LOCUS       sequence-233054-       13479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stably express BE3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13479)
  AUTHORS   Li Q, Li Y, Yang S, Huang S, Yan M, Ding Y, Tang W, Lou X, Yin Q, 
            Sun Z, Lu L, Shi H, Wang H, Chen Y, Li J
  TITLE     CRISPR-Cas9-mediated base-editing screening in mice identifies DND1 
            amino acids that are critical for primordial germ cell development.
  JOURNAL   Nat Cell Biol. 2018 Oct 1. pii: 10.1038/s41556-018-0202-4. doi: 
            10.1038/s41556-018-0202-4.
  PUBMED    30275529
REFERENCE   2  (bases 1 to 13479)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Cell 
            Biol. 2018 Oct 1. pii: 10.1038/s41556-018-0202-4. doi: 
            10.1038/s41556-018-0202-4."
COMMENT     Sequence Label: PB-BE3-NLS
FEATURES             Location/Qualifiers
     source          1..13479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   892..954
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        2038..2417
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2420..2695
                     /label=chicken beta-actin promoter
     intron          2696..3708
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3632..3654
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3716..3735
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3768..4442
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             3768..4442
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(3898..3916)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4166..4185
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4360..4383
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             4446..4499
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4506..5192
                     /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="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             join(5241..8639,8640..8642,8643..9239,9240,9241..9243,
                     9244..9245,9246..9248,9249..9341)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /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             join(5241..5264,5265..5267,5268..9341)
                     /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             9354..9602
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9615..9635
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9644..9661
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(9788..9807)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    9853..9908
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9907..9926)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(10269..10285)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    10293..10309
                     /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(10317..10323,10324..10341,10342..10347))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10362..10383
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   11084..11118
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(11291..11311)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11291..11309)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11330..11346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    11354..11370
                     /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(11378..11384,11385..11402,11403..11408))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11423..11444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11561..11578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11732..12320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11812..11831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12491..13282,13283..13351))
                     /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     13114..13133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13352..13456)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggccgc ccggtttatc gttaatatgg atcaatttga acagttgatt aacgtgtctc
      721 tgctcaagtc tttgatcaaa acgcaaatcg acgaaaatgt gtcggacaat atcaagtcga
      781 tgagcgaaaa actaaaaagg ctagaatacg acaatctcac agacagcgtt gagatatacg
      841 gtattcacga cagcaggctg aataataaaa aaattagaaa ctattattta accctagaaa
      901 gataatcata ttgtgacgta cgttaaagat aatcatgcgt aaaattgacg catgtgtttt
      961 atcggtctgt atatcgaggt ttatttatta atttgaatag atattaagtt ttattatatt
     1021 tacacttaca tactaataat aaattcaaca aacaatttat ttatgtttat ttatttatta
     1081 aaaaaaaaca aaaactcaaa atttcttcta taaagtaaca aaacttttaa acattctctc
     1141 ttttacaaaa ataaacttat tttgtacttt aaaaacagtc atgttgtatt ataaaataag
     1201 taattagctt aacttataca taatagaaac aaattatact tattagtcag tcagaaacaa
     1261 ctttggcaca tatcaatatt atgctctcga caaataactt ttttgcattt tttgcacgat
     1321 gcatttgcct ttcgccttat tttagagggg cagtaagtac agtaagtacg ttttttcatt
