LOCUS       Exported               13030 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Lentivirus for constitutive expression of xCas9(3.7)-BE4Gam in 
            mammalian cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    pLenti-xBE4Gam-P2A-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13030)
  AUTHORS   Zafra MP, Schatoff EM, Katti A, Foronda M, Breinig M, Schweitzer AY,
            Simon A, Han T, Goswami S, Montgomery E, Thibado J, Kastenhuber ER, 
            Sanchez-Rivera FJ, Shi J, Vakoc CR, Lowe SW, Tschaharganeh DF, Dow 
            LE
  TITLE     Optimized base editors enable efficient editing in cells, organoids 
            and mice.
  JOURNAL   Nat Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194.
  PUBMED    29969439
REFERENCE   2  (bases 1 to 13030)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13030
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..212
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      232..241
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             241..264
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             271..291
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             883..1566
                     /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             1615..5715
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 mutant with broad PAM
                     compatibility (Hu et al., 2018)"
                     /label=xCas9-3.7
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5728..5976
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5989..6009
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6010..6258
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6271..6291
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6307..6363
                     /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             6364..6963
                     /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(6364..6383)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6860..6880
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6979..7567
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7032..7052)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7450..7461)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7639..7872
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7950..8071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7987..8006)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8041..8060
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8111..8246
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8173..8192
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(8266..8285)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8267..8285)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8295..8312)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8295..8311)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8304..8326)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8453..8908
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8540..8559)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8750..8771
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8934..9038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9039..9899
                     /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(9257..9276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10070..10658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10559..10578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10812..10829
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10946..10967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        10982..11012
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11020..11036
                     /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     11025..11047
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11044..11060
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11044..11060
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11079..11099
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11081..11099
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        11127..11353
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             11354..11534
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11581..11706
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12199..12432
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12617..12661
                     /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    12845..12962
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
       61 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
      121 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
      181 tttttcgcaa cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc cgccaccatg
      241 gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac
      301 ggagtcccag cagccgcaaa acctgcaaag agaattaaat ccgcagcagc agcctacgtg
      361 cctcaaaacc gggatgccgt tatcacagat ataaaaagaa tcggtgattt gcagcgcgaa
      421 gcaagccgct tggagaccga aatgaatgat gccatcgcag agatcactga gaaatttgct
      481 gcccgcatag caccaatcaa gactgacatc gagacactca gtaagggcgt gcaaggctgg
      541 tgcgaggcta atcgggacga gttgaccaac ggggggaagg tgaaaaccgc caatcttgtg
      601 actggcgatg tctcctggcg agtgagacca ccaagcgtaa gcatccgagg catggacgct
      661 gtgatggaaa cattggaaag gctcggcctg caaaggttta tcagaacaaa gcaggaaata
      721 aataaggaag ccatcctcct tgagccaaaa gccgttgctg gggtagccgg aattactgtt
      781 aagtctggta tcgaggattt cagtatcata cccttcgagc aggaagccgg cattagcgga
      841 agtgaaacac ccggtacctc agagagcgca actcctgaga gtagctcaga gactggccca
      901 gtggctgtgg accccacatt gagacggcgg atcgagcccc atgagtttga ggtattcttc
      961 gatccgagag agctccgcaa ggagacctgc ctgctttacg aaattaattg ggggggccgg
     1021 cactccattt ggcgacatac atcacagaac actaacaagc acgtcgaagt caacttcatc
     1081 gagaagttca cgacagaaag atatttctgt ccgaacacaa ggtgcagcat tacctggttt
     1141 ctcagctgga gcccatgcgg cgaatgtagt agggccatca ctgaattcct gtcaaggtat
     1201 ccccacgtca ctctgtttat ttacatcgca aggctgtacc accacgctga cccccgcaat
     1261 cgacaaggcc tgcgggattt gatctcttca ggtgtgacta tccaaattat gactgagcag
     1321 gagtcaggat actgctggag aaactttgtg aattatagcc cgagtaatga agcccactgg
     1381 cctaggtatc cccatctgtg ggtacgactg tacgttcttg aactgtactg catcatactg
     1441 ggcctgcctc cttgtctcaa cattctgaga aggaagcagc cacagctgac attctttacc
     1501 atcgctcttc agtcttgtca ttaccagcga ctgcccccac acattctctg ggccaccggg
     1561 ttgaaaagcg gcagcgagac tcccgggacc tcagagtccg ccacacccga aagtgacaag
     1621 aagtacagca tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac
     1681 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     1741 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
     1801 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     1861 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     1921 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     1981 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     2041 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     2101 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     2161 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     2221 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     2281 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc
     2341 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggataccaaa
     2401 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     2461 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     2521 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     2581 ctgtacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     2641 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     2701 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     2761 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     2821 accttcgaca acggcatcat cccccaccag atccacctgg gagagctgca cgccattctg
     2881 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     2941 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     3001 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagaa ggtggtggac
     3061 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     3121 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     3181 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgac
     3241 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     3301 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     3361 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     3421 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     3481 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     3541 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     3601 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     3661 aagtccgacg gcttcgccaa cagaaacttc atccagctga tccacgacga cagcctgacc
     3721 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     3781 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     3841 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     3901 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     3961 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     4021 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     4081 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
     4141 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
     4201 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     4261 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
