LOCUS       sequence-462114-       15483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the PE2 prime-editing machinery fused to BFP 
            (PE2-P2A-BFP), under the control of a TRE3G promoter. This promoter 
            is responsive to doxycycline bound to the rtTA protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15483)
  AUTHORS   Martyn GE, Montgomery MT, Jones H, Guo K, Doughty BR, Linder J, Chen
            Z, Cochran K, Lawrence KA, Munson G, Pampari A, Fulco CP, Kelley DR,
            Lander ES, Kundaje A, Engreitz JM
  TITLE     Rewriting regulatory DNA to dissect and reprogram gene expression.
  JOURNAL   bioRxiv [Preprint]. 2023 Dec 21:2023.12.20.572268. doi: 
            10.1101/2023.12.20.572268.
  PUBMED    38187584
REFERENCE   2  (bases 1 to 15483)
  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: "bioRxiv 
            [Preprint]. 2023 Dec 21:2023.12.20.572268. doi: 
            10.1101/2023.12.20.572268."
COMMENT     Sequence Label: TRE3G-PE2-P2A-BFP
FEATURES             Location/Qualifiers
     source          1..15483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1040..1161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1105..1124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1291..1665)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(1684..1731)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1779..2108)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1793..1928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1847..1866)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2093..2113
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2122..2550)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2232..2253)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2444..2463
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2596..2820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2809..2826
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             2852..3032
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     3537..3553
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3538..3554
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3557..4145
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3663..3674
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4072..4092
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4189..4887)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             complement(4888..4944)
                     /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             complement(4954..4974)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(7147..8730,8731..8733,8734..11247))
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(7147..11247)
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(11248..11268)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(11341..11690)
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     primer_bind     complement(11378..11402)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    11448..11466
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11484..11502
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11520..11538
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11556..11574
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11592..11610
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11628..11646
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11664..11682
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    11763..11880
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(12178..12222)
                     /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    12407..12640
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    13133..13258
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             13305..13485
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(13503..13701)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        13703..14082
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     14254..14273
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        14348..14452
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(14453..14521,14522..15313)
                     /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(14671..14690)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1081 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1141 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1201 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1261 ggcggtggaa tcgaaatctc gtagcacgtg tcagtcctgc tcctcggcca cgaagtgcac
