LOCUS       sequence-492311-       12139 bp    DNA     circular SYN 07-NOV-2025
DEFINITION  Expresses BirA*-dCas9-eGFP in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12139)
  AUTHORS   Schmidtmann E, Anton T, Rombaut P, Herzog F, Leonhardt H
  TITLE     Determination of Local Chromatin Composition by CasID.
  JOURNAL   Nucleus. 2016 Sep 27:0.
  PUBMED    27676121
REFERENCE   2  (bases 1 to 12139)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleus. 
            2016 Sep 27:0."
COMMENT     Sequence Label: pCAG-BirA*-dCas9-eGFP
FEATURES             Location/Qualifiers
     source          1..12139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        69..372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        374..652
                     /label=chicken beta-actin promoter
     primer_bind     1593..1615
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1626..1655
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(1659..2006,2007..2009,2010..2618)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             join(2733..2759,2760..2762,2763..5246,5247..5252,5253..5318,
                     5319..5321,5322..6836)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(2733..2759,2760..2762,2763..5249,5250..5252,
                     5253..6836)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2733..6836
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6849..6869
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      6925..6934
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6945..6965
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(6966..6968,6969..6971,6972..7685)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6966..6968,6969..6971,6972..7682)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7011..7032)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7272..7291)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7619..7640
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7724..8298
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7892..7909)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8119..8138
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      8303..8312
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8309..8707
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    8865..8986
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8902..8921)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8956..8975
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9034..9169
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9275..9292)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9446..10034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9526..9545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10205..10996,10997..11065))
                     /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     10828..10847
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11066..11170)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     11238..11256
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(11294..11316)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     11416..11435
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(11502..11957)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11639..11660)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11851..11870
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12083..12105
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12097..12114
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12098..12114
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        12121..12139
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 acgcgtccta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
       61 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      121 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      181 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      241 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      301 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      361 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
      421 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
      481 gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc
      541 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
      601 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
      661 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
      721 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      781 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga
      841 ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt
      901 gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg
      961 cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc
     1021 cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg
     1081 ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc
     1141 ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg
     1201 gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc
     1261 tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg
     1321 aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact
     1381 tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag
     1441 cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg
     1501 tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga
     1561 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     1621 ctatggaaca aaaactcatc tcagaagagg atctcgacaa ggacaacacc gtgcccctga
     1681 agctgatcgc cctgctggcc aacggcgagt tccactctgg cgagcagctg ggagagaccc
     1741 tgggaatgag cagagccgcc atcaacaagc acatccagac actgagagac tggggagtgg
     1801 acgtgttcac cgtgcctggc aagggctaca gcctgcctga gcctatccag ctgctgaacg
     1861 ccaagcagat cctgggacag ctggatggcg gaagcgtggc cgtgctgcct gtgatcgact
     1921 ccaccaatca gtacctgctg gacagaatcg gagagctgaa gtccggcgac gcctgcatcg
     1981 ccgagtacca gcaggctggc agaggaggca gaggacggaa gtggttcagc ccattcggag
