LOCUS       sequence-479442-       14921 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Expresses Lenti HBG site 1-ABE8e.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14921)
  AUTHORS   Xue N, Liu X, Zhang D, Wu Y, Zhong Y, Wang J, Fan W, Jiang H, Zhu B,
            Ge X, Gonzalez RVL, Chen L, Zhang S, She P, Zhong Z, Sun J, Chen X, 
            Wang L, Gu Z, Zhu P, Liu M, Li D, Zhong TP, Zhang X
  TITLE     Improving adenine and dual base editors through introduction of 
            TadA-8e and Rad51DBD.
  JOURNAL   Nat Commun. 2023 Mar 3;14(1):1224. doi: 10.1038/s41467-023-36887-1.
  PUBMED    36869044
REFERENCE   2  (bases 1 to 14921)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2023-03-3"; volume: "14(1):1224. doi"; pages: " 
            10.1038/s41467-023-36887-1"
COMMENT     Sequence Label: Lenti HBG site 1-ABE8e
FEATURES             Location/Qualifiers
     source          1..14921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1062..1295
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1480..1524
                     /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    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1984..2232
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1984..2004
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2155..2174
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2253..2328
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2397..2608
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2674..2694
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3289..3312,3313..3315,3316..7389)
                     /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             3289..7389
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7390..7437
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7453..7509
                     /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             join(7516..7518,7519..7521,7522..8232)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7516..7518,7519..7521,7522..8232)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7822..7841)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8169..8190
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        8268..8767
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8303..8326)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8700..8719
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             8780..9379
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8780..8799)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9276..9296
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9395..9983
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9448..9468)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9866..9877)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10055..10288
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10314..10331)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10320..10544
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10590..11018
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10677..10696)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10887..10908
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11027..11047)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11032..11361
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11212..11347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11274..11293
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11409..11456
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11475..11849
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    11979..12100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12016..12035)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12070..12089
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12149..12165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12149..12165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12162..12184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12173..12189
                     /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(12197..12203,12204..12221,12222..12227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12242..12263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12380..12397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12551..13139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12631..12650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13310..14101,14102..14170))
                     /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     13933..13952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14171..14275)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14350..14369)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        14541..14920
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taagagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
     2041 gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
     2101 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
     2161 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
     2221 ggacgaaaca ccgttgacca atagccttga cagttttaga gctagaaata gcaagttaaa
     2281 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttggtac
     2341 cgaattcgct agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc
     2401 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg
     2461 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     2521 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     2581 tcgcaacggg tttgccgcca gaacacaggg gcgcgcctct agagcgatcg cgccaccatg
     2641 aaacggacag ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtctctgag
     2701 gtggagtttt cccacgagta ctggatgaga catgccctga ccctggccaa gagggcacgg
     2761 gatgagaggg aggtgcctgt gggagccgtg ctggtgctga acaatagagt gatcggcgag
     2821 ggctggaaca gagccatcgg cctgcacgac ccaacagccc atgccgaaat tatggccctg
     2881 agacagggcg gcctggtcat gcagaactac agactgattg acgccaccct gtacgtgaca
     2941 ttcgagcctt gcgtgatgtg cgccggcgcc atgatccact ctaggatcgg ccgcgtggtg
     3001 tttggcgtga ggaactcaaa aagaggcgcc gcaggctccc tgatgaacgt gctgaactac
     3061 cccggcatga atcaccgcgt cgaaattacc gagggaatcc tggcagatga atgtgccgcc
     3121 ctgctgtgcg atttctatcg gatgcctaga caggtgttca atgctcagaa gaaggcccag
     3181 agctccatca actccggagg atctagcgga ggctcctctg gctctgagac acctggcaca
     3241 agcgagagcg caacacctga aagcagcggg ggcagcagcg gggggtcaga caagaagtac
     3301 agcatcggcc tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
     3361 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
     3421 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
     3481 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
     3541 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
     3601 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
     3661 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
     3721 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
     3781 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     3841 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
     3901 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     3961 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc
     4021 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     4081 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     4141 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     4201 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     4261 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     4321 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     4381 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     4441 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     4501 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     4561 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     4621 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     4681 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     4741 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     4801 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     4861 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     4921 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     4981 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     5041 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     5101 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     5161 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     5221 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     5281 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     5341 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     5401 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     5461 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     5521 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     5581 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     5641 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     5701 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     5761 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc
     5821 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
     5881 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     5941 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     6001 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     6061 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     6121 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     6181 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     6241 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     6301 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     6361 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     6421 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     6481 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     6541 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     6601 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
     6661 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
     6721 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
     6781 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     6841 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     6901 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
     6961 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     7021 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     7081 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     7141 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     7201 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     7261 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
     7321 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     7381 ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggga
     7441 tccggcagcg gcgcaacaaa cttctctctg ctgaaacaag ccggagatgt cgaagagaat
     7501 cctggaccgc tcgagatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     7561 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     7621 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     7681 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     7741 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     7801 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     7861 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     7921 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     7981 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     8041 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     8101 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     8161 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     8221 gagctgtaca agtactcaga tctggagctc aagtgagcta gcctaccggg taggggaggc
     8281 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
     8341 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
     8401 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
     8461 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
     8521 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
     8581 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
     8641 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
     8701 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
     8761 cctttcggaa ttcgccacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     8821 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     8881 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     8941 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     9001 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     9061 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     9121 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca
     9181 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     9241 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     9301 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     9361 ccgcaagccc ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
     9421 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     9481 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     9541 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     9601 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     9661 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     9721 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     9781 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     9841 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     9901 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     9961 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
    10021 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
    10081 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
    10141 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
    10201 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
    10261 ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac
    10321 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
    10381 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
    10441 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
    10501 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
    10561 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
    10621 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
    10681 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
    10741 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
    10801 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
    10861 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
    10921 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
    10981 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
    11041 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
    11101 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
    11161 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
    11221 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
    11281 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
    11341 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
    11401 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
    11461 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
    11521 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
    11581 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
    11641 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
    11701 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
    11761 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
    11821 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
    11881 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
    11941 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
    12001 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
    12061 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
    12121 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
    12181 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
    12241 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
    12301 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
    12361 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
    12421 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
    12481 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
    12541 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
    12601 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
    12661 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
    12721 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
    12781 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
    12841 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
    12901 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
    12961 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
    13021 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
    13081 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
    13141 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
    13201 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
    13261 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
    13321 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
    13381 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
    13441 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
    13501 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
    13561 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
    13621 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
    13681 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
    13741 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
    13801 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
    13861 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
    13921 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
    13981 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
    14041 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
    14101 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
    14161 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
    14221 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
    14281 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
    14341 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
    14401 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
    14461 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
    14521 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
    14581 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
    14641 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
    14701 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
    14761 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
    14821 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
    14881 ttggcagtac atctacgtat tagtcatcgc tattaccatg g
//