     1321 gcagttgccg gccgggtcgc gcagggcgaa ctcccgcccc cacggctgct cgccgatctc
     1381 ggtcatggcc ggcccggagg cgtcccggaa gttcgtggac acgacctccg accactcggc
     1441 gtacagctcg tccaggccgc gcacccacac ccaggccagg gtgttgtccg gcaccacctg
     1501 gtcctggacc gcgctgatga acagggtcac gtcgtcccgg accacaccgg cgaagtcgtc
     1561 ctccacgaag tcccgggaga acccgagccg gtcggtccag aactcgaccg ctccggcgac
     1621 gtcgcgcgcg gtgagcaccg gaacggcact ggtcaacttg gccatggttt agttcctcac
     1681 cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa cacgtgctga
     1741 tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca
     1801 aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc ggcctctgca
     1861 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     1921 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     1981 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     2041 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     2101 tgacacacat tccacagaat taattcgcgt taaatttttg ttaaatcagc tcatttttta
     2161 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     2221 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     2281 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     2341 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     2401 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     2461 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     2521 ccgcgcttaa tgcgccgcta cagggcgcgt ggggataccc cctagagccc cagctggttc
     2581 tttccgcctc agaagccata gagcccaccg catccccagc atgcctgcta ttgtcttccc
     2641 aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca
     2701 gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca
     2761 gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag
     2821 tcgaggctga tcagcgggtt taaacgggcc ctgctagaga ttttccacac tgactaaaag
     2881 ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc
     2941 actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc
     3001 caggctcaga tctggtctaa ccagagagac ccagtacagt ccggatgcag ctctcgggcc
     3061 atgtgatgaa atgctaggcg gctgtcaaac ctccactcta atacttctct ctccgggtca
     3121 tccatcccat gcaggctcac agggtgtaac aagcgggtgt tctctccttc attggcttct
     3181 tctaccttct cttgctcaac tggtactagc ttgtagcacc atccaaaggt cagtggatat
     3241 ctgatccctg gccctggtgt gtagttctgc caatcaggga agtagccttg tgtgtggtag
     3301 atccacagat caaggatatc ttgtcttcgt tgggagtgaa ttagcccttc cagtcccccc
     3361 ttttctttta aaaagtggct aagatctaca gctgccttgt aagtcattgg tcttaaaggt
     3421 acctgaggtg tgactggaaa acccacctcc tcctcctctt gtgcttctag ccaggcacaa
     3481 tcagcattgg tagctgctgt attgctactt gtgattgctc catgtttttc taggtctcga
     3541 ggtcgacggt atcgatgcgg ggaggcggcc caaagggaga tccgactcgt ctgagggcga
     3601 aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct
     3661 ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca
     3721 caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga attgtcagtg
     3781 cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg
     3841 gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc
     3901 caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg cctgacaacg
     3961 ggccacaact cctcataaag agacagcaac caggatttat acaaggagga gaaaatgaaa
     4021 gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta
     4081 aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta atccagaggt
     4141 tgattatcga taagcttgat atcaagcttg catgcctgca ggactagttt aattaagctt
     4201 gtgccccagt ttgctaggga ggtcgcagta tctggccact gccacctcgt gctgctcgac
     4261 gtaggtctcg ttgttggcct ccttgattct ttccagtctg tagtccacat agtagacgcc
     4321 aggcatcttg aggttcttag cgggtttctt ggatctatat gtggtcttga tgtttgcgat
     4381 cagatggctc ccgcccacga gcttcagggc catgtcgttt ctgccttcca ggccgccgtc
     4441 agcggggtac agcgtctcgg tgaaggcctc ccagccgagt gttttcttct gcatcacagg
     4501 gccgttggat gtgaagttca cccctctgat cttgacgttg tagatgaggc agccgtcctg
     4561 gaggctggtg tcctgggtag cggtcagcac gcccccgtct tcgtatgtgg tgactctctc
     4621 ccatgtgaag ccctcaggga aggactgctt gaagaagtcg gggatgccct gggtgtggtt
     4681 gatgaaggtc ttgctgccgt agaggaagct agtagccagg atgtcgaagg cgaaggggag
     4741 agggccgccc tcgaccacct tgattctcat ggtctgggtg ccctcgtagg gcttgccttc