     2041 ccaacctgta cctgtccatg ttctggagac tggagcaggg acctgctgct gccatcggac
     2101 tgagtctggt gatcggaatc gtgatggccg aggtgctgag aaagctggga gccgacaagg
     2161 tgagagtgaa gtggcctaat gacctgtacc tccaggaccg caagctggct ggcatcctgg
     2221 tggagctgac aggcaagaca ggcgatgccg ctcagatcgt gatcggagcc ggaatcaaca
     2281 tggccatgag aagagtggag gagagcgtgg tgaaccaggg ctggatcacc ctgcaggagg
     2341 ctggcatcaa cctggaccgg aacaccctgg ccgccatgct gatcagagag ctgagagccg
     2401 ctctggagct gttcgagcag gagggactgg ctccttacct gagcagatgg gagaagctgg
     2461 acaacttcat caacagacct gtgaagctga tcatcggcga caaggaaatc ttcggcatct
     2521 ccagaggaat cgacaagcag ggagctctgc tgctggagca ggacggaatc atcaagccct
     2581 ggatgggcgg agaaatctcc ctgagaagcg cagagaagct cgagcggccg ccactgtgct
     2641 ggatatctgc agaattccac cacactggac tagtggatcc gagctcggta ccaagcttaa
     2701 gtttaaaccg cggtagcggc agcggtagcg gaatggacaa gaagtactcc attgggctcg
     2761 ctatcggcac aaacagcgtc ggctgggccg tcattacgga cgagtacaag gtgccgagca
     2821 aaaaattcaa agttctgggc aataccgatc gccacagcat aaagaagaac ctcattggcg
     2881 ccctcctgtt cgactccggg gagacggccg aagccacgcg gctcaaaaga acagcacggc
     2941 gcagatatac ccgcagaaag aatcggatct gctacctgca ggagatcttt agtaatgaga
     3001 tggctaaggt ggatgactct ttcttccata ggctggagga gtcctttttg gtggaggagg
     3061 ataaaaagca cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg gcgtaccatg
     3121 aaaagtaccc aaccatatat catctgagga agaagcttgt agacagtact gataaggctg
     3181 acttgcggtt gatctatctc gcgctggcgc atatgatcaa atttcgggga cacttcctca
     3241 tcgaggggga cctgaaccca gacaacagcg atgtcgacaa actctttatc caactggttc
     3301 agacttacaa tcagcttttc gaagagaacc cgatcaacgc atccggagtt gacgccaaag
     3361 caatcctgag cgctaggctg tccaaatccc ggcggctcga aaacctcatc gcacagctcc
     3421 ctggggagaa gaagaacggc ctgtttggta atcttatcgc cctgtcactc gggctgaccc
     3481 ccaactttaa atctaacttc gacctggccg aagatgccaa gcttcaactg agcaaagaca
     3541 cctacgatga tgatctcgac aatctgctgg cccagatcgg cgaccagtac gcagaccttt
     3601 ttttggcggc aaagaacctg tcagacgcca ttctgctgag tgatattctg cgagtgaaca
     3661 cggagatcac caaagctccg ctgagcgcta gtatgatcaa gcgctatgat gagcaccacc
     3721 aagacttgac tttgctgaag gcccttgtca gacagcaact gcctgagaag tacaaggaaa
     3781 ttttcttcga tcagtctaaa aatggctacg ccggatacat tgacggcgga gcaagccagg
     3841 aggaatttta caaatttatt aagcccatct tggaaaaaat ggacggcacc gaggagctgc
     3901 tggtaaagct taacagagaa gatctgttgc gcaaacagcg cactttcgac aatggaagca
     3961 tcccccacca gattcacctg ggcgaactgc acgctatcct caggcggcaa gaggatttct
     4021 accccttttt gaaagataac agggaaaaga ttgagaaaat cctcacattt cggataccct
     4081 actatgtagg ccccctcgcc cggggaaatt ccagattcgc gtggatgact cgcaaatcag
     4141 aagagaccat cactccctgg aacttcgagg aagtcgtgga taagggggcc tctgcccagt
     4201 ccttcatcga aaggatgact aactttgata aaaatctgcc taacgaaaag gtgcttccta
     4261 aacactctct gctgtacgag tacttcacag tttataacga gctcaccaag gtcaaatacg
     4321 tcacagaagg gatgagaaag ccagcattcc tgtctggaga gcagaagaaa gctatcgtgg
     4381 acctcctctt caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa gactatttca
     4441 aaaagattga atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc ttcaacgcat
     4501 ccctgggaac gtatcacgat ctcctgaaaa tcattaaaga caaggacttc ctggacaatg
     4561 aggagaacga ggacattctt gaggacattg tcctcaccct tacgttgttt gaagataggg
     4621 agatgattga agaacgcttg aaaacttacg ctcatctctt cgacgacaaa gtcatgaaac
     4681 agctcaagag gcgccgatat acaggatggg ggcggctgtc aagaaaactg atcaatggga
     4741 tccgagacaa gcagagtgga aagacaatcc tggattttct taagtccgat ggatttgcca
     4801 acaggaactt catgcagttg atccatgatg actctctcac ctttaaggag gacatccaga
     4861 aagcacaagt ttctggccag ggggacagtc ttcacgagca catcgctaat cttgcaggta
     4921 gcccagctat caaaaaggga atactgcaga ccgttaaggt cgtggatgaa ctcgtcaaag
     4981 taatgggaag gcataagccc gagaatatcg ttatcgagat ggcccgagag aaccaaacta
     5041 cccagaaggg acagaagaac agtagggaaa ggatgaagag gattgaagag ggtataaaag
     5101 aactggggtc ccaaatcctt aaggaacacc cagttgaaaa cacccagctt cagaatgaga
     5161 agctctacct gtactacctg cagaacggca gggacatgta cgtggatcag gaactggaca
     5221 tcaatcggct ctccgactac gacgtggatg ctatcgtgcc ccagtctttt ctcaaagatg
     5281 attctattga taataaagtg ttgacaagat ccgataaaaa tagagggaag agtgataacg
     5341 tcccctcaga agaagttgtc aagaaaatga aaaattattg gcggcagctg ctgaacgcca
     5401 aactgatcac acaacggaag ttcgataatc tgactaaggc tgaacgaggt ggcctgtctg
     5461 agttggataa agccggcttc atcaaaaggc agcttgttga gacacgccag atcaccaagc
     5521 acgtggccca aattctcgat tcacgcatga acaccaagta cgatgaaaat gacaaactga
     5581 ttcgagaggt gaaagttatt actctgaagt ctaagctggt ctcagatttc agaaaggact
     5641 ttcagtttta taaggtgaga gagatcaaca attaccacca tgcgcatgat gcctacctga
     5701 atgcagtggt aggcactgca cttatcaaaa aatatcccaa gcttgaatct gaatttgttt
     5761 acggagacta taaagtgtac gatgttagga aaatgatcgc aaagtctgag caggaaatag
     5821 gcaaggccac cgctaagtac ttcttttaca gcaatattat gaattttttc aagaccgaga
     5881 ttacactggc caatggagag attcggaagc gaccacttat cgaaacaaac ggagaaacag
     5941 gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc ctgtccatgc
     6001 cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg aggcttctcc aaggaaagta
     6061 tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa aaaagattgg gaccccaaga
     6121 aatacggcgg attcgattct cctacagtcg cttacagtgt actggttgtg gccaaagtgg
     6181 agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc acaatcatgg
     6241 agcgatcaag cttcgaaaaa aaccccatcg actttctcga ggcgaaagga tataaagagg
     6301 tcaaaaaaga cctcatcatt aagcttccca agtactctct ctttgagctt gaaaacggcc
     6361 ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg taacgagctg gcactgccct
     6421 ctaaatacgt taatttcttg tatctggcca gccactatga aaagctcaaa gggtctcccg
     6481 aagataatga gcagaagcag ctgttcgtgg aacaacacaa acactacctt gatgagatca
     6541 tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc cgacgctaac ctcgataagg
     6601 tgctttctgc ttacaataag cacagggata agcccatcag ggagcaggca gaaaacatta
