LOCUS       sequence-324916-       14128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of BE3.9max and customizable gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14128)
  AUTHORS   Hanna RE, Hegde M, Fagre CR, DeWeirdt PC, Sangree AK, Szegletes Z, 
            Griffith A, Feeley MN, Sanson KR, Baidi Y, Koblan LW, Liu DR, Neal 
            JT, Doench JG
  TITLE     Massively parallel assessment of human variants with base editor 
            screens.
  JOURNAL   Cell. 2021 Feb 18;184(4):1064-1080.e20. doi: 
            10.1016/j.cell.2021.01.012.
  PUBMED    33606977
REFERENCE   2  (bases 1 to 14128)
  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.cell.2021.01"; journalName: "Cell"; date: "2021-02-18- 
            18"; volume: "184"; issue: "4"; pages: "1064-1080.e20"
COMMENT     Sequence Label: pRDA_256
FEATURES             Location/Qualifiers
     source          1..14128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     108..128
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             171..351
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    398..523
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1016..1249
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1434..1478
                     /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    1776..1893
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1944..2192
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1944..1964
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2115..2134
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2214..2289
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2373..2584
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2634..2654
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2655..3338
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             join(3435..3458,3459..3461,3462..7535)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3435..7535
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7566..7814
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7857..7877
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7887..7943
                     /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             7944..8540
                     /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     8437..8457
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8556..9144
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8609..8629)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9027..9038)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9216..9449
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9475..9492)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9481..9705
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9751..10179
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9838..9857)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10048..10069
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10188..10208)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10193..10522
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10373..10508
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10435..10454
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10570..10617
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10636..11010
                     /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    11140..11261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11177..11196)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11231..11250
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11310..11326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11310..11326)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11323..11345)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11334..11350
                     /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(11358..11364,11365..11382,11383..11388))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11403..11424
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11541..11558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11712..12300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11792..11811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12471..13262,13263..13331))
                     /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     13094..13113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13332..13436)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13511..13530)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        13702..14081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(14083..14128,1..153)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
       61 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      121 taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct
      181 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
      241 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
      301 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
      361 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
      421 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      481 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      541 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      601 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      661 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      721 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      781 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      841 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
      901 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
      961 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1021 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1081 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1141 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1201 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1261 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1321 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     1381 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     1441 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     1501 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     1561 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     1621 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     1681 gtggagagag agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc
     1741 caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     1801 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     1861 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     1921 ccagtttggt taattaaggt accgagggcc tatttcccat gattccttca tatttgcata
     1981 tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat
     2041 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     2101 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     2161 ctttatatat cttgtggaaa ggacgaaaca ccggagacgc ttttttcgtc tcagtttgag
     2221 agctagaaat agcaagttca aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     2281 agtcggtgcg ctcacctatt agcggctaag gttttttgaa ttcgctagct aggtcttgaa
     2341 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     2401 cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt ggcgcggggt
     2461 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     2521 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     2581 acaggaccgg tgccaccatg aaacggacag ccgacggaag cgagttcgag tcaccaaaga
     2641 agaagcggaa agtctcctca gagactgggc ctgtcgccgt cgatccaacc ctgcgccgcc
     2701 ggattgaacc tcacgagttt gaagtgttct ttgacccccg ggagctgaga aaggagacat
     2761 gcctgctgta cgagatcaac tggggaggca ggcactccat ctggaggcac acctctcaga
     2821 acacaaataa gcacgtggag gtgaacttca tcgagaagtt taccacagag cggtacttct
     2881 gccccaatac cagatgtagc atcacatggt ttctgagctg gtccccttgc ggagagtgta
     2941 gcagggccat caccgagttc ctgtccagat atccacacgt gacactgttt atctacatcg
     3001 ccaggctgta tcaccacgca gacccaagga ataggcaggg cctgcgcgat ctgatcagct
     3061 ccggcgtgac catccagatc atgacagagc aggagtccgg ctactgctgg cggaacttcg
     3121 tgaattattc tcctagcaac gaggcccact ggcctaggta cccacacctg tgggtgcgcc
     3181 tgtacgtgct ggagctgtat tgcatcatcc tgggcctgcc cccttgtctg aatatcctgc
     3241 ggagaaagca gccccagctg accttcttta caatcgccct gcagtcttgt cactatcaga
     3301 ggctgccacc ccacatcctg tgggccacag gcctgaagtc tggaggatct agcggaggat
     3361 cctctggcag cgagacacca ggaacaagcg agtcagcaac accagagagc agtggcggca
     3421 gcagcggcgg cagcgacaag aagtacagca tcggcctggc catcggcacc aactctgtgg
     3481 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
     3541 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
     3601 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
     3661 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
     3721 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
     3781 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
     3841 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