     4801 gccctcggat gtgcacttga agtgatggtt gtccacggtg ccctccatgt acagcttcat
     4861 gtgcatgttc tccttaatca gctcgctcgg tccaggattc tcttcgacat ctccggcttg
     4921 tttcagcaga gagaagtttg ttgcgccgga tccgactttc ctcttcttct tgggctcgaa
     4981 ttcgctgccg tcggcggttc tttttgagcc gccagagggt gatgaatttt ctatgaggag
     5041 ggtagaggtg tctggagtct ctgtgatggc tgcctttcgg gccgcttggt cagccatccg
     5101 gttgcctcta gcctcggcgc tgtgtccctt ttgatgtcct ggacaatgga ttatgctaag
     5161 tcttttgggc agaaagaggg cttttagtag ggccaagatc tcgtctttat ttttgatctc
     5221 tttgccttct gatgtgagcc acccacgcct tctgtatatt tctccatgga tatgggcagt
     5281 agcaaaagca taacggctat cagtataaac atttagcttc ttaccttctg ccatctttag
     5341 ggcctgggtg agtgctatca gttcagcccg ctgagcggat gtcccggctg gcagggcttt
     5401 agcccagatt acctcggtct cggtggtcac cgcagctccc gccttacgct gtccctcttg
     5461 taagagactg cttccatccg tgtaccaggt gtggtcggcg tctgggagcg gctggtccgt
     5521 taggtcgggt cgggttccgt gggcttcggc caggatatca aggcagttgt gttgcagccc
     5581 ttcctcaggc agtgggagca gcgtagccgg gttcagggct accaccggtc cgaactggac
     5641 ccggtccgtg tccaaaagca aggcctgata gtgagtcatc cgggcgttgg aaagccagcg
     5701 gtcggggggt tgtttgacta gtgcctctac tgcatggggg gccagaatga ctagtggctg
     5761 tcccatggtt agcttgcctg catcctttgt cagtacggca atggctgcta ccatccgtag
     5821 gcaagggggc cacccagctg ctactgggtc tagctttttg gacaggtagg ccaccggccg
     5881 acgccaaggt cccagttttt gcgttaggac acctttggcg tagccctgct tctcgtcgac
     5941 aaagagttca aagggcttag tcaaatctgg caaccccagg gctggggcag ttagaagagc
     6001 ttgcttgatt tcttgatagg ccttttgttg gtctgggccc caattaaaca gagtccccgg
     6061 tttggtgaga gggtacaggg gggctgccat ttctgcaaac ccagggatga agaggcgaca
     6121 gaagcctgcc ttccctagga actcccttag ttgtcgaggg gtcttcggag taggctgccc
     6181 catcacagtc tcttttctgg cctcagtcag ccatctctga ccctctttta gaagataccc
     6241 cagatacttg acctgtttct ggcaaatttg ggctttcttg gccgaggccc gatacccgag
     6301 gttccctagg gtttgtaaca gggcccgagt accttgttgg cagtctagct cagaagtggc
     6361 ggccagcagt aagtcatcca cgtactgtag caggatcaag tctgggtgct ggatccggaa
     6421 gtctgctagg tctctgtgca gtgcctcatt aaacagggtg ggactgtttt tgaaaccctg
     6481 tgggagtctg gtccaggtca attgtcctga gattcccatc tctggatctc tccactcaaa
     6541 ggcgaagaga ggctgactgg tggggtggag tctcaggcag aaaaaggcat cctttaaatc
     6601 aagcacagtg taccactggt gggacggtgg gagcccgctc aagaggttgt aagggttggg
     6661 cacggtgggg tggatgtctt ccacccgctt gttgacttct ctcagatcct ggacaggcct
     6721 ataatcatta gtccctggtt tcttaacggg tagcaggggc gtgttccagg gggactggca
     6781 gggtaccagt attccctggt ccaacagtct ctgtatgtgg ggcttgatcc ccagtctggc
     6841 ttcttgtgac atggggtatt gttttatgga cacgggggta gaggttgctt tcagaggtat
     6901 gatcagagga gcttggcgaa ctgccagtcc catgcccccg gtttccgccc aggcctgagg
     6961 aaaatcagac agccatgtgg accctagaga aacatctggc tcttttgagg tctcatgtag
     7021 ccgatactca tcttctatat ttagggtgct gctgccgccg ctgctgccgc cactgctctc
     7081 tggtgttgct gactcgcttg ttcctggtgt ctcgctgcca gaggatcctc cgctagatcc
     7141 tccagagtca cctcccagct gagacaggtc gatccgtgtc tcgtacaggc cggtgatgct
     7201 ctggtggatc agggtggcgt ccagcacctc tttggtgctg gtgtacctct tccggtcgat
     7261 ggtggtgtca aagtacttga aggcggcagg ggctcccaga ttggtcaggg taaacaggtg
     7321 gatgatattc tcggcctgct ctctgatggg cttatcccgg tgcttgttgt aggcggacag
     7381 cactttgtcc agattagcgt cggccaggat cactctcttg gagaactcgc tgatctgctc
     7441 gatgatctcg tccaggtagt gcttgtgctg ttccacaaac agctgtttct gctcattatc
     7501 ctcgggggag cccttcagct tctcatagtg gctggccagg tacaggaagt tcacatattt
     7561 ggagggcagg gccagttcgt ttcccttctg cagttcgccg gcagaggcca gcattctctt
     7621 ccggccgttt tccagctcga acagggagta cttaggcagc ttgatgatca ggtccttttt
     7681 cacttctttg tagcccttgg cttccagaaa gtcgatggga ttcttctcga agctgcttct
     7741 ttccatgatg gtgatcccca gcagctcttt cacactcttc agtttcttgg acttgccctt
     7801 ttccactttg gccaccacca gcacagaata ggccacggtg gggctgtcga agccgccgta
     7861 cttcttaggg tcccagtcct tctttctggc gatcagctta tcgctgttcc tcttgggcag
     7921 gatagactct ttgctgaagc cgcctgtctg cacctcggtc tttttcacga tattcacttg
     7981 gggcatgctc agcactttcc gcacggtggc aaaatcccgg cccttatccc acacgatctc
     8041 cccggtttcg ccgtttgtct cgatcagagg ccgcttccgg atctcgccgt tggccagggt
     8101 aatctcggtc ttgaaaaagt tcatgatgtt gctgtagaag aagtacttgg cggtagcctt
     8161 gccgatttcc tgctcgctct tggcgatcat cttccgcacg tcgtacacct tgtagtcgcc
     8221 gtacacgaac tcgctttcca gcttagggta ctttttgatc agggcggttc ccacgacggc
