LOCUS       sequence-440951-       13022 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA targeting human KIAA0141/DELE1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13022)
  AUTHORS   Kenny TC, Khan A, Son Y, Yue L, Heissel S, Sharma A, Pasolli HA, Liu
            Y, Gamazon ER, Alwaseem H, Hite RK, Birsoy K
  TITLE     Integrative genetic analysis identifies FLVCR1 as a plasma-membrane 
            choline transporter in mammals.
  JOURNAL   Cell Metab. 2023 Jun 6;35(6):1057-1071.e12. doi: 
            10.1016/j.cmet.2023.04.003. Epub 2023 Apr 25.
  PUBMED    37100056
REFERENCE   2  (bases 1 to 13022)
  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"; doi: 
            "10.1016/j.cmet.2023.04.003"; journalName: "Cell Metab"; date: 
            "2023-06-6- 6"; volume: "35"; issue: "6"; pages: "1057-1071.e12"
COMMENT     Sequence Label: plentiCRISPR-v2 Puro KIAA0141/DELE1_sg1
FEATURES             Location/Qualifiers
     source          1..13022
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1062..1295
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1480..1524
                     /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"
     misc_feature    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1990..2238
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1990..2010
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2161..2180
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        2406..2617
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2636..2645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2642..6745
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2642..6745
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6746..6793
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6794..6817
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6827..6883
                     /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             6884..7480
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7377..7397
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7496..8084
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7549..7569)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7967..7978)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8156..8389
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8415..8432)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8421..8645
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8691..9119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8778..8797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8988..9009
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9128..9148)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9133..9462
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9313..9448
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9375..9394
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9510..9557
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9576..9950
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    10080..10201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10117..10136)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10171..10190
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10250..10266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10250..10266)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10263..10285)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10274..10290
                     /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)."
     promoter        complement(join(10298..10304,10305..10322,10323..10328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10343..10364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10481..10498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10652..11240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10732..10751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11411..12202,12203..12271))
                     /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     12034..12053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12272..12376)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12451..12470)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12642..13021
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taaggtaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     2041 gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg
     2101 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     2161 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
     2221 tggaaaggac gaaacaccga gcagtgttct acccgctggt ttaagagcta tgctggaaac
     2281 agcatagcaa gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     2341 gtgctttttt gaattcgcta gctaggtctt gaaaggagtg ggaattggct ccggtgcccg
     2401 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     2461 ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     2521 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     2581 cgttcttttt cgcaacgggt ttgccgccag aacacaggac cggttctaga gcgctgccac
     2641 catggacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
     2701 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     2761 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     2821 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     2881 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     2941 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     3001 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     3061 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     3121 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     3181 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     3241 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     3301 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     3361 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     3421 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     3481 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     3541 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     3601 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     3661 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     3721 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     3781 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     3841 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     3901 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     3961 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     4021 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     4081 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     4141 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     4201 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     4261 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     4321 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     4381 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     4441 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     4501 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     4561 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     4621 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     4681 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     4741 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     4801 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     4861 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     4921 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     4981 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     5041 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     5101 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca
     5161 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     5221 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     5281 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     5341 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     5401 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     5461 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     5521 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     5581 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     5641 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     5701 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     5761 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     5821 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     5881 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     5941 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     6001 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     6061 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     6121 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     6181 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     6241 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
     6301 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     6361 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     6421 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     6481 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     6541 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     6601 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     6661 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
     6721 cgacctgtct cagctgggag gcgacaagcg acctgccgcc acaaagaagg ctggacaggc
     6781 taagaagaag aaagattaca aagacgatga cgataaggga tccggcgcaa caaacttctc
     6841 tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccgaccgagt acaagcccac
     6901 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     6961 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     7021 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     7081 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     7141 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     7201 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     7261 cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     7321 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     7381 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     7441 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga acgcgttaag tcgacaatca
     7501 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     7561 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     7621 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     7681 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     7741 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     7801 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     7861 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     7921 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     7981 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     8041 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca
     8101 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     8161 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     8221 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     8281 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     8341 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta
     8401 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     8461 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     8521 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     8581 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     8641 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     8701 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     8761 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     8821 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     8881 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     8941 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     9001 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     9061 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     9121 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     9181 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     9241 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     9301 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     9361 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     9421 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     9481 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
     9541 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc
     9601 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     9661 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     9721 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     9781 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     9841 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     9901 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac
     9961 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
    10021 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
    10081 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
    10141 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
    10201 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
    10261 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
    10321 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
    10381 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
    10441 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
    10501 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
    10561 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
    10621 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
    10681 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
    10741 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
    10801 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
    10861 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
    10921 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
    10981 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
    11041 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
    11101 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
    11161 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
    11221 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
    11281 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
    11341 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
    11401 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
    11461 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
    11521 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
    11581 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
    11641 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
    11701 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
    11761 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
    11821 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
    11881 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
    11941 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
    12001 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
    12061 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    12121 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    12181 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    12241 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    12301 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
    12361 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
    12421 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
    12481 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
    12541 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
    12601 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
    12661 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
    12721 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
    12781 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
    12841 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
    12901 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
    12961 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
    13021 gg
//
