LOCUS       sequence-211491-       11590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  genetic depletion of SLC19A1.
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 sgSLC19A1
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    515..536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(551..556,557..574,575..581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    589..605
                     /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     594..616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     613..629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     613..629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     648..668
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        650..668
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        696..922
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             923..1103
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1150..1275
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1768..2001
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2186..2230
                     /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        2408..2656
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2408..2428
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2579..2598
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2678..2753
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2801..2918
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2987..3198
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3218..3227
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3227..3250
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3257..3277
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3302..7402
                     /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             3302..7402
                     /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             7403..7450
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7466..7522
                     /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             7523..8122
                     /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(7523..7542)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8019..8039
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8138..8726
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8191..8211)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8609..8620)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8798..9031
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9109..9230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9146..9165)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9200..9219
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9270..9405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9425..9444)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9426..9444)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9454..9471)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9454..9470)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9463..9485)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9612..10067
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9699..9718)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9909..9930
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10093..10197
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10198..10266,10267..11058)
                     /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(10416..10435)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(11229..11590,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      661 actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta
      721 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg
      781 tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac
      841 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa
      901 gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct
      961 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     1021 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     1081 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     1141 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     1201 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     1261 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     1321 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     1381 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     1441 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     1501 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     1561 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
     1621 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     1681 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     1741 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     1801 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     1861 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     1921 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     1981 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     2041 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     2101 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     2161 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     2221 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     2281 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     2341 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     2401 acccagagag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt
     2461 tagagagata attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg
     2521 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga
     2581 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg
     2641 gaaaggacga aacaccgcga gttgaagacc caccagagtt ttagagctag aaatagcaag
     2701 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg
     2761 aattctagat cttgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     2821 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     2881 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     2941 gagatccact ttggcgccgg ctcgagtggc tccggtgccc gtcagtgggc agagcgcaca
     3001 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
     3061 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     3121 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     3181 tttgccgcca gaacacaggt gtcgtgacgc gggatccgcc accatggatt acaaagacga
     3241 tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc
     3301 cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat
     3361 caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca
     3421 cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc
     3481 cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta
     3541 tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct tccacagact
     3601 ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa
     3661 catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa
     3721 actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat
     3781 gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt
     3841 ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat
     3901 caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg
     3961 gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct
     4021 gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga
     4081 tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca
     4141 gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct
     4201 gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat
     4261 gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca
     4321 gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg
     4381 ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga
     4441 aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa
     4501 gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc
     4561 cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga
     4621 gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag
     4681 attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt
     4741 ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa
     4801 cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta
     4861 taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag
     4921 cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt
     4981 gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc
     5041 cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat
     5101 caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct
     5161 gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca
     5221 cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag
     5281 gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga
     5341 tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag
     5401 cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca
     5461 cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt
     5521 gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat
     5581 cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat
     5641 gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt
     5701 ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga
     5761 tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat
     5821 cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga
     5881 caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa
     5941 ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac
     6001 caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct
     6061 ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac
     6121 taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa
     6181 gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta
     6241 ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta
     6301 ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat
     6361 gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa
     6421 catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc
     6481 tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc
     6541 caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca
     6601 gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc
     6661 cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta
     6721 ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa
     6781 agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt
     6841 tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta
     6901 ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca
     6961 gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca
     7021 ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca
     7081 gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat
     7141 cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc
     7201 catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc
     7261 tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga
     7321 ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga
     7381 cctgtctcag ctgggaggcg acaagcgtcc tgctgctact aagaaagctg gtcaagctaa
     7441 gaaaaagaaa gctagcggca gcggcgccac caacttcagc ctgctgaagc aggccggcga
     7501 cgtggaggag aaccccggcc ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     7561 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     7621 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     7681 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     7741 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     7801 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     7861 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga
     7921 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     7981 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     8041 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     8101 gacccgcaag cccggtgcct gaacgcgtta agtcgacaat caacctctgg attacaaaat
     8161 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     8221 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     8281 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     8341 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     8401 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     8461 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     8521 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     8581 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     8641 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     8701 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
     8761 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     8821 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     8881 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     8941 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     9001 cccttttagt cagtgtggaa aatctctagc agtacgtata gtagttcatg tcatcttatt
     9061 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     9121 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     9181 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     9241 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     9301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     9361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa
     9421 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     9481 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     9541 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     9601 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     9661 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     9721 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     9781 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     9841 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     9901 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     9961 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
    10021 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
    10081 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
    10141 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
    10201 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
    10261 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
    10321 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
    10381 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
    10441 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
    10501 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
    10561 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
    10621 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
    10681 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
    10741 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
    10801 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
    10861 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
    10921 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
    10981 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
    11041 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
    11101 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
    11161 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
    11221 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
    11281 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
    11341 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
    11401 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
    11461 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
    11521 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
    11581 cgaacgacct
//