     1381 actggctctt cagtactgtc atctgatgta ccaggcactt catttggcaa aatattagag
     1441 atattatcgc gcaaatatct cttcaaagta ggagcttcta aacgcttacg cataaacgat
     1501 gacgtcaggc tcatgtaaag gtttctcata aattttttgc gactttgaac cttttctccc
     1561 ttgctactga cattatggct gtatataata aaagaattta tgcaggcaat gtttatcatt
     1621 ccgtacaata atgccatagg ccacctattc gtcttcctac tgcaggtcat cacagaacac
     1681 atttggtcta gcgtgtccac tccgccttta gtttgattat aatacataac catttgcggt
     1741 ttaccggtac tttcgttgat agaagcatcc tcatcacaag atgataataa gtataccatc
     1801 ttagctggct tcggtttata tgagacgaga gtaaggggtc cgtcaaaaca aaacatcgat
     1861 gttcccactg gcctggagcg actgtttttc agtacttccg gtatctcgcg tttgtttgat
     1921 cgcacggttc ccacaatggt taattcgagc tcgcccaaac cgggcgcgcc gggcatttaa
     1981 atggaattcg atgccctata tatggatcca attgcaatga tcatcatgac agatctcgac
     2041 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     2101 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2161 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2221 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2281 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2341 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2401 tcatcgctat taccatgggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     2461 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     2521 ggggcggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg
     2581 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
     2641 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
     2701 cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc cgccccggct
     2761 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
     2821 taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg aaagccttaa
     2881 agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg tgcgtgtgtg
     2941 tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc gctgcgggcg
     3001 cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg gggcggtgcc
     3061 ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     3121 ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca cccccctccc
     3181 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
     3241 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
     3301 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
     3361 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
     3421 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
     3481 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     3541 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
     3601 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     3661 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     3721 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgatccgatg gcctcctccg
     3781 aggacgtcat caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc
     3841 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     3901 agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     3961 tccagtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     4021 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     4081 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     4141 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     4201 cctccaccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagatgaggc
     4261 tgaagctgaa ggacggcggc cactacgacg ccgaggtcaa gaccacctac atggccaaga
     4321 agcccgtgca gctgcccggc gcctacaaga ccgacatcaa gctggacatc acctcccaca
     4381 acgaggacta caccatcgtg gaacagtacg agcgcgccga gggccgccac tccaccggcg
     4441 cccccgaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccaa
     4501 cgcgtatgag ctcagagact ggcccagtgg ctgtggaccc cacattgaga cggcggatcg
     4561 agccccatga gtttgaggta ttcttcgatc cgagagagct ccgcaaggag acctgcctgc
     4621 tttacgaaat taattggggg ggccggcact ccatttggcg acatacatca cagaacacta
     4681 acaagcacgt cgaagtcaac ttcatcgaga agttcacgac agaaagatat ttctgtccga
     4741 acacaaggtg cagcattacc tggtttctca gctggagccc atgcggcgaa tgtagtaggg
     4801 ccatcactga attcctgtca aggtatcccc acgtcactct gtttatttac atcgcaaggc
     4861 tgtaccacca cgctgacccc cgcaatcgac aaggcctgcg ggatttgatc tcttcaggtg
     4921 tgactatcca aattatgact gagcaggagt caggatactg ctggagaaac tttgtgaatt
     4981 atagcccgag taatgaagcc cactggccta ggtatcccca tctgtgggta cgactgtacg
     5041 ttcttgaact gtactgcatc atactgggcc tgcctccttg tctcaacatt ctgagaagga
     5101 agcagccaca gctgacattc tttaccatcg ctcttcagtc ttgtcattac cagcgactgc
     5161 ccccacacat tctctgggcc accgggttga aaagcggcag cgagactccc gggacctcag
     5221 agtccgccac acccgaaagt gataaaaagt attctattgg tttagccatc ggcactaatt
     5281 ccgttggatg ggctgtcata accgatgaat acaaagtacc ttcaaagaaa tttaaggtgt
     5341 tggggaacac agaccgtcat tcgattaaaa agaatcttat cggtgccctc ctattcgata