     8281 gttcaggtag gcgtcgtggg cgtggtggta gttgttgatc tcgcgcactt tgtaaaactg
     8341 gaaatccttc cggaaatcgg acaccagctt ggacttcagg gtgatcactt tcacttcccg
     8401 gatcagcttg tcattctcgt cgtacttagt gttcatccgg gagtccagga tctgtgccac
     8461 gtgctttgtg atctgccggg tttccaccag ctgtctcttg atgaagccgg ccttatccag
     8521 ttcgctcagg ccgcctctct cggccttggt cagattgtcg aactttctct gggtaatcag
     8581 cttggcgttc agcagctgcc gccagtagtt cttcatcttc ttcacgacct cttcggaggg
     8641 cacgttgtcg ctcttgcccc ggttcttgtc gcttctggtc agcaccttgt tgtcgatgga
     8701 gtcgtccttc agaaagctct gaggcacgat agcgtccaca tcgtagtcgg acagccggtt
     8761 gatgtccagt tcctggtcca cgtacatatc ccgcccattc tgcaggtagt acaggtacag
     8821 cttctcgttc tgcagctggg tgttttccac ggggtgttct ttcaggatct ggctgcccag
     8881 ctctttgatg ccctcttcga tccgcttcat tctctcgcgg ctgttcttct gtcccttctg
     8941 ggtggtctgg ttctctctgg ccatttcgat cacgatgttc tcgggcttgt gccggcccat
     9001 cactttcacg agctcgtcca ccaccttcac tgtctgcagg atgcccttct taatggcggg
     9061 gctgccggcc agattggcaa tgtgctcgtg caggctatcg ccctggccgg acacctgggc
     9121 tttctggatg tcctctttaa aggtcaggct gtcgtcgtgg atcagctgca tgaagtttct
     9181 gttggcgaag ccgtcggact tcaggaaatc caggattgtc ttgccggact gcttgtcccg
     9241 gatgccgttg atcagcttcc ggctcagcct gccccagccg gtgtatctcc gccgcttcag
     9301 ctgcttcatc actttgtcgt cgaacaggtg ggcataggtt ttcagccgtt cctcgatcat
     9361 ctctctgtcc tcaaacagtg tcagggtcag cacgatatct tccagaatgt cctcgttttc
     9421 ctcattgtcc aggaagtcct tgtccttgat aattttcagc agatcgtggt atgtgcccag
     9481 ggaggcgttg aaccgatctt ccacgccgga gatttccacg gagtcgaagc actcgatttt
     9541 cttgaagtag tcctctttca gctgcttcac ggtcactttc cggttggtct tgaacagcag
     9601 gtccacgatg gcctttttct gctcgccgct caggaaggcg ggctttctca ttccctcggt
     9661 cacgtatttc actttggtca gctcgttata cacggtgaag tactcgtaca gcaggctgtg
     9721 cttgggcagc accttctcgt tgggcaggtt cttatcgaag ttggtcatcc gctcgatgaa
     9781 gctctgggcg gaagcgccct tgtccaccac ttcctcgaag ttccaggggg tgatggtttc
     9841 ctcgctcttt ctggtcatcc aggcgaatct gctgtttccc ctggccagag ggcccacgta
     9901 gtaggggatg cggaaggtca ggatcttctc gatcttttcc cggttgtcct tcaggaatgg
     9961 gtaaaaatct tcctgccgcc gcagaatggc gtgcagctct cccaggtgga tctggtgggg
    10021 gatgctgccg ttgtcgaagg tccgctgctt ccgcagcagg tcctctctgt tcagcttcac
    10081 gagcagttcc tcggtgccgt ccatcttttc caggatgggc ttgatgaact tgtagaactc
    10141 ttcctggctg gctccgccgt caatgtagcc ggcgtagccg ttcttgctct ggtcgaagaa
    10201 aatctctttg tacttctcag gcagctgctg ccgcacgaga gctttcagca gggtcaggtc
    10261 ctggtggtgc tcgtcgtatc tcttgatcat agaggcgctc aggggggcct tggtgatctc
    10321 ggtgttcact ctcaggatgt cgctcagcag gatggcgtcg gacaggttct tggcggccag
    10381 aaacaggtcg gcgtactggt cgccgatctg ggccagcagg ttgtccaggt cgtcgtcgta
    10441 ggtgtccttg ctcagctgca gtttggcatc ctcggccagg tcgaagttgc tcttgaagtt
    10501 gggggtcagg cccaggctca gggcaatcag gtttccgaac aggccattct tcttctcgcc
    10561 gggcagctgg gcgatcagat tttccagccg tctgctcttg ctcagtctgg cagacaggat
    10621 ggccttggcg tccacgccgc tggcgttgat ggggttttcc tcgaacagct ggttgtaggt
    10681 ctgcaccagc tggatgaaca gcttgtccac gtcgctgttg tcggggttca ggtcgccctc
    10741 gatcaggaag tggccccgga acttgatcat gtgggccagg gccagataga tcagccgcag
    10801 gtcggccttg tcggtgctgt ccaccagttt ctttctcagg tggtagatgg tggggtactt
    10861 ctcgtggtag gccacctcgt ccacgatgtt gccgaagatg gggtgccgct cgtgcttctt
    10921 atcctcttcc accaggaagg actcttccag tctgtggaag aagctgtcgt ccaccttggc
    10981 catctcgttg ctgaagatct cttgcagata gcagatccgg ttcttccgtc tggtgtatct
    11041 tcttctggcg gttctcttca gccgggtggc ctcggctgtt tcgccgctgt cgaacagcag
    11101 ggctccgatc aggttcttct tgatgctgtg ccggtcggtg ttgcccagca ccttgaattt
    11161 cttgctgggc accttgtact cgtcggtgat cacggcccag cccacagagt tggtgccgat
    11221 gtccaggccg atgctgtact tcttgtcgac tttccgcttc ttctttggtg actcgaactc
    11281 gcttccgtcg gctgtccgtt tcatggtggc accggtaagc ttgatatcct gcagacgcgt
    11341 aagtataaga caaaagtgtt gtggaattgc tccaggcgat ctgacggttc actaaacgag
    11401 ctctgctttt ataggcgccc accgtacacg cctacctcga catacgttct ctatcactga
    11461 tagggagtaa actggatata cgttctctat cactgatagg gagtaaactg tagatacgtt
    11521 ctctatcact gatagggagt aaactggtca tacgttctct atcactgata gggagtaaac
    11581 tccttatacg ttctctatca ctgataggga gtaaagtctg catacgttct ctatcactga
    11641 tagggagtaa actcttcata cgttctctat cactgatagg gagtaaactc gagtgaagac
    11701 gaaagggcct ctagacactt aattaaccaa actggatctc tgctgtccct gtaataaacc
