LOCUS       sequence-344625-       12685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Small-molecule controlled base editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12685)
  AUTHORS   Berrios KN, Evitt NH, DeWeerd RA, Ren D, Luo M, Barka A, Wang T, 
            Bartman CR, Lan Y, Green AM, Shi J, Kohli RM
  TITLE     Controllable genome editing with split-engineered base editors.
  JOURNAL   Nat Chem Biol. 2021 Dec;17(12):1262-1270. doi: 
            10.1038/s41589-021-00880-w. Epub 2021 Oct 18.
  PUBMED    34663942
REFERENCE   2  (bases 1 to 12685)
  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"; doi: 
            "10.1038/s41589-021-00880-w"; journalName: "Nat Chem Biol"; date: 
            "2021-12"; volume: "17"; issue: "12"; pages: "1262-1270"
COMMENT     Sequence Label: Lenti-evoA1-seBEC
FEATURES             Location/Qualifiers
     source          1..12685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     196..215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     449..466
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    583..604
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(619..624,625..642,643..649)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    657..673
                     /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     662..684
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     681..697
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     681..697
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     716..736
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        718..736
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        764..990
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             991..1171
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1218..1343
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1836..2069
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2254..2298
                     /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    2482..2599
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2668..2879
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2899..2908
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2908..2973
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(2974..2976,2977..3297)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             join(3892..3915,3916..3918,3919..7992)
                     /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             3892..7992
                     /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             8302..8550
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8593..8613
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8629..8685
                     /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             8686..9285
                     /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(8686..8705)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9182..9202
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9301..9889
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9354..9374)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9772..9783)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9961..10194
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10272..10393
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10309..10328)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10363..10382
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10433..10568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10588..10607)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10589..10607)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10617..10634)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10617..10633)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10626..10648)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10775..11230
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10862..10881)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11072..11093
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11256..11360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11361..11429,11430..12221)
                     /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(11579..11598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(12392..12685,1..295)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
       61 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      121 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      181 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      241 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      301 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      361 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
      421 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
      481 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      541 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
      601 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
      661 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
      721 taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata
      781 ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa
      841 aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg
      901 caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt
      961 gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc
     1021 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     1081 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     1141 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     1201 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     1261 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     1321 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     1381 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     1441 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     1501 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     1561 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     1621 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     1681 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     1741 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     1801 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     1861 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     1921 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     1981 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     2041 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     2101 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     2161 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     2221 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     2281 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     2341 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     2401 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     2461 ccgaggggac ccagaccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     2521 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     2581 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac tttggcgccg
     2641 gctcgagtgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     2701 agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact
     2761 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     2821 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
     2881 tgtcgtgacg cgggatccgc caccatggac tataaggacc acgacggaga ctacaaggat
     2941 catgatattg attacaaaga cgatgacgat aagatgggcg tacaagttga aactatcagc
     3001 cctggggacg gcagaacctt tccgaagagg ggacagacat gtgttgttca ctatactgga
     3061 atgttggaag atggtaagaa gttcgatagc agcagagata ggaataaacc atttaaattc
     3121 atgcttggca agcaagaagt gattaggggt tgggaagaag gtgtcgctca aatgagtgta
     3181 ggtcagaggg ctaagttaac aattagtcct gattatgctt atggcgctac aggtcatcca
     3241 ggaatcattc ccccacatgc tactcttgtt ttcgacgttg aattgcttaa gcttgaagga
     3301 tcaggttctg gatctggttc aggatcaggc tcacccgggc ttaataccag atgtagcatc
     3361 acatggtttc tgagctggtc cccttgcgga gagtgtagca gggccatcac cgagttcctg
     3421 tccagatatc caaatgtgac actgtttatc tacatcgcca ggctgtatca cctggcaaac
     3481 ccaaggaata ggcagggcct gcgcgatctg atcagctccg gcgtgaccat ccagatcatg
     3541 acagagcagg agtccggcta ctgctggcac aacttcgtga attattctcc tagcaacgag
     3601 tcccactggc ctaggtaccc acacctgtgg gtgcgcctgt acgtgctgga gctgtattgc
     3661 atcatcctgg gcctgccccc ttgtctgaat atcctgcgga gaaagcagag ccagctgacc
     3721 tcctttacaa tcgccctgca gtcttgtcac tatcagaggc tgccacccca catcctgtgg
     3781 gccacaggcc tgaagtctgg cggatctagc ggaggatcct ctggcagcga gacacccggg
     3841 acaagcgagt cagcaacacc agagagcagt ggcggcagca gcggcggcag cgacaagaag
     3901 tacagcatcg gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag
     3961 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
     4021 aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg
     4081 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
     4141 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
     4201 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
     4261 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