     5401 gtggcgaaac ggcagaggcg actcgcctga aacgaaccgc tcggagaagg tatacacgtc
     5461 gcaagaaccg aatatgttac ttacaagaaa tttttagcaa tgagatggcc aaagttgacg
     5521 attctttctt tcaccgtttg gaagagtcct tccttgtcga agaggacaag aaacatgaac
     5581 ggcaccccat ctttggaaac atagtagatg aggtggcata tcatgaaaag tacccaacga
     5641 tttatcacct cagaaaaaag ctagttgact caactgataa agcggacctg aggttaatct
     5701 acttggctct tgcccatatg ataaagttcc gtgggcactt tctcattgag ggtgatctaa
     5761 atccggacaa ctcggatgtc gacaaactgt tcatccagtt agtacaaacc tataatcagt
     5821 tgtttgaaga gaaccctata aatgcaagtg gcgtggatgc gaaggctatt cttagcgccc
     5881 gcctctctaa atcccgacgg ctagaaaacc tgatcgcaca attacccgga gagaagaaaa
     5941 atgggttgtt cggtaacctt atagcgctct cactaggcct gacaccaaat tttaagtcga
     6001 acttcgactt agctgaagat gccaaattgc agcttagtaa ggacacgtac gatgacgatc
     6061 tcgacaatct actggcacaa attggagatc agtatgcgga cttatttttg gctgccaaaa
     6121 accttagcga tgcaatcctc ctatctgaca tactgagagt taatactgag attaccaagg
     6181 cgccgttatc cgcttcaatg atcaaaaggt acgatgaaca tcaccaagac ttgacacttc
     6241 tcaaggccct agtccgtcag caactgcctg agaaatataa ggaaatattc tttgatcagt
     6301 cgaaaaacgg gtacgcaggt tatattgacg gcggagcgag tcaagaggaa ttctacaagt
     6361 ttatcaaacc catattagag aagatggatg ggacggaaga gttgcttgta aaactcaatc
     6421 gcgaagatct actgcgaaag cagcggactt tcgacaacgg tagcattcca catcaaatcc
     6481 acttaggcga attgcatgct atacttagaa ggcaggagga tttttatccg ttcctcaaag
     6541 acaatcgtga aaagattgag aaaatcctaa cctttcgcat accttactat gtgggacccc
     6601 tggcccgagg gaactctcgg ttcgcatgga tgacaagaaa gtccgaagaa acgattactc
     6661 catggaattt tgaggaagtt gtcgataaag gtgcgtcagc tcaatcgttc atcgagagga
     6721 tgaccaactt tgacaagaat ttaccgaacg aaaaagtatt gcctaagcac agtttacttt
     6781 acgagtattt cacagtgtac aatgaactca cgaaagttaa gtatgtcact gagggcatgc
     6841 gtaaacccgc ctttctaagc ggagaacaga agaaagcaat agtagatctg ttattcaaga
     6901 ccaaccgcaa agtgacagtt aagcaattga aagaggacta ctttaagaaa attgaatgct
     6961 tcgattctgt cgagatctcc ggggtagaag atcgatttaa tgcgtcactt ggtacgtatc
     7021 atgacctcct aaagataatt aaagataagg acttcctgga taacgaagag aatgaagata
     7081 tcttagaaga tatagtgttg actcttaccc tctttgaaga tcgggaaatg attgaggaaa
     7141 gactaaaaac atacgctcac ctgttcgacg ataaggttat gaaacagtta aagaggcgtc
     7201 gctatacggg ctggggacga ttgtcgcgga aacttatcaa cgggataaga gacaagcaaa
     7261 gtggtaaaac tattctcgat tttctaaaga gcgacggctt cgccaatagg aactttatgc
     7321 agctgatcca tgatgactct ttaaccttca aagaggatat acaaaaggca caggtttccg
     7381 gacaagggga ctcattgcac gaacatattg cgaatcttgc tggttcgcca gccatcaaaa
     7441 agggcatact ccagacagtc aaagtagtgg atgagctagt taaggtcatg ggacgtcaca
     7501 aaccggaaaa cattgtaatc gagatggcac gcgaaaatca aacgactcag aaggggcaaa
     7561 aaaacagtcg agagcggatg aagagaatag aagagggtat taaagaactg ggcagccaga
     7621 tcttaaagga gcatcctgtg gaaaataccc aattgcagaa cgagaaactt tacctctatt
     7681 acctacaaaa tggaagggac atgtatgttg atcaggaact ggacataaac cgtttatctg
     7741 attacgacgt cgatcacatt gtaccccaat cctttttgaa ggacgattca atcgacaata
     7801 aagtgcttac acgctcggat aagaaccgag ggaaaagtga caatgttcca agcgaggaag
     7861 tcgtaaagaa aatgaagaac tattggcggc agctcctaaa tgcgaaactg ataacgcaaa
     7921 gaaagttcga taacttaact aaagctgaga ggggtggctt gtctgaactt gacaaggccg
     7981 gatttattaa acgtcagctc gtggaaaccc gccaaatcac aaagcatgtt gcacagatac
     8041 tagattcccg aatgaatacg aaatacgacg agaacgataa gctgattcgg gaagtcaaag
     8101 taatcacttt aaagtcaaaa ttggtgtcgg acttcagaaa ggattttcaa ttctataaag
     8161 ttagggagat aaataactac caccatgcgc acgacgctta tcttaatgcc gtcgtaggga
     8221 ccgcactcat taagaaatac ccgaagctag aaagtgagtt tgtgtatggt gattacaaag
     8281 tttatgacgt ccgtaagatg atcgcgaaaa gcgaacagga gataggcaag gctacagcca
     8341 aatacttctt ttattctaac attatgaatt tctttaagac ggaaatcact ctggcaaacg
     8401 gagagatacg caaacgacct ttaattgaaa ccaatgggga gacaggtgaa atcgtatggg
     8461 ataagggccg ggacttcgcg acggtgagaa aagttttgtc catgccccaa gtcaacatag
     8521 taaagaaaac tgaggtgcag accggagggt tttcaaagga atcgattctt ccaaaaagga
     8581 atagtgataa gctcatcgct cgtaaaaagg actgggaccc gaaaaagtac ggtggcttcg
     8641 atagccctac agttgcctat tctgtcctag tagtggcaaa agttgagaag ggaaaatcca
     8701 agaaactgaa gtcagtcaaa gaattattgg ggataacgat tatggagcgc tcgtcttttg
     8761 aaaagaaccc catcgacttc cttgaggcga aaggttacaa ggaagtaaaa aaggatctca
     8821 taattaaact accaaagtat agtctgtttg agttagaaaa tggccgaaaa cggatgttgg
     8881 ctagcgccgg agagcttcaa aaggggaacg aactcgcact accgtctaaa tacgtgaatt
     8941 tcctgtattt agcgtcccat tacgagaagt tgaaaggttc acctgaagat aacgaacaga
     9001 agcaactttt tgttgagcag cacaaacatt atctcgacga aatcatagag caaatttcgg
     9061 aattcagtaa gagagtcatc ctagctgatg ccaatctgga caaagtatta agcgcataca
     9121 acaagcacag ggataaaccc atacgtgagc aggcggaaaa tattatccat ttgtttactc
     9181 ttaccaacct cggcgctcca gccgcattca agtattttga cacaacgata gatcgcaaac
