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     7501 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     7561 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     7621 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     7681 ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt
     7741 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     7801 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     7861 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     7921 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     7981 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     8041 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
     8101 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     8161 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     8221 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     8281 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     8341 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     8401 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     8461 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     8521 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     8581 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     8641 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     8701 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     8761 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     8821 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     8881 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     8941 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     9001 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     9061 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     9121 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     9181 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     9241 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     9301 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     9361 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     9421 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     9481 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     9541 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     9601 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     9661 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     9721 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     9781 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     9841 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     9901 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     9961 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
    10021 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
    10081 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
    10141 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
    10201 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
    10261 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
    10321 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
    10381 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
    10441 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
    10501 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
    10561 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
    10621 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
    10681 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
    10741 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
    10801 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
    10861 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
    10921 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
    10981 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
    11041 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
    11101 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
    11161 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
    11221 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
    11281 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
    11341 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
    11401 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
    11461 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
    11521 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
    11581 aattcggtta
//
