LOCUS       sequence-479444-       15356 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Expresses Lenti HBG site 2-hyABE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15356)
  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 15356)
  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 2-hyABE
FEATURES             Location/Qualifiers
     source          1..15356
                     /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(3724..3747,3748..3750,3751..7824)
                     /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             3724..7824
                     /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             7825..7872
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7888..7944
                     /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(7951..7953,7954..7956,7957..8667)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7951..7953,7954..7956,7957..8667)
                     /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(8257..8276)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8604..8625
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        8703..9202
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8738..8761)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     9135..9154
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             9215..9814
                     /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(9215..9234)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9711..9731
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9830..10418
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9883..9903)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10301..10312)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10490..10723
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10749..10766)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10755..10979
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11025..11453
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11112..11131)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11322..11343
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11462..11482)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11467..11796
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11647..11782
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11709..11728
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11844..11891
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11910..12284
                     /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    12414..12535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12451..12470)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12505..12524
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12584..12600)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12584..12600)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12597..12619)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12608..12624
                     /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(12632..12638,12639..12656,12657..12662))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12677..12698
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12815..12832)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12986..13574)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13066..13085)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13745..14536,14537..14605))
                     /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     14368..14387
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14606..14710)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14785..14804)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        14976..15355
                     /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 ccgtggggaa ggggccccca aggttttaga 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 gggggtcagc aatgcagatg
     3301 cagcttgaag caaatgcaga tacttcagtg gaagaagaaa gctttggccc acaacccatt
     3361 tcacggttag agcagtgtgg cataaatgcc aacgatgtga agaaattgga agaagctgga
     3421 ttccatactg tggaggctgt tgcctatgcg ccaaagaagg agctaataaa tattaaggga
     3481 attagtgaag ccaaagctga taaaattctg gctgaggcag ctaaattagt tccaatgggt
     3541 ttcaccactg caactgaatt ccaccaaagg cggtcagaga tcatacagat tactactggc
     3601 tccaaagagc ttgacaaact acttcaatcc ggaggatcta gcggaggctc ctctggctct
     3661 gagacacctg gcacaagcga gagcgcaaca cctgaaagca gcgggggcag cagcgggggg
     3721 tcagacaaga agtacagcat cggcctggcc atcggcacca actctgtggg ctgggccgtg
     3781 atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg
     3841 cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag
     3901 gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc
     3961 tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga
     4021 ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc
     4081 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag
     4141 aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac
     4201 atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac
     4261 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc
     4321 atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga
     4381 cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac
     4441 ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag
     4501 gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc
     4561 cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc
     4621 ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct
     4681 atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg
     4741 cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc
     4801 ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg
     4861 gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg
     4921 aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac
     4981 gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc
     5041 gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc
     5101 agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa
     5161 gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag
     5221 aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg
     5281 tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg
     5341 agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc
     5401 gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc
     5461 tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt
     5521 atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg
     5581 ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc
     5641 cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc
     5701 aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg
     5761 gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac
     5821 agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg
     5881 cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca
     5941 gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg
     6001 atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga
     6061 atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc
     6121 gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg
     6181 gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat
     6241 atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc
     6301 gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag
     6361 aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg
     6421 accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag
     6481 ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac
     6541 actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc
     6601 aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac
     6661 taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag
     6721 taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag
     6781 atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc
     6841 aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg
     6901 cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt
     6961 gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg
     7021 cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc
     7081 gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc
     7141 tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg
     7201 aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac
     7261 tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag
     7321 tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg
     7381 cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc
     7441 cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa
     7501 cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg
     7561 atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag
     7621 cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc
     7681 cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa
     7741 gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc
     7801 gacctgtctc agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc cggccaggca
     7861 aaaaagaaaa agggatccgg cagcggcgca acaaacttct ctctgctgaa acaagccgga
     7921 gatgtcgaag agaatcctgg accgctcgag atggtgagca agggcgagga gctgttcacc
     7981 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     8041 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     8101 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     8161 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     8221 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     8281 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     8341 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     8401 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     8461 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     8521 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     8581 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     8641 actctcggca tggacgagct gtacaagtac tcagatctgg agctcaagtg agctagccta
     8701 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
     8761 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
     8821 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
     8881 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
     8941 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
     9001 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
     9061 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc
     9121 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
     9181 ttcctcatct ccgggccttt cggaattcgc caccatgacc gagtacaagc ccacggtgcg
     9241 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     9301 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     9361 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     9421 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     9481 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     9541 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     9601 agtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     9661 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     9721 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     9781 cacctggtgc atgacccgca agcccggtgc ctgaacgcgt taagtcgaca atcaacctct
     9841 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     9901 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     9961 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
    10021 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
    10081 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
    10141 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
    10201 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
    10261 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
    10321 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
    10381 tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact
    10441 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
    10501 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
    10561 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
    10621 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
    10681 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg
    10741 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
    10801 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
    10861 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
    10921 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
    10981 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
    11041 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
    11101 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
    11161 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
    11221 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
    11281 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
    11341 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
    11401 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
    11461 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
    11521 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
    11581 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
    11641 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
    11701 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
    11761 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
    11821 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
    11881 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
    11941 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
    12001 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
    12061 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
    12121 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
    12181 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
    12241 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt
    12301 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
    12361 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
    12421 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
    12481 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
    12541 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
    12601 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
    12661 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
    12721 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
    12781 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
    12841 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
    12901 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
    12961 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
    13021 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
    13081 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
    13141 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
    13201 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
    13261 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
    13321 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
    13381 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
    13441 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
    13501 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
    13561 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
    13621 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
    13681 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
    13741 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
    13801 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
    13861 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
    13921 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
    13981 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
    14041 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
    14101 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
    14161 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
    14221 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
    14281 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
    14341 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
    14401 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
    14461 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
    14521 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
    14581 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
    14641 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
    14701 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
    14761 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
    14821 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
    14881 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
    14941 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
    15001 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
    15061 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
    15121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
    15181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
    15241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
    15301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgg
//