     9241 gatacacttc taccaaggag gtgctagacg cgacactgat tcaccaatcc atcacgggat
     9301 tatatgaaac tcggatagat ttgtcacagc ttgggggtga ctctggtggt tctactaatc
     9361 tgtcagatat tattgaaaag gagaccggta agcaactggt tatccaggaa tccatcctca
     9421 tgctcccaga ggaggtggaa gaagtcattg ggaacaagcc ggaaagcgat atactcgtgc
     9481 acaccgccta cgacgagagc accgacgaga atgtcatgct tctgactagc gacgcccctg
     9541 aatacaagcc ttgggctctg gtcatacagg atagcaacgg tgagaacaag attaagatgc
     9601 tctctggtgg ttctcccaag aagaagagga aagtctaacc ggtcatcatc accatcacca
     9661 ttgagtttaa acccgctgat cagcacgcgt gctagcttcg aagctaattc actcctcagg
     9721 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     9781 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     9841 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     9901 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     9961 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
    10021 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
    10081 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
    10141 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
    10201 cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg
    10261 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    10321 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    10381 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
    10441 gatctgacaa tgttcagtgc agagactcgg ctacgcctcg tggactttga agttgaccaa
    10501 caatgtttat tcttacctct aatagtcctc tgtggcaagg tcaagattct gttagaagcc
    10561 aatgaagaac ctggttgttc aataacattt tgttcgtcta atatttcact accgcttgac
    10621 gttggctgca cttcatgtac ctcatctata aacgcttctt ctgtatcgct ctggacgtca
    10681 tcttcactta cgtgatctga tatttcactg tcagaatcct caccaacaag ctcgtcatcg
    10741 ctttgcagaa gagcagagag gatatgctca tcgtctaaag aactacccat tttattatat
    10801 attagtcacg atatctataa caagaaaata tatatataat aagttatcac gtaagtagaa
    10861 catgaaataa caatataatt atcgtatgag ttaaatctta aaagtcacgt aaaagataat
    10921 catgcgtcat tttgactcac gcggtcgtta tagttcaaaa tcagtgacac ttaccgcatt
    10981 gacaagcacg cctcacggga gctccaagcg gcgactgaga tgtcctaaat gcacagcgac
    11041 ggattcgcgc tatttagaaa gagagagcaa tatttcaaga atgcatgcgt caattttacg
    11101 cagactatct ttctagggtt aaaaaagatt tgcgctttac tcgacctaaa ctttaaacac
    11161 gtcatagaat cttcgtttga caaaaaccac attgtggcca agctgtgtga cgcgacgcgc
    11221 gctaaagaat ggcaaaccaa gtcgcgcgag cgtcgacctc gagggggggc ccggtaccca
    11281 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
    11341 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
    11401 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
    11461 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
    11521 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
    11581 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
    11641 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
    11701 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
    11761 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
    11821 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
    11881 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
    11941 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
    12001 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
    12061 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
    12121 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
    12181 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
    12241 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
    12301 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
    12361 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
    12421 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
    12481 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
    12541 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
    12601 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
    12661 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
    12721 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
    12781 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
    12841 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
    12901 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
    12961 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
    13021 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
    13081 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
    13141 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    13201 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    13261 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    13321 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    13381 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
    13441 aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