     6661 tccacttgtt tactctgacc aacttgggcg cgcctgcagc cttcaagtac ttcgacacca
     6721 ccatagacag aaagcggtac acctctacaa aggaggtcct ggacgccaca ctgattcatc
     6781 agtcaattac ggggctctat gaaacaagaa tcgacctctc tcagctcggt ggagacagca
     6841 gggctgaccc caagaagaag aggaaggtga ttgtacaaga gcggcctgag gagcagagcc
     6901 caggcgagca acagcgccgt ggacgccacc atgggcgatc gcatccaaag aagaagagaa
     6961 aggtcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     7021 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     7081 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     7141 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     7201 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     7261 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     7321 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     7381 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     7441 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     7501 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     7561 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     7621 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     7681 agtaagcggc cgcagttaac gaattccgcc cccccccccc ccccccctct ccctcccccc
     7741 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     7801 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     7861 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     7921 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     7981 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     8041 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     8101 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     8161 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     8221 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     8281 tgaaaaacac gatgataagc ttgccaccat ggccaagcct ttgtctcaag aagaatccac
     8341 cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt
     8401 cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt
     8461 tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg
     8521 caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg
     8581 acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag
     8641 tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga
     8701 gggctaagca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
     8761 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     8821 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     8881 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     8941 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     9001 cgtcgacctc gaggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cttcactact
     9061 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     9121 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     9181 ggggcgggac tatggttgct gactaattgc tgcattaatg aatcggccaa cgcgcgggga
     9241 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     9301 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     9361 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     9421 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     9481 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     9541 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     9601 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     9661 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     9721 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     9781 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     9841 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     9901 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     9961 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
    10021 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
    10081 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
    10141 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
    10201 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
    10261 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
    10321 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
    10381 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
    10441 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
    10501 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
    10561 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
    10621 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
    10681 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
    10741 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
    10801 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
    10861 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
    10921 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
    10981 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
    11041 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
    11101 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
    11161 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
    11221 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
    11281 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
    11341 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
    11401 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
    11461 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
    11521 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
    11581 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
    11641 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
    11701 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
    11761 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
    11821 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
    11881 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgctcaatgc
    11941 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
    12001 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
    12061 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg
    12121 taatacgact cactatagg
//