     3901 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     3961 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     4021 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     4081 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     4141 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     4201 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     4261 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     4321 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     4381 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     4441 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     4501 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     4561 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     4621 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     4681 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     4741 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     4801 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     4861 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     4921 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     4981 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     5041 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     5101 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     5161 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     5221 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     5281 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     5341 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     5401 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     5461 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     5521 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     5581 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     5641 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     5701 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     5761 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     5821 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     5881 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     5941 atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     6001 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     6061 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     6121 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     6181 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     6241 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     6301 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     6361 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     6421 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     6481 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     6541 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     6601 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     6661 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     6721 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     6781 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
     6841 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     6901 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     6961 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     7021 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     7081 ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     7141 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     7201 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
     7261 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
     7321 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     7381 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
     7441 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     7501 agacacggat cgacctgtct cagctgggag gtgacagcgg cgggagcggc gggagcgggg
     7561 ggagcactaa tctgagcgac atcattgaga aggagactgg gaaacagctg gtcattcagg
     7621 agtccatcct gatgctgcct gaggaggtgg aggaagtgat cggcaacaag ccagagtctg
     7681 acatcctggt gcacaccgcc tacgacgagt ccacagatga gaatgtgatg ctgctgacct
     7741 ctgacgcccc cgagtataag ccttgggccc tggtcatcca ggattctaac ggcgagaata
     7801 agatcaagat gctgagcgga ggctccaaaa gaaccgccga cggcagcgag ttcgagccca
     7861 agaagaagag gaaagtcagc gctggcgcaa caaacttctc tctgctgaaa caagccggag
     7921 atgtcgaaga gaatcctgga ccgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     7981 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     8041 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     8101 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     8161 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     8221 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     8281 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc
     8341 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     8401 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     8461 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     8521 cccgcaagcc cggtgcctga acgcgttaag tcgacaatca acctctggat tacaaaattt
     8581 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     8641 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     8701 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     8761 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     8821 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     8881 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     8941 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     9001 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     9061 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     9121 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
     9181 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     9241 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     9301 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     9361 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     9421 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
     9481 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     9541 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     9601 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     9661 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     9721 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     9781 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     9841 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     9901 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     9961 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
    10021 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
    10081 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
    10141 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
    10201 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
    10261 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
    10321 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
    10381 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
    10441 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
    10501 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
    10561 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
    10621 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
    10681 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
    10741 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
    10801 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
    10861 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
    10921 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
    10981 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
    11041 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
    11101 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
    11161 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
    11221 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
    11281 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
    11341 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
    11401 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
    11461 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
    11521 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
    11581 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
    11641 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
    11701 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
    11761 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
    11821 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
    11881 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
    11941 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
    12001 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
    12061 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
    12121 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
    12181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
    12241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
    12301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
    12361 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
    12421 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
    12481 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
    12541 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
    12601 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
    12661 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
    12721 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
    12781 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
    12841 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
    12901 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
    12961 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
    13021 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    13081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    13141 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    13201 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    13261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    13321 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    13381 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    13441 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
    13501 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
    13561 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
    13621 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
    13681 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
    13741 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
    13801 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
    13861 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
    13921 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
    13981 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
    14041 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
    14101 acatcaatgg gcgtggatag cggtttga
//