     4321 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     4381 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     4441 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     4501 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     4561 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     4621 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     4681 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     4741 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     4801 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
     4861 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     4921 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
     4981 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     5041 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     5101 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     5161 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     5221 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgtgct gcagaaggga
     5281 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     5341 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
     5401 cactacctgg acgagatcat cgagcagatt agcgagttct ccaagagagt gatcctggcc
     5461 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
     5521 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     5581 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     5641 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     5701 cagctgggag gcgattcagg cggatctact aatctgtcag atattattga aaaggagacc
     5761 ggtaagcaac tggttatcca ggaatccatc ctcatgctcc cagaggaggt ggaagaagtc
     5821 attgggaaca agccggaaag cgatatactc gtgcacaccg cctacgacga gagcaccgac
     5881 gagaatgtca tgcttctgac tagcgacgcc cctgaataca agccttgggc tctggtcata
     5941 caggatagca acggtgagaa caagattaag atgctctctg gtggttctcc caagaagaag
     6001 aggaaagtca caaatctctc tgacatcata gagaaggaga cagggaaaca actcgtaata
     6061 caagagtcca ttcttatgct ccctgaggag gtggaagaag ttatcggcaa caaaccagag
     6121 agtgacattc tggtccatac cgcctacgat gaaagcacag acgagaacgt tatgttgctc
     6181 acttctgacg ctccagaata caaaccttgg gcactcgtca ttcaggacag caacggcgag
     6241 aacaagatca aaatgcttag cgggggcagc cccaaaaaaa agaggaaggt cgctagcggc
     6301 agcggcgcca ccaacttcag cctgctgaag caggccggcg acgtggagga gaaccccggc
     6361 cccatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     6421 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     6481 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     6541 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     6601 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     6661 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     6721 gagcccgcgt ggttcctggc caccgtcgga gtctcgcccg accaccaggg caagggtctg
     6781 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     6841 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     6901 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     6961 tgaacgcgtt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     7021 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     7081 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     7141 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     7201 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     7261 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     7321 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     7381 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     7441 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     7501 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     7561 tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     7621 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     7681 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     7741 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     7801 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     7861 aaatctctag cagtacgtat agtagttcat gtcatcttat tattcagtat ttataacttg
     7921 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     7981 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     8041 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     8101 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     8161 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     8221 gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg
     8281 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     8341 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     8401 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     8461 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     8521 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     8581 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     8641 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     8701 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     8761 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     8821 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     8881 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     8941 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     9001 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     9061 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     9121 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     9181 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     9241 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     9301 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     9361 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     9421 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     9481 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     9541 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     9601 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     9661 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     9721 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     9781 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     9841 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     9901 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     9961 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
    10021 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
    10081 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
    10141 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
    10201 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
    10261 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
    10321 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
    10381 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
    10441 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
    10501 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
    10561 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
    10621 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
    10681 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
    10741 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
    10801 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
    10861 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
    10921 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
    10981 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
    11041 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
    11101 aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt agtcttgcaa
    11161 catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg
    11221 attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac
    11281 atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc
    11341 tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
    11401 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
    11461 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
    11521 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
    11581 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
    11641 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
    11701 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
    11761 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
    11821 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
    11881 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
    11941 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
    12001 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
    12061 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
    12121 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
    12181 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
    12241 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
    12301 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
    12361 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
    12421 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
    12481 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
    12541 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
    12601 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
    12661 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
    12721 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
    12781 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccagacc
    12841 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
    12901 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
    12961 ttcgggttta ttacagggac agcagagatc cactttggcg ccggctcgag tggctccggt
    13021 gcccgtcagt
//
