LOCUS       sequence-197004-       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_2
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..228
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             229..409
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    456..581
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1074..1307
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1492..1536
                     /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        1714..1962
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1714..1734
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1885..1904
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1984..2059
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2107..2224
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2293..2504
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2524..2533
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2533..2556
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2563..2583
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2608..6708
                     /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             2608..6708
                     /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             6709..6756
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6772..6828
                     /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             6829..7428
                     /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(6829..6848)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7325..7345
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7444..8032
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7497..7517)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7915..7926)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8104..8337
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8415..8536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8452..8471)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8506..8525
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8576..8711
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8731..8750)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8732..8750)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8760..8777)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8760..8776)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8769..8791)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8918..9373
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9005..9024)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9215..9236
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9399..9503
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9504..9572,9573..10364)
                     /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(9722..9741)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10535..11123
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11024..11043
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11277..11294
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11411..11432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11447..11452,11453..11470,11471..11477)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11485..11501
                     /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     11490..11512
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11509..11525
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11509..11525
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11544..11564
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11546..11564
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat
       61 gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat
      121 cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg
      181 ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg
      241 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      301 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
      361 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
      421 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
      481 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
      541 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
      601 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
      661 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
      721 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
      781 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
      841 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
      901 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
      961 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     1021 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     1081 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     1141 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     1201 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     1261 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     1321 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     1381 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     1441 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     1501 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     1561 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     1621 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     1681 cagacccacc tcccaacccc gaggggaccc agagagggcc tatttcccat gattccttca
     1741 tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac
     1801 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca
     1861 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc
     1921 gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgggagcca tcacacagac
     1981 cccgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     2041 agtggcaccg agtcggtgct tttttgaatt ctagatcttg agacaaatgg cagtattcat
     2101 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     2161 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     2221 ttttcgggtt tattacaggg acagcagaga tccactttgg cgccggctcg agtggctccg
     2281 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
     2341 tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
     2401 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
     2461 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtgtcg tgacgcggga
     2521 tccgccacca tggattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag
     2581 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc
     2641 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     2701 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     2761 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     2821 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     2881 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     2941 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     3001 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     3061 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     3121 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     3181 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     3241 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     3301 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     3361 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     3421 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     3481 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     3541 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     3601 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     3661 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     3721 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     3781 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     3841 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     3901 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     3961 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     4021 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     4081 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     4141 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     4201 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     4261 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     4321 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     4381 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     4441 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     4501 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     4561 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     4621 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     4681 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     4741 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     4801 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     4861 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     4921 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     4981 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     5041 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     5101 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc
     5161 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     5221 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     5281 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     5341 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     5401 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     5461 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     5521 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     5581 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     5641 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     5701 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     5761 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     5821 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     5881 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     5941 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     6001 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     6061 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     6121 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     6181 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     6241 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     6301 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     6361 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     6421 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     6481 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     6541 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     6601 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     6661 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa gcgtcctgct
     6721 gctactaaga aagctggtca agctaagaaa aagaaagcta gcggcagcgg cgccaccaac
     6781 ttcagcctgc tgaagcaggc cggcgacgtg gaggagaacc ccggccccat gaccgagtac
     6841 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     6901 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     6961 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     7021 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     7081 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     7141 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     7201 ctggccaccg tcggagtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     7261 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     7321 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     7381 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc
     7441 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     7501 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     7561 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     7621 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     7681 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     7741 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     7801 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     7861 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     7921 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     7981 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt
     8041 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     8101 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     8161 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     8221 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     8281 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     8341 cgtatagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     8401 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     8461 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     8521 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     8581 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     8641 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     8701 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     8761 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     8821 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     8881 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     8941 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     9001 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     9061 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     9121 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     9181 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     9241 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     9301 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     9361 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     9421 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     9481 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     9541 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     9601 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     9661 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     9721 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     9781 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     9841 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     9901 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     9961 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
    10021 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
    10081 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
    10141 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
    10201 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
    10261 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
    10321 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
    10381 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
    10441 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
    10501 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
    10561 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
    10621 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
    10681 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
    10741 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
    10801 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
    10861 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
    10921 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
    10981 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
    11041 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
    11101 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
    11161 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
    11221 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
    11281 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
    11341 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
    11401 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
    11461 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
    11521 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
    11581 tgcaagctta
//