    11761 cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct
    11821 attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa
    11881 ttgtggatga atactgccat ttgtctgcag aattggcgca cgcagtgccg atccgttcac
    11941 taatcgaatg gatctgtctc tgtctctctc tccaccttct tcttctattc cttcgggcct
    12001 gtcgggtccc ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct
    12061 aactctattc actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac
    12121 tatcattatg aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa
    12181 acttgcccat ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga
    12241 ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat
    12301 ccaggtcgtg tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg
    12361 cacagcagtg gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc
    12421 ctttaggtat ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt
    12481 gagttgcaac agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac
    12541 tataccagac aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt
    12601 gcttcctgct gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc
    12661 tctctgcacc actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac
    12721 tactttatat ttatataatt cacttctcca attgtccctc atatctcctc ctccaggtct
    12781 gaagatcagc ggccgcttgc tgtgcggtgg tcttactttt gttttgctct tcctctatct
    12841 tgtctaaagc ttccttggtg tcttttatct ctatcctttg atgcacacaa tagagggttg
    12901 ctactgtatt atataatgat ctaagttctt ctgatcctgt ctgaagggat ggttgtagct
    12961 gtcccagtat ttgtctacag ccttctgatg tttctaacag gccaggatta actgcgaatc
    13021 gttctagctc cctgcttgcc catactatat gttttaattt atattttttc tttccccctg
    13081 gccttaaccg aattttttcc catcgcgatc taattctccc ccgcttaata ctgacgctct
    13141 cgcacccatc tctctccttc tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc
    13201 gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct
    13261 cctctggttt ccctttcgct ttcaagtccc tgttcgggcg ccactgctag agattttcca
    13321 cactgactaa aagggtctga gggatctcta gttaccagag tcacacaaca gacgggcaca
    13381 cactacttga agcactcaag gcaagcttta ttgaggctta agcagtgggt tccctagtta
    13441 gccagagagc tcccaggctc agatctggtc taaccagaga gacccagtac aggcaaaacg
    13501 cgctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
    13561 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
    13621 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
    13681 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
    13741 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
    13801 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
    13861 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
    13921 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
    13981 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
    14041 ccgtaattga ttactattaa taactagtca ataatcaatg tcaacgcgta tatctggccc
    14101 gtacatcgcg aagcagcgca aaacgcctaa ccctaagcag attcttcatg caattgtcgg
    14161 tcaagccttg ccttgttgta gcttaaattt tgctcgcgca ctactcagcg acctccaaca
    14221 cacaagcagg gagcagatac tggcttaact atgcggcatc agagcagatt gtactgagag
    14281 tgcaccatag gggatcggga gatctcccga tccgtcgacg tcaggtggca cttttcgggg
    14341 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
    14401 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
    14461 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
    14521 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
    14581 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
    14641 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
    14701 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
    14761 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
    14821 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
    14881 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
    14941 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
    15001 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
    15061 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
    15121 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
    15181 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
    15241 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
    15301 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
    15361 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
    15421 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
    15481 ttc
//