     4321 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
     4381 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
     4441 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
     4501 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     4561 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg
     4621 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     4681 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     4741 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     4801 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
     4861 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     4921 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac
     4981 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     5041 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     5101 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     5161 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     5221 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     5281 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     5341 ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     5401 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg
     5461 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     5521 aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     5581 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
     5641 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
     5701 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
     5761 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
     5821 gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg
     5881 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
     5941 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
     6001 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
     6061 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
     6121 gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc
     6181 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
     6241 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
     6301 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
     6361 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag
     6421 agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg
     6481 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg
     6541 cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag
     6601 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
     6661 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
     6721 gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
     6781 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
     6841 cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
     6901 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
     6961 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
     7021 tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag
     7081 acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg
     7141 aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc
     7201 ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag
     7261 gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg
     7321 gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg
     7381 gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc
     7441 aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc
     7501 gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac
     7561 gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag
     7621 ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac
     7681 tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac
     7741 gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag
     7801 caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc
     7861 aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac
     7921 gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag
     7981 ctgggaggtg acagcggcgg gagcggcggg agcgagggga gcactaatct gagcgacatc
     8041 attgagaaga agactgggaa acagctggtc attcaggagt ccatcctgat gctgcctgag
     8101 gaggtggagg aagtgatcgg caacaagcca gagtctgaca tcctggtgca caccgcctac
     8161 gacgagtcca cagatgagaa tgtgatgctg ctgacctctg acgcccccga gtataagcct
     8221 tgggccctgg tcatccagga ttctaacggc gagaataaga tcaagatgct gagcggagga
     8281 tccggaggat ctggaggcag caccaacctg tctgacatca tcgagaagga gacaggcaag
     8341 cagctggtca tccaggagag catcctgatg ctgcccgaag aagtcgaaga agtgatcgga
     8401 aacaagcctg agagcgatat cctggtccat accgcctacg acgagagtac cgacgaaaat
     8461 gtgatgctgc tgacatccga cgccccagag tataagccct gggctctggt catccaggat
     8521 tccaacggag agaacaaaat caaaatgctg tctggcggct caaaaagaac cgccgacggc
     8581 agcgaattcg agcccaagaa gaagaggaaa gtcgctagcg gcagcggcgc caccaacttc
     8641 agcctgctga agcaggccgg cgacgtggag gagaaccccg gccccatgac cgagtacaag
     8701 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     8761 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     8821 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     8881 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     8941 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     9001 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     9061 gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     9121 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     9181 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     9241 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaacgcg ttaagtcgac
     9301 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     9361 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     9421 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     9481 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     9541 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     9601 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     9661 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     9721 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     9781 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     9841 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa
     9901 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     9961 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
    10021 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
    10081 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
    10141 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtacgt
    10201 atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
    10261 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
    10321 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
    10381 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
    10441 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
    10501 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
    10561 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
    10621 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
    10681 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
    10741 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
    10801 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
    10861 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
    10921 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
    10981 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
    11041 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
    11101 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
    11161 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
    11221 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
    11281 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
    11341 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
    11401 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
    11461 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
    11521 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
    11581 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
    11641 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
    11701 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
    11761 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
    11821 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
    11881 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
    11941 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
    12001 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
    12061 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
    12121 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
    12181 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
    12241 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
    12301 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
    12361 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
    12421 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
    12481 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
    12541 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
    12601 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
    12661 ggttggactc aagacgatag ttacc
//
